2024-03-28T15:34:51Zhttp://repository.helmholtz-hzi.de/oai/requestoai:repository.helmholtz-hzi.de:10033/784942019-08-30T11:30:58Zcom_10033_620589col_10033_620590
Haid, Sibylle
Pietschmann, Thomas
Pécheur, Eve-Isabelle
Department for Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research, a Joint Venture of Hannover Medical School and the Helmholtz-Centre for Infection Research, Hannover 30625, Germany.
2009-08-25T13:22:55Z
2009-08-25T13:22:55Z
2009-06-26
Low pH-dependent hepatitis C virus membrane fusion depends on E2 integrity, target lipid composition, and density of virus particles. 2009, 284 (26):17657-67 J. Biol. Chem.
0021-9258
19411248
10.1074/jbc.M109.014647
http://hdl.handle.net/10033/78494
The Journal of biological chemistry
Hepatitis C virus (HCV) is an enveloped, positive strand RNA virus of about 9.6 kb. Like all enveloped viruses, the HCV membrane fuses with the host cell membrane during the entry process and thereby releases the genome into the cytoplasm, initiating the viral replication cycle. To investigate the features of HCV membrane fusion, we developed an in vitro fusion assay using cell culture-produced HCV and fluorescently labeled liposomes. With this model we could show that HCV-mediated fusion can be triggered in a receptor-independent but pH-dependent manner and that fusion of the HCV particles with liposomes is dependent on the viral dose and on the lipid composition of the target membranes. In addition CBH-5, an HCV E2-specific antibody, inhibited fusion in a dose-dependent manner. Interestingly, point mutations in E2, known to abrogate HCV glycoprotein-mediated fusion in a cell-based assay, altered or even abolished fusion in the liposome-based assay. When assaying the fusion properties of HCV particles with different buoyant density, we noted higher fusogenicity of particles with lower density. This could be attributable to inherently different properties of low density particles, to association of these particles with factors stimulating fusion, or to co-flotation of factors enhancing fusion activity in trans. Taken together, these data show the important role of lipids of both the viral and target membranes in HCV-mediated fusion, point to a crucial role played by the E2 glycoprotein in the process of HCV fusion, and reveal an important behavior of HCV of different densities with regard to fusion.
en
Carcinoma, Hepatocellular
Cholesterol
Electroporation
Hepacivirus
Hepatitis C
Hepatitis C Antibodies
Humans
Hydrogen-Ion Concentration
Immunoenzyme Techniques
Immunoprecipitation
Indoles
Liposomes
Liver Neoplasms
Luciferases
RNA, Viral
Sphingomyelins
Transcription, Genetic
Tumor Cells, Cultured
Viral Envelope Proteins
Virion
Virus Internalization
Low pH-dependent hepatitis C virus membrane fusion depends on E2 integrity, target lipid composition, and density of virus particles.
Article2018-06-12T23:04:16ZHepatitis C virus (HCV) is an enveloped, positive strand RNA virus of about 9.6 kb. Like all enveloped viruses, the HCV membrane fuses with the host cell membrane during the entry process and thereby releases the genome into the cytoplasm, initiating the viral replication cycle. To investigate the features of HCV membrane fusion, we developed an in vitro fusion assay using cell culture-produced HCV and fluorescently labeled liposomes. With this model we could show that HCV-mediated fusion can be triggered in a receptor-independent but pH-dependent manner and that fusion of the HCV particles with liposomes is dependent on the viral dose and on the lipid composition of the target membranes. In addition CBH-5, an HCV E2-specific antibody, inhibited fusion in a dose-dependent manner. Interestingly, point mutations in E2, known to abrogate HCV glycoprotein-mediated fusion in a cell-based assay, altered or even abolished fusion in the liposome-based assay. When assaying the fusion properties of HCV particles with different buoyant density, we noted higher fusogenicity of particles with lower density. This could be attributable to inherently different properties of low density particles, to association of these particles with factors stimulating fusion, or to co-flotation of factors enhancing fusion activity in trans. Taken together, these data show the important role of lipids of both the viral and target membranes in HCV-mediated fusion, point to a crucial role played by the E2 glycoprotein in the process of HCV fusion, and reveal an important behavior of HCV of different densities with regard to fusion.oai:repository.helmholtz-hzi.de:10033/1076192019-08-30T11:30:32Zcom_10033_620589col_10033_620590
Ciesek, Sandra
Steinmann, Eike
Iken, Markus
Ott, Michael
Helfritz, Fabian A
Wappler, Ilka
Manns, Michael P
Wedemeyer, Heiner
Pietschmann, Thomas
Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Hannover, Germany.
2010-07-14T09:43:57Z
2010-07-14T09:43:57Z
2010-05
Glucocorticosteroids increase cell entry by hepatitis C virus. 2010, 138 (5):1875-84 Gastroenterology
1528-0012
20152835
10.1053/j.gastro.2010.02.004
http://hdl.handle.net/10033/107619
Gastroenterology
BACKGROUND & AIMS: Corticosteroids are used as immunosuppressants in patients with autoimmune disorders and transplant recipients. However, these drugs worsen hepatitis C virus (HCV) recurrence after liver transplantation, suggesting that they may directly exacerbate HCV infection. METHODS: The influence of immunosuppressive drugs on HCV replication, assembly, and entry was assessed in Huh-7.5 cells and primary human hepatocytes using cell culture- and patient-derived HCV. Replication was quantified by immunofluorescence, luciferase assays, quantitative reverse-transcriptase polymerase chain reaction, or core enzyme-linked immunosorbent assays. Expression of HCV entry factors was evaluated by cell sorting and immunoblot analyses. RESULTS: Glucocorticosteroids slightly reduced HCV RNA replication but increased efficiency of HCV entry by up to 10-fold. This was independent of HCV genotype but specific to HCV because vesicular stomatitis virus glycoprotein-dependent infection was not affected by these drugs. The increase in HCV entry was accompanied by up-regulation of messenger RNA and protein levels of occludin and the scavenger receptor class B type I-2 host cell proteins required for HCV infection; increase of entry by glucocorticosteroids was ablated by RU-486, an inhibitor of glucocorticosteroid signaling. Glucocorticosteroids increased propagation of cell culture-derived HCV approximately 5- to 10-fold in partially differentiated human hepatoma cells and increased infection of primary human hepatocytes by cell culture- and patient-derived HCV. CONCLUSIONS: Glucocorticosteroides specifically increase HCV entry by up-regulating the cell entry factors occludin and scavenger receptor class B type I. Our data suggest that the potential effects of high-dose glucocorticosteroids on HCV infection in vivo may be due to increased HCV dissemination.
en
Cells, Cultured
Dose-Response Relationship, Drug
Genotype
Glucocorticoids
Hepacivirus
Hepatocytes
Hormone Antagonists
Humans
Immunosuppressive Agents
Membrane Proteins
Mifepristone
Prednisolone
RNA, Messenger
RNA, Viral
Scavenger Receptors, Class B
Time Factors
Virus Internalization
Virus Replication
Glucocorticosteroids increase cell entry by hepatitis C virus.
Article2018-06-13T19:30:17ZBACKGROUND & AIMS: Corticosteroids are used as immunosuppressants in patients with autoimmune disorders and transplant recipients. However, these drugs worsen hepatitis C virus (HCV) recurrence after liver transplantation, suggesting that they may directly exacerbate HCV infection. METHODS: The influence of immunosuppressive drugs on HCV replication, assembly, and entry was assessed in Huh-7.5 cells and primary human hepatocytes using cell culture- and patient-derived HCV. Replication was quantified by immunofluorescence, luciferase assays, quantitative reverse-transcriptase polymerase chain reaction, or core enzyme-linked immunosorbent assays. Expression of HCV entry factors was evaluated by cell sorting and immunoblot analyses. RESULTS: Glucocorticosteroids slightly reduced HCV RNA replication but increased efficiency of HCV entry by up to 10-fold. This was independent of HCV genotype but specific to HCV because vesicular stomatitis virus glycoprotein-dependent infection was not affected by these drugs. The increase in HCV entry was accompanied by up-regulation of messenger RNA and protein levels of occludin and the scavenger receptor class B type I-2 host cell proteins required for HCV infection; increase of entry by glucocorticosteroids was ablated by RU-486, an inhibitor of glucocorticosteroid signaling. Glucocorticosteroids increased propagation of cell culture-derived HCV approximately 5- to 10-fold in partially differentiated human hepatoma cells and increased infection of primary human hepatocytes by cell culture- and patient-derived HCV. CONCLUSIONS: Glucocorticosteroides specifically increase HCV entry by up-regulating the cell entry factors occludin and scavenger receptor class B type I. Our data suggest that the potential effects of high-dose glucocorticosteroids on HCV infection in vivo may be due to increased HCV dissemination.oai:repository.helmholtz-hzi.de:10033/1181942019-08-30T11:30:58Zcom_10033_620589col_10033_620590
Ciesek, Sandra
Friesland, Martina
Steinmann, Jörg
Becker, Britta
Wedemeyer, Heiner
Manns, Michael P
Steinmann, Jochen
Pietschmann, Thomas
Steinmann, Eike
Division of Experimental Virology, Twincore, Centre for Experimental and Clinical Infection Research, Joint venture between Hannover Medical School, Hannover, Germany.
2010-12-21T15:25:11Z
2010-12-21T15:25:11Z
2010-06-15
How stable is the hepatitis C virus (HCV)? Environmental stability of HCV and its susceptibility to chemical biocides. 2010, 201 (12):1859-66 J. Infect. Dis.
1537-6613
20441517
10.1086/652803
http://hdl.handle.net/10033/118194
The Journal of infectious diseases
In the absence of a cell culture system for propagation of the hepatitis C virus (HCV), the antiviral activity of disinfectants against HCV was extrapolated from studies with the bovine viral diarrhea virus. The recent development of an HCV infection system allowed the direct assessment of environmental stability and susceptibility to chemical disinfectants.
en
Cell Line
Disinfectants
Hepacivirus
Humans
Microbial Viability
RNA, Viral
Reverse Transcriptase Polymerase Chain Reaction
Temperature
Time Factors
Transfection
Virus Inactivation
How stable is the hepatitis C virus (HCV)? Environmental stability of HCV and its susceptibility to chemical biocides.
Article2018-06-13T01:32:45ZIn the absence of a cell culture system for propagation of the hepatitis C virus (HCV), the antiviral activity of disinfectants against HCV was extrapolated from studies with the bovine viral diarrhea virus. The recent development of an HCV infection system allowed the direct assessment of environmental stability and susceptibility to chemical disinfectants.oai:repository.helmholtz-hzi.de:10033/1463552019-08-30T11:27:46Zcom_10033_620589col_10033_620590
Frentzen, Anne
Hüging, Kathrin
Bitzegeio, Julia
Friesland, Martina
Haid, Sibylle
Gentzsch, Juliane
Hoffmann, Markus
Lindemann, Dirk
Zimmer, Gert
Zielecki, Florian
Weber, Friedemann
Steinmann, Eike
Pietschmann, Thomas
Division of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research, Hannover, Germany.
2011-10-21T13:45:31Z
2011-10-21T13:45:31Z
2011-04
Completion of hepatitis C virus replication cycle in heterokaryons excludes dominant restrictions in human non-liver and mouse liver cell lines. 2011, 7 (4):e1002029 PLoS Pathog.
1553-7374
21552323
10.1371/journal.ppat.1002029
http://hdl.handle.net/10033/146355
PLoS pathogens
Hepatitis C virus (HCV) is hepatotropic and only infects humans and chimpanzees. Consequently, an immunocompetent small animal model is lacking. The restricted tropism of HCV likely reflects specific host factor requirements. We investigated if dominant restriction factors expressed in non-liver or non-human cell lines inhibit HCV propagation thus rendering these cells non-permissive. To this end we explored if HCV completes its replication cycle in heterokaryons between human liver cell lines and non-permissive cell lines from human non-liver or mouse liver origin. Despite functional viral pattern recognition pathways and responsiveness to interferon, virus production was observed in all fused cells and was only ablated when cells were treated with exogenous interferon. These results exclude that constitutive or virus-induced expression of dominant restriction factors prevents propagation of HCV in these cell types, which has important implications for HCV tissue and species tropism. In turn, these data strongly advocate transgenic approaches of crucial human HCV cofactors to establish an immunocompetent small animal model.
en
Animals
Cell Fusion
Cell Line
HEK293 Cells
Hela Cells
Hepacivirus
Humans
Interferon-alpha
Mice
Models, Animal
Transfection
Virus Replication
Completion of hepatitis C virus replication cycle in heterokaryons excludes dominant restrictions in human non-liver and mouse liver cell lines.
Article2018-06-13T00:00:37ZHepatitis C virus (HCV) is hepatotropic and only infects humans and chimpanzees. Consequently, an immunocompetent small animal model is lacking. The restricted tropism of HCV likely reflects specific host factor requirements. We investigated if dominant restriction factors expressed in non-liver or non-human cell lines inhibit HCV propagation thus rendering these cells non-permissive. To this end we explored if HCV completes its replication cycle in heterokaryons between human liver cell lines and non-permissive cell lines from human non-liver or mouse liver origin. Despite functional viral pattern recognition pathways and responsiveness to interferon, virus production was observed in all fused cells and was only ablated when cells were treated with exogenous interferon. These results exclude that constitutive or virus-induced expression of dominant restriction factors prevents propagation of HCV in these cell types, which has important implications for HCV tissue and species tropism. In turn, these data strongly advocate transgenic approaches of crucial human HCV cofactors to establish an immunocompetent small animal model.oai:repository.helmholtz-hzi.de:10033/2136952019-08-30T11:37:23Zcom_10033_620589col_10033_620590
von Hahn, Thomas
Schulze, Andreas
Chicano Wust, Ivan
Heidrich, Benjamin
Becker, Thomas
Steinmann, Eike
Helfritz, Fabian A
Rohrmann, Katrin
Urban, Stephan
Manns, Michael P
Pietschmann, Thomas
Ciesek, Sandra
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
2012-03-01T10:28:46Z
2012-03-01T10:28:46Z
2011
The novel immunosuppressive protein kinase C inhibitor sotrastaurin has no pro-viral effects on the replication cycle of hepatitis B or C virus. 2011, 6 (9):e24142 PLoS ONE
1932-6203
21909416
10.1371/journal.pone.0024142
http://hdl.handle.net/10033/213695
PloS one
The pan-protein kinase C (PKC) inhibitor sotrastaurin (AEB071) is a novel immunosuppressant currently in phase II trials for immunosuppression after solid organ transplantation. Besides T-cell activation, PKC affects numerous cellular processes that are potentially important for the replication of hepatitis B virus (HBV) and hepatitis C virus (HCV), major blood-borne pathogens prevalent in solid organ transplant recipients. This study uses state of the art virological assays to assess the direct, non-immune mediated effects of sotrastaurin on HBV and HCV. Most importantly, sotrastaurin had no pro-viral effect on either HBV or HCV. In the presence of high concentrations of sotrastaurin, well above those used clinically and close to levels where cytotoxic effects become detectable, there was a reduction of HCV and HBV replication. This reduction is very likely due to cytotoxic and/or anti-proliferative effects rather than direct anti-viral activity of the drug. Replication cycle stages other than genome replication such as viral cell entry and spread of HCV infection directly between adjacent cells was clearly unaffected by sotrastaurin. These data support the evaluation of sotrastaurin in HBV and/or HCV infected transplant recipients.
en
Calcineurin
Cell Line, Tumor
Cyclosporine
Hepacivirus
Hepatitis B virus
Humans
Immunosuppressive Agents
Protein Kinase C
Protein Kinase Inhibitors
Pyrroles
Quinazolines
RNA, Viral
Virus Internalization
Virus Replication
The novel immunosuppressive protein kinase C inhibitor sotrastaurin has no pro-viral effects on the replication cycle of hepatitis B or C virus.
Article2018-06-13T09:21:19ZThe pan-protein kinase C (PKC) inhibitor sotrastaurin (AEB071) is a novel immunosuppressant currently in phase II trials for immunosuppression after solid organ transplantation. Besides T-cell activation, PKC affects numerous cellular processes that are potentially important for the replication of hepatitis B virus (HBV) and hepatitis C virus (HCV), major blood-borne pathogens prevalent in solid organ transplant recipients. This study uses state of the art virological assays to assess the direct, non-immune mediated effects of sotrastaurin on HBV and HCV. Most importantly, sotrastaurin had no pro-viral effect on either HBV or HCV. In the presence of high concentrations of sotrastaurin, well above those used clinically and close to levels where cytotoxic effects become detectable, there was a reduction of HCV and HBV replication. This reduction is very likely due to cytotoxic and/or anti-proliferative effects rather than direct anti-viral activity of the drug. Replication cycle stages other than genome replication such as viral cell entry and spread of HCV infection directly between adjacent cells was clearly unaffected by sotrastaurin. These data support the evaluation of sotrastaurin in HBV and/or HCV infected transplant recipients.oai:repository.helmholtz-hzi.de:10033/2139892019-08-30T11:30:58Zcom_10033_620589col_10033_620590
Steinmann, J
Kaase, M
Gatermann, S
Popp, W
Steinmann, E
Damman, M
Paul, A
Saner, F
Buer, J
Rath, Pm
Institute of Medical Microbiology, University Hospital Essen, University of Duisburg- Essen, Essen, Germany. joerg.steinmann@uk-essen.de
2012-03-02T10:50:33Z
2012-03-02T10:50:33Z
2011
Outbreak due to a Klebsiella pneumoniae strain harbouring KPC-2 and VIM-1 in a German university hospital, July 2010 to January 2011. 2011, 16 (33): Euro Surveill.
1560-7917
21871227
http://hdl.handle.net/10033/213989
Euro surveillance : bulletin européen sur les maladies transmissibles = European communicable disease bulletin
We describe the epidemiology and characteristics of the pathogen and patients (n=7) associated with an outbreak of a carbapenem-resistant Klebsiella pneumoniae (CRKP) strain in a German university hospital from July 2010 to January 2011. Species identification and detection of carbapenem resistance were carried out using standard microbiological procedures. Carbapenemases were detected by phenotypic methods and specific polymerase chain reactions (PCRs). DNA fingerprinting profiles were performed with repetitive sequence-based PCR. Medical records of colonised or infected patients were retrospectively reviewed. Antibiotic resistance profiles, PCR-specific amplification products and genotyping demonstrated that the outbreak occurred because of the spread of a single CRKP clone harbouring both KPC-2 and VIM-1. Five of the seven patients had invasive infections with the CRKP strain; the deaths of four of them were directly related to the infection. Early implementation of infection control interventions brought about efficient containment of further cross-transmission. Rapid dissemination of carbapenemase-producing Enterobacteriaceae is a serious concern in patient care and is a problem that has emerged in western Europe.
en
Adult
Aged
Anti-Bacterial Agents
Carbenicillin
Cross Infection
DNA Fingerprinting
Disease Outbreaks
Drug Resistance, Multiple, Bacterial
Female
Genotype
Germany
Hospitals, University
Humans
Intensive Care Units
Klebsiella Infections
Klebsiella pneumoniae
Male
Microbial Sensitivity Tests
Middle Aged
Polymerase Chain Reaction
Retrospective Studies
Sequence Analysis, DNA
Young Adult
beta-Lactamases
Outbreak due to a Klebsiella pneumoniae strain harbouring KPC-2 and VIM-1 in a German university hospital, July 2010 to January 2011.
Article2018-06-13T01:15:55ZWe describe the epidemiology and characteristics of the pathogen and patients (n=7) associated with an outbreak of a carbapenem-resistant Klebsiella pneumoniae (CRKP) strain in a German university hospital from July 2010 to January 2011. Species identification and detection of carbapenem resistance were carried out using standard microbiological procedures. Carbapenemases were detected by phenotypic methods and specific polymerase chain reactions (PCRs). DNA fingerprinting profiles were performed with repetitive sequence-based PCR. Medical records of colonised or infected patients were retrospectively reviewed. Antibiotic resistance profiles, PCR-specific amplification products and genotyping demonstrated that the outbreak occurred because of the spread of a single CRKP clone harbouring both KPC-2 and VIM-1. Five of the seven patients had invasive infections with the CRKP strain; the deaths of four of them were directly related to the infection. Early implementation of infection control interventions brought about efficient containment of further cross-transmission. Rapid dissemination of carbapenemase-producing Enterobacteriaceae is a serious concern in patient care and is a problem that has emerged in western Europe.oai:repository.helmholtz-hzi.de:10033/2164742019-08-30T11:32:41Zcom_10033_620589col_10033_620590
Doerrbecker, Juliane
Friesland, Martina
Ciesek, Sandra
Erichsen, Thomas J
Mateu-Gelabert, Pedro
Steinmann, Jörg
Steinmann, Jochen
Pietschmann, Thomas
Steinmann, Eike
Division of Experimental Virology, Twincore, Centre for Experimental and Clinical, Infection Research, a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Germany.
2012-03-23T14:33:51Z
2012-03-23T14:33:51Z
2011-12-15
Inactivation and survival of hepatitis C virus on inanimate surfaces. 2011, 204 (12):1830-8 J. Infect. Dis.
1537-6613
22013220
10.1093/infdis/jir535
http://hdl.handle.net/10033/216474
The Journal of infectious diseases
Hepatitis C virus (HCV) cross-contamination from inanimate surfaces or objects has been implicated in transmission of HCV in health-care settings and among injection drug users. We established HCV-based carrier and drug transmission assays that simulate practical conditions to study inactivation and survival of HCV on inanimate surfaces.
en
Archived with thanks to The Journal of infectious diseases
1-Propanol
2-Propanol
Disinfectants
Equipment Contamination
Ethanol
Glutaral
Hepacivirus
Hepatitis C
Humans
Microbial Viability
Peroxides
Quaternary Ammonium Compounds
Substance Abuse, Intravenous
Temperature
Time Factors
Virus Inactivation
Inactivation and survival of hepatitis C virus on inanimate surfaces.
Article2012-12-15T00:00:00ZHepatitis C virus (HCV) cross-contamination from inanimate surfaces or objects has been implicated in transmission of HCV in health-care settings and among injection drug users. We established HCV-based carrier and drug transmission assays that simulate practical conditions to study inactivation and survival of HCV on inanimate surfaces.oai:repository.helmholtz-hzi.de:10033/2229982019-08-30T11:24:31Zcom_10033_620589col_10033_620590
Chhatwal, Patrick
Bankwitz, Dorothea
Gentzsch, Juliane
Frentzen, Anne
Schult, Philipp
Lohmann, Volker
Pietschmann, Thomas
Department of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany.
2012-05-10T09:24:05Z
2012-05-10T09:24:05Z
2012
Bile Acids Specifically Increase Hepatitis C Virus RNA-Replication. 2012, 7 (4):e36029 PLoS ONE
1932-6203
22558311
10.1371/journal.pone.0036029
http://hdl.handle.net/10033/222998
PloS one
Hepatitis C virus (HCV) patients with high serum levels of bile acids (BAs) respond poorly to IFN therapy. BAs have been shown to increase RNA-replication of genotype 1 but not genotype 2a replicons. Since BAs modulate lipid metabolism including lipoprotein secretion and as HCV depends on lipids and lipoproteins during RNA-replication, virus production and cell entry, BAs may affect multiple steps of the HCV life cycle. Therefore, we analyzed the influence of BAs on individual steps of virus replication.
en
Archived with thanks to PloS one
Bile Acids Specifically Increase Hepatitis C Virus RNA-Replication.
Article2018-06-13T09:15:30ZHepatitis C virus (HCV) patients with high serum levels of bile acids (BAs) respond poorly to IFN therapy. BAs have been shown to increase RNA-replication of genotype 1 but not genotype 2a replicons. Since BAs modulate lipid metabolism including lipoprotein secretion and as HCV depends on lipids and lipoproteins during RNA-replication, virus production and cell entry, BAs may affect multiple steps of the HCV life cycle. Therefore, we analyzed the influence of BAs on individual steps of virus replication.oai:repository.helmholtz-hzi.de:10033/2256022019-08-30T11:27:46Zcom_10033_620589col_10033_620590
Haid, Sibylle
Novodomská, Alexandra
Gentzsch, Juliane
Grethe, Christina
Geuenich, Silvia
Bankwitz, Dorothea
Chhatwal, Patrick
Jannack, Beate
Hennebelle, Thierry
Bailleul, Francois
Keppler, Oliver T
Pönisch, Marion
Bartenschlager, Ralf
Hernandez, Céline
Lemasson, Matthieu
Rosenberg, Arielle
Wong-Staal, Flossie
Davioud-Charvet, Elisabeth
Pietschmann, Thomas
Division of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany.
2012-05-23T14:24:09Z
2012-05-23T14:24:09Z
2012-03-27
A Plant-Derived Flavonoid Inhibits Entry of All HCV Genotypes Into Human Hepatocytes. 2012:notGastroenterology
1528-0012
22465429
10.1053/j.gastro.2012.03.036
http://hdl.handle.net/10033/225602
Gastroenterology
BACKGROUND & AIMS: Interferon-based therapies for hepatitis C virus (HCV) infection are limited by side effects and incomplete response rates, particularly among transplant recipients. We screened a library of plant-derived small molecules to identify HCV inhibitors with novel mechanisms. METHODS: We isolated phenolic compounds from Marrubium peregrinum L (Lamiaceae). Replication of HCV RNA, virus production, and cell entry were monitored using replicons and infectious HCV. Inhibition of HCV was measured in hepatoma cells and primary human hepatocytes using luciferase reporter gene assays, core enzyme-linked immunosorbent assays, or infectivity titration. We tested the bioavailability of the compound in mice. RESULTS: We identified a flavonoid, ladanein (BJ486K), with unreported antiviral activity and established its oral bioavailability in mice. Natural and synthetic BJ486K inhibited a post-attachment entry step, but not RNA replication or assembly; its inhibitory concentration 50% was 2.5 μm. BJ486K was effective against all major HCV genotypes, including a variant that is resistant to an entry inhibitor; it prevented infection of primary human hepatocytes. Combined administration of BJ486K and cyclosporine A had a synergistic effect in inhibition of HCV infection. CONCLUSIONS: BJ486K has oral bioavailability and interferes with entry of HCV into cultured human hepatocytes. It synergizes with cyclosporine A to inhibit HCV infection. Its inhibitory effects are independent of HCV genotype, including a variant that is resistant to an entry inhibitor against scavenger receptor class B type I. Flavonoid derivatives therefore might be developed as components of combination therapies because they are potent, broadly active, inhibitors of HCV entry that could prevent graft reinfection after liver transplantation.
ENG
Archived with thanks to Gastroenterology
A Plant-Derived Flavonoid Inhibits Entry of All HCV Genotypes Into Human Hepatocytes.
Article2018-06-13T01:09:53ZBACKGROUND & AIMS: Interferon-based therapies for hepatitis C virus (HCV) infection are limited by side effects and incomplete response rates, particularly among transplant recipients. We screened a library of plant-derived small molecules to identify HCV inhibitors with novel mechanisms. METHODS: We isolated phenolic compounds from Marrubium peregrinum L (Lamiaceae). Replication of HCV RNA, virus production, and cell entry were monitored using replicons and infectious HCV. Inhibition of HCV was measured in hepatoma cells and primary human hepatocytes using luciferase reporter gene assays, core enzyme-linked immunosorbent assays, or infectivity titration. We tested the bioavailability of the compound in mice. RESULTS: We identified a flavonoid, ladanein (BJ486K), with unreported antiviral activity and established its oral bioavailability in mice. Natural and synthetic BJ486K inhibited a post-attachment entry step, but not RNA replication or assembly; its inhibitory concentration 50% was 2.5 μm. BJ486K was effective against all major HCV genotypes, including a variant that is resistant to an entry inhibitor; it prevented infection of primary human hepatocytes. Combined administration of BJ486K and cyclosporine A had a synergistic effect in inhibition of HCV infection. CONCLUSIONS: BJ486K has oral bioavailability and interferes with entry of HCV into cultured human hepatocytes. It synergizes with cyclosporine A to inhibit HCV infection. Its inhibitory effects are independent of HCV genotype, including a variant that is resistant to an entry inhibitor against scavenger receptor class B type I. Flavonoid derivatives therefore might be developed as components of combination therapies because they are potent, broadly active, inhibitors of HCV entry that could prevent graft reinfection after liver transplantation.oai:repository.helmholtz-hzi.de:10033/2470122019-08-30T11:24:31Zcom_10033_620589col_10033_620590
Menzel, Nicolas
Fischl, Wolfgang
Hueging, Kathrin
Bankwitz, Dorothea
Frentzen, Anne
Haid, Sibylle
Gentzsch, Juliane
Kaderali, Lars
Bartenschlager, Ralf
Pietschmann, Thomas
Division of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Hannover, Germany.
2012-10-04T14:40:26Z
2012-10-04T14:40:26Z
2012-07
MAP-Kinase Regulated Cytosolic Phospholipase A2 Activity Is Essential for Production of Infectious Hepatitis C Virus Particles. 2012, 8 (7):e1002829 PLoS Pathog.
1553-7374
22911431
10.1371/journal.ppat.1002829
http://hdl.handle.net/10033/247012
PLoS pathogens
Hepatitis C virus (HCV) has infected around 160 million individuals. Current therapies have limited efficacy and are fraught with side effects. To identify cellular HCV dependency factors, possible therapeutic targets, we manipulated signaling cascades with pathway-specific inhibitors. Using this approach we identified the MAPK/ERK regulated, cytosolic, calcium-dependent, group IVA phospholipase A2 (PLA2G4A) as a novel HCV dependency factor. Inhibition of PLA2G4A activity reduced core protein abundance at lipid droplets, core envelopment and secretion of particles. Moreover, released particles displayed aberrant protein composition and were 100-fold less infectious. Exogenous addition of arachidonic acid, the cleavage product of PLA2G4A-catalyzed lipolysis, but not other related poly-unsaturated fatty acids restored infectivity. Strikingly, production of infectious Dengue virus, a relative of HCV, was also dependent on PLA2G4A. These results highlight previously unrecognized parallels in the assembly pathways of these human pathogens, and define PLA2G4A-dependent lipolysis as crucial prerequisite for production of highly infectious viral progeny.
en
Archived with thanks to PLoS pathogens
MAP-Kinase Regulated Cytosolic Phospholipase A2 Activity Is Essential for Production of Infectious Hepatitis C Virus Particles.
Article2018-06-12T23:27:28ZHepatitis C virus (HCV) has infected around 160 million individuals. Current therapies have limited efficacy and are fraught with side effects. To identify cellular HCV dependency factors, possible therapeutic targets, we manipulated signaling cascades with pathway-specific inhibitors. Using this approach we identified the MAPK/ERK regulated, cytosolic, calcium-dependent, group IVA phospholipase A2 (PLA2G4A) as a novel HCV dependency factor. Inhibition of PLA2G4A activity reduced core protein abundance at lipid droplets, core envelopment and secretion of particles. Moreover, released particles displayed aberrant protein composition and were 100-fold less infectious. Exogenous addition of arachidonic acid, the cleavage product of PLA2G4A-catalyzed lipolysis, but not other related poly-unsaturated fatty acids restored infectivity. Strikingly, production of infectious Dengue virus, a relative of HCV, was also dependent on PLA2G4A. These results highlight previously unrecognized parallels in the assembly pathways of these human pathogens, and define PLA2G4A-dependent lipolysis as crucial prerequisite for production of highly infectious viral progeny.oai:repository.helmholtz-hzi.de:10033/2655132019-08-30T11:24:31Zcom_10033_620589col_10033_620590
Ejaz, Asim
Steinmann, Eike
Bánki, Zoltán
Anggakusuma
Khalid, Sana
Lengauer, Susanne
Wilhelm, Corinne
Zoller, Heinz
Schloegl, Anna
Steinmann, Joerg
Grabski, Elena
Kleines, Michael
Pietschmann, Thomas
Stoiber, Heribert
Institute of Virology, Innsbruck Medical University, Innsbruck, Austria.
2013-01-15T15:33:08Z
2013-01-15T15:33:08Z
2012
Specific acquisition of functional CD59 but not CD46 or CD55 by hepatitis C virus. 2012, 7 (9):e45770 PLoS ONE
1932-6203
23049856
10.1371/journal.pone.0045770
http://hdl.handle.net/10033/265513
PloS one
Viruses of different families encode for regulators of the complement system (RCAs) or acquire such RCAs from the host to get protection against complement-mediated lysis (CML). As hepatitis C virus (HCV) shares no genetic similarity to any known RCA and is detectable at high titers in sera of infected individuals, we investigated whether HCV has adapted host-derived RCAs to resist CML. Here we report that HCV selectively incorporates CD59 while neither CD55, nor CD46 are associated with the virus. The presence of CD59 was shown by capture assays using patient- and cell culture-derived HCV isolates. Association of CD59 with HCV was further confirmed by Western blot analysis using purified viral supernatants from infected Huh 7.5 cells. HCV captured by antibodies specific for CD59 remained infectious for Huh 7.5 cells. In addition, blocking of CD59 in the presence of active complement reduced the titer of HCV most likely due to CML. HCV produced in CD59 knock-down cells were more significantly susceptible to CML compared to wild type virus, but neither replication, assembly nor infectivity of the virus seemed to be impaired in the absence of CD59. In summary our data indicate that HCV incorporates selectively CD59 in its envelope to gain resistance to CML in serum of infected individuals.
en
Archived with thanks to PloS one
Specific acquisition of functional CD59 but not CD46 or CD55 by hepatitis C virus.
Article2018-06-13T09:27:44ZViruses of different families encode for regulators of the complement system (RCAs) or acquire such RCAs from the host to get protection against complement-mediated lysis (CML). As hepatitis C virus (HCV) shares no genetic similarity to any known RCA and is detectable at high titers in sera of infected individuals, we investigated whether HCV has adapted host-derived RCAs to resist CML. Here we report that HCV selectively incorporates CD59 while neither CD55, nor CD46 are associated with the virus. The presence of CD59 was shown by capture assays using patient- and cell culture-derived HCV isolates. Association of CD59 with HCV was further confirmed by Western blot analysis using purified viral supernatants from infected Huh 7.5 cells. HCV captured by antibodies specific for CD59 remained infectious for Huh 7.5 cells. In addition, blocking of CD59 in the presence of active complement reduced the titer of HCV most likely due to CML. HCV produced in CD59 knock-down cells were more significantly susceptible to CML compared to wild type virus, but neither replication, assembly nor infectivity of the virus seemed to be impaired in the absence of CD59. In summary our data indicate that HCV incorporates selectively CD59 in its envelope to gain resistance to CML in serum of infected individuals.oai:repository.helmholtz-hzi.de:10033/2703122019-08-30T11:25:11Zcom_10033_620589col_10033_620590
Fofana, Isabel
Fafi-Kremer, Samira
Carolla, Patric
Fauvelle, Catherine
Zahid, Muhammad Nauman
Turek, Marine
Heydmann, Laura
Cury, Karine
Hayer, Juliette
Combet, Christophe
Cosset, François-Loïc
Pietschmann, Thomas
Hiet, Marie-Sophie
Bartenschlager, Ralf
Habersetzer, François
Doffoël, Michel
Keck, Zhen-Yong
Foung, Steven K H
Zeisel, Mirjam B
Stoll-Keller, Françoise
Baumert, Thomas F
Inserm, U748, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.
2013-02-25T10:38:41Z
2013-02-25T10:38:41Z
2012-07
Mutations that alter use of hepatitis C virus cell entry factors mediate escape from neutralizing antibodies. 2012, 143 (1):223-233.e9 Gastroenterology
1528-0012
22503792
10.1053/j.gastro.2012.04.006
http://hdl.handle.net/10033/270312
Gastroenterology
The development of vaccines and other strategies to prevent hepatitis C virus (HCV) infection is limited by rapid viral evasion. HCV entry is the first step of infection; this process involves several viral and host factors and is targeted by host-neutralizing responses. Although the roles of host factors in HCV entry have been well characterized, their involvement in evasion of immune responses is poorly understood. We used acute infection of liver graft as a model to investigate the molecular mechanisms of viral evasion.
en
Archived with thanks to Gastroenterology
Antibodies, Neutralizing
Cell Line, Tumor
Hepacivirus
Hepatitis C
Humans
Male
Mutation
Transplants
Virus Internalization
Mutations that alter use of hepatitis C virus cell entry factors mediate escape from neutralizing antibodies.
Article2018-06-13T14:13:07ZThe development of vaccines and other strategies to prevent hepatitis C virus (HCV) infection is limited by rapid viral evasion. HCV entry is the first step of infection; this process involves several viral and host factors and is targeted by host-neutralizing responses. Although the roles of host factors in HCV entry have been well characterized, their involvement in evasion of immune responses is poorly understood. We used acute infection of liver graft as a model to investigate the molecular mechanisms of viral evasion.oai:repository.helmholtz-hzi.de:10033/2705122019-08-30T11:33:29Zcom_10033_620589col_10033_620590
Vieyres, Gabrielle
Brohm, Christiane
Friesland, Martina
Gentzsch, Juliane
Wölk, Benno
Roingeard, Philippe
Steinmann, Eike
Pietschmann, Thomas
Institute of Experimental Virology, Twincore, Centre for Experimental and Clinical Infection Research, Hannover, Germany.
2013-02-26T15:43:22Z
2013-02-26T15:43:22Z
2013-02
Subcellular localization and function of an epitope-tagged p7 viroporin in hepatitis C virus-producing cells. 2013, 87 (3):1664-78 J. Virol.
1098-5514
23175364
10.1128/JVI.02782-12
http://hdl.handle.net/10033/270512
Journal of virology
The hepatitis C virus (HCV) viroporin p7 is crucial for production of infectious viral progeny. However, its role in the viral replication cycle remains incompletely understood, in part due to the poor availability of p7-specific antibodies. To circumvent this obstacle, we inserted two consecutive hemagglutinin (HA) epitope tags at its N terminus. HA-tagged p7 reduced peak virus titers ca. 10-fold and decreased kinetics of virus production compared to the wild-type virus. However, HA-tagged p7 rescued virus production of a mutant virus lacking p7, thus providing formal proof that the tag does not disrupt p7 function. In HCV-producing cells, p7 displayed a reticular staining pattern which colocalized with the HCV envelope glycoprotein 2 (E2) but also partially with viral nonstructural proteins 2, 3, and 5A. Using coimmunoprecipitation, we confirmed a specific interaction between p7 and NS2, whereas we did not detect a stable interaction with core, E2, or NS5A. Moreover, we did not observe p7 incorporation into affinity-purified virus particles. Consistently, there was no evidence supporting a role of p7 in viral entry, as an anti-HA antibody was not able to neutralize Jc1 virus produced from an HA-p7-tagged genome. Collectively, these findings highlight a stable interaction between p7 and NS2 which is likely crucial for production of infectious HCV particles. Use of this functional epitope-tagged p7 variant should facilitate the analysis of the final steps of the HCV replication cycle.
en
Archived with thanks to Journal of virology
Subcellular localization and function of an epitope-tagged p7 viroporin in hepatitis C virus-producing cells.
Article2018-06-12T21:29:25ZThe hepatitis C virus (HCV) viroporin p7 is crucial for production of infectious viral progeny. However, its role in the viral replication cycle remains incompletely understood, in part due to the poor availability of p7-specific antibodies. To circumvent this obstacle, we inserted two consecutive hemagglutinin (HA) epitope tags at its N terminus. HA-tagged p7 reduced peak virus titers ca. 10-fold and decreased kinetics of virus production compared to the wild-type virus. However, HA-tagged p7 rescued virus production of a mutant virus lacking p7, thus providing formal proof that the tag does not disrupt p7 function. In HCV-producing cells, p7 displayed a reticular staining pattern which colocalized with the HCV envelope glycoprotein 2 (E2) but also partially with viral nonstructural proteins 2, 3, and 5A. Using coimmunoprecipitation, we confirmed a specific interaction between p7 and NS2, whereas we did not detect a stable interaction with core, E2, or NS5A. Moreover, we did not observe p7 incorporation into affinity-purified virus particles. Consistently, there was no evidence supporting a role of p7 in viral entry, as an anti-HA antibody was not able to neutralize Jc1 virus produced from an HA-p7-tagged genome. Collectively, these findings highlight a stable interaction between p7 and NS2 which is likely crucial for production of infectious HCV particles. Use of this functional epitope-tagged p7 variant should facilitate the analysis of the final steps of the HCV replication cycle.oai:repository.helmholtz-hzi.de:10033/2830332019-08-30T11:35:39Zcom_10033_620589col_10033_620590
Steinmann, J
Buer, J
Pietschmann, T
Steinmann, E
Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. joerg.steinmann@uk-essen.de
2013-04-18T13:14:48Z
2013-04-18T13:14:48Z
2013-03
Anti-infective properties of epigallocatechin-3-gallate (EGCG), a component of green tea. 2013, 168 (5):1059-73 Br. J. Pharmacol.
1476-5381
23072320
10.1111/bph.12009
http://hdl.handle.net/10033/283033
British journal of pharmacology
The consumption of green tea (Camellia sinensis) has been shown to have many physiological and pharmacological health benefits. In the past two decades several studies have reported that epigallocatechin-3-gallate (EGCG), the main constituent of green tea, has anti-infective properties. Antiviral activities of EGCG with different modes of action have been demonstrated on diverse families of viruses, such as Retroviridae, Orthomyxoviridae and Flaviviridae and include important human pathogens like human immunodeficiency virus, influenza A virus and the hepatitis C virus. Furthermore, the molecule interferes with the replication cycle of DNA viruses like hepatitis B virus, herpes simplex virus and adenovirus. Most of these studies demonstrated antiviral properties within physiological concentrations of EGCG in vitro. In contrast, the minimum inhibitory concentrations against bacteria were 10-100-fold higher. Nevertheless, the antibacterial effects of EGCG alone and in combination with different antibiotics have been intensively analysed against a number of bacteria including multidrug-resistant strains such as methicillin-resistant Staphylococcus aureus or Stenotrophomonas maltophilia. Furthermore, the catechin EGCG has antifungal activity against human-pathogenic yeasts like Candida albicans. Although the mechanistic effects of EGCG are not fully understood, there are results indicating that EGCG binds to lipid membranes and affects the folic acid metabolism of bacteria and fungi by inhibiting the cytoplasmic enzyme dihydrofolate reductase. This review summarizes the current knowledge and future perspectives on the antibacterial, antifungal and antiviral effects of the green tea constituent EGCG.
en
Archived with thanks to British journal of pharmacology
Anti-infective properties of epigallocatechin-3-gallate (EGCG), a component of green tea.
Article2014-03-15T00:00:00ZThe consumption of green tea (Camellia sinensis) has been shown to have many physiological and pharmacological health benefits. In the past two decades several studies have reported that epigallocatechin-3-gallate (EGCG), the main constituent of green tea, has anti-infective properties. Antiviral activities of EGCG with different modes of action have been demonstrated on diverse families of viruses, such as Retroviridae, Orthomyxoviridae and Flaviviridae and include important human pathogens like human immunodeficiency virus, influenza A virus and the hepatitis C virus. Furthermore, the molecule interferes with the replication cycle of DNA viruses like hepatitis B virus, herpes simplex virus and adenovirus. Most of these studies demonstrated antiviral properties within physiological concentrations of EGCG in vitro. In contrast, the minimum inhibitory concentrations against bacteria were 10-100-fold higher. Nevertheless, the antibacterial effects of EGCG alone and in combination with different antibiotics have been intensively analysed against a number of bacteria including multidrug-resistant strains such as methicillin-resistant Staphylococcus aureus or Stenotrophomonas maltophilia. Furthermore, the catechin EGCG has antifungal activity against human-pathogenic yeasts like Candida albicans. Although the mechanistic effects of EGCG are not fully understood, there are results indicating that EGCG binds to lipid membranes and affects the folic acid metabolism of bacteria and fungi by inhibiting the cytoplasmic enzyme dihydrofolate reductase. This review summarizes the current knowledge and future perspectives on the antibacterial, antifungal and antiviral effects of the green tea constituent EGCG.oai:repository.helmholtz-hzi.de:10033/2884822019-08-30T11:33:55Zcom_10033_620589col_10033_620590
Gerold, Gisa
Pietschmann, Thomas
Institute of Experimental Virology Twincore-Center for Experimental and Clinical Infectious Disease Research Hannover, Germany.
2013-05-03T12:37:19Z
2013-05-03T12:37:19Z
2013-03
Hepatitis C virus NS5B polymerase primes innate immune signaling. 2013, 57 (3):1275-7 Hepatology
1527-3350
23426794
10.1002/hep.26201
http://hdl.handle.net/10033/288482
Hepatology (Baltimore, Md.)
en
Archived with thanks to Hepatology (Baltimore, Md.)
Hepatitis C virus NS5B polymerase primes innate immune signaling.
Article2014-03-15T00:00:00Zoai:repository.helmholtz-hzi.de:10033/2930052019-08-30T11:24:31Zcom_10033_620589col_10033_620590
Gentzsch, Juliane
Brohm, Christiane
Steinmann, Eike
Friesland, Martina
Menzel, Nicolas
Vieyres, Gabrielle
Perin, Paula Monteiro
Frentzen, Anne
Kaderali, Lars
Pietschmann, Thomas
Institute of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Hannover, Germany.
2013-05-29T10:36:29Z
2013-05-29T10:36:29Z
2013-05
Hepatitis C Virus p7 is Critical for Capsid Assembly and Envelopment. 2013, 9 (5):e1003355 PLoS Pathog.
1553-7374
23658526
10.1371/journal.ppat.1003355
http://hdl.handle.net/10033/293005
PLoS pathogens
Hepatitis C virus (HCV) p7 is a membrane-associated ion channel protein crucial for virus production. To analyze how p7 contributes to this process, we dissected HCV morphogenesis into sub-steps including recruitment of HCV core to lipid droplets (LD), virus capsid assembly, unloading of core protein from LDs and subsequent membrane envelopment of capsids. Interestingly, we observed accumulation of slowly sedimenting capsid-like structures lacking the viral envelope in cells transfected with HCV p7 mutant genomes which possess a defect in virion production. Concomitantly, core protein was enriched at the surface of LDs. This indicates a defect in core/capsid unloading from LDs and subsequent membrane envelopment rather than defective trafficking of core to this cellular organelle. Protease and ribonuclease digestion protection assays, rate zonal centrifugation and native, two dimensional gel electrophoresis revealed increased amounts of high-order, non-enveloped core protein complexes unable to protect viral RNA in cells transfected with p7 mutant genomes. These results suggest accumulation of capsid assembly intermediates that had not yet completely incorporated viral RNA in the absence of functional p7. Thus, functional p7 is necessary for the final steps of capsid assembly as well as for capsid envelopment. These results support a model where capsid assembly is linked with membrane envelopment of nascent RNA-containing core protein multimers, a process coordinated by p7. In summary, we provide novel insights into the sequence of HCV assembly events and essential functions of p7.
en
Archived with thanks to PLoS pathogens
Hepatitis C Virus p7 is Critical for Capsid Assembly and Envelopment.
Article2018-06-12T23:30:07ZHepatitis C virus (HCV) p7 is a membrane-associated ion channel protein crucial for virus production. To analyze how p7 contributes to this process, we dissected HCV morphogenesis into sub-steps including recruitment of HCV core to lipid droplets (LD), virus capsid assembly, unloading of core protein from LDs and subsequent membrane envelopment of capsids. Interestingly, we observed accumulation of slowly sedimenting capsid-like structures lacking the viral envelope in cells transfected with HCV p7 mutant genomes which possess a defect in virion production. Concomitantly, core protein was enriched at the surface of LDs. This indicates a defect in core/capsid unloading from LDs and subsequent membrane envelopment rather than defective trafficking of core to this cellular organelle. Protease and ribonuclease digestion protection assays, rate zonal centrifugation and native, two dimensional gel electrophoresis revealed increased amounts of high-order, non-enveloped core protein complexes unable to protect viral RNA in cells transfected with p7 mutant genomes. These results suggest accumulation of capsid assembly intermediates that had not yet completely incorporated viral RNA in the absence of functional p7. Thus, functional p7 is necessary for the final steps of capsid assembly as well as for capsid envelopment. These results support a model where capsid assembly is linked with membrane envelopment of nascent RNA-containing core protein multimers, a process coordinated by p7. In summary, we provide novel insights into the sequence of HCV assembly events and essential functions of p7.oai:repository.helmholtz-hzi.de:10033/2944392019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Zahid, Muhammad N
Turek, Marine
Xiao, Fei
Thi, Viet Loan Dao
Guérin, Maryse
Fofana, Isabel
Bachellier, Philippe
Thompson, John
Delang, Leen
Neyts, Johan
Bankwitz, Dorothea
Pietschmann, Thomas
Dreux, Marlène
Cosset, François-Loïc
Grunert, Fritz
Baumert, Thomas F
Zeisel, Mirjam B
INSERM, U748, Strasbourg, France.
2013-06-24T09:46:00Z
2013-06-24T09:46:00Z
2013-02
The postbinding activity of scavenger receptor class B type I mediates initiation of hepatitis C virus infection and viral dissemination. 2013, 57 (2):492-504 Hepatology
1527-3350
23081796
10.1002/hep.26097
http://hdl.handle.net/10033/294439
Hepatology (Baltimore, Md.)
Scavenger receptor class B type I (SR-BI) is a high-density lipoprotein (HDL) receptor highly expressed in the liver and modulating HDL metabolism. Hepatitis C virus (HCV) is able to directly interact with SR-BI and requires this receptor to efficiently enter into hepatocytes to establish productive infection. A complex interplay between lipoproteins, SR-BI and HCV envelope glycoproteins has been reported to take place during this process. SR-BI has been demonstrated to act during binding and postbinding steps of HCV entry. Although the SR-BI determinants involved in HCV binding have been partially characterized, the postbinding function of SR-BI remains largely unknown. To uncover the mechanistic role of SR-BI in viral initiation and dissemination, we generated a novel class of anti-SR-BI monoclonal antibodies that interfere with postbinding steps during the HCV entry process without interfering with HCV particle binding to the target cell surface. Using the novel class of antibodies and cell lines expressing murine and human SR-BI, we demonstrate that the postbinding function of SR-BI is of key impact for both initiation of HCV infection and viral dissemination. Interestingly, this postbinding function of SR-BI appears to be unrelated to HDL interaction but to be directly linked to its lipid transfer function.
en
Archived with thanks to Hepatology (Baltimore, Md.)
Animals
Antibodies, Monoclonal
Antigens, CD36
Cell Line
Cholesterol, HDL
Hepacivirus
Hepatitis C
Humans
Lipoproteins, HDL
Mice
Rats
Receptors, Lipoprotein
The postbinding activity of scavenger receptor class B type I mediates initiation of hepatitis C virus infection and viral dissemination.
Article2014-02-15T00:00:00ZScavenger receptor class B type I (SR-BI) is a high-density lipoprotein (HDL) receptor highly expressed in the liver and modulating HDL metabolism. Hepatitis C virus (HCV) is able to directly interact with SR-BI and requires this receptor to efficiently enter into hepatocytes to establish productive infection. A complex interplay between lipoproteins, SR-BI and HCV envelope glycoproteins has been reported to take place during this process. SR-BI has been demonstrated to act during binding and postbinding steps of HCV entry. Although the SR-BI determinants involved in HCV binding have been partially characterized, the postbinding function of SR-BI remains largely unknown. To uncover the mechanistic role of SR-BI in viral initiation and dissemination, we generated a novel class of anti-SR-BI monoclonal antibodies that interfere with postbinding steps during the HCV entry process without interfering with HCV particle binding to the target cell surface. Using the novel class of antibodies and cell lines expressing murine and human SR-BI, we demonstrate that the postbinding function of SR-BI is of key impact for both initiation of HCV infection and viral dissemination. Interestingly, this postbinding function of SR-BI appears to be unrelated to HDL interaction but to be directly linked to its lipid transfer function.oai:repository.helmholtz-hzi.de:10033/2947722019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Schemuth, H
Dittmer, S
Lackner, M
Sedlacek, L
Hamprecht, A
Steinmann, E
Buer, J
Rath, P-M
Steinmann, J
Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Hufelandstrasse 55, Essen, Germany.
2013-06-27T12:29:21Z
2013-06-27T12:29:21Z
2013-05
In vitro activity of colistin as single agent and in combination with antifungals against filamentous fungi occurring in patients with cystic fibrosis. 2013, 56 (3):297-303 Mycoses
1439-0507
23170818
10.1111/myc.12022
http://hdl.handle.net/10033/294772
Mycoses
Because published reports indicate that the antibiotic colistin (COL) has antifungal properties, this study investigated the antifungal in vitro activity of COL as single agent and in combination with the antifungal compounds voriconazole (VRC), caspofungin (CAS) and amphotericin B (AMB) against Scedosporium/Pseudallescheria spp., Exophiala dermatitidis and Geosmithia argillacea. In total, susceptibility was determined for 77 Scedosporium/Pseudallescheria spp., 82 E. dermatitidis and 17 G. argillacea isolates. The minimal inhibitory concentrations (MICs) of COL and the antifungals as single compound and in combination were determined with MIC test strips. Drug interactions were detected by crossing the MIC test strips at a 90º angle. The fractional inhibitory concentration index was used to categorise the drugs' interaction. The MIC50 value of COL was 12 μg ml(-1) for S. prolificans, 16 μg ml(-1) for P. apiosperma, 16 μg ml(-1) for P. boydii, 12 μg ml(-1) for E. dermatiditis and 6 μg ml(-1) for G. argillacea. VRC was the most active drug in combination without any antagonism with the exception of few P. boydii isolates. COL as single agent and in most combinations with antifungals exhibits in vitro antifungal activity against filamentous ascomycetes occurring in cystic fibrosis patients and may offer a novel therapeutic option, especially for multidrug-resistant S. prolificans.
en
Archived with thanks to Mycoses
In vitro activity of colistin as single agent and in combination with antifungals against filamentous fungi occurring in patients with cystic fibrosis.
Article2014-03-15T00:00:00ZBecause published reports indicate that the antibiotic colistin (COL) has antifungal properties, this study investigated the antifungal in vitro activity of COL as single agent and in combination with the antifungal compounds voriconazole (VRC), caspofungin (CAS) and amphotericin B (AMB) against Scedosporium/Pseudallescheria spp., Exophiala dermatitidis and Geosmithia argillacea. In total, susceptibility was determined for 77 Scedosporium/Pseudallescheria spp., 82 E. dermatitidis and 17 G. argillacea isolates. The minimal inhibitory concentrations (MICs) of COL and the antifungals as single compound and in combination were determined with MIC test strips. Drug interactions were detected by crossing the MIC test strips at a 90º angle. The fractional inhibitory concentration index was used to categorise the drugs' interaction. The MIC50 value of COL was 12 μg ml(-1) for S. prolificans, 16 μg ml(-1) for P. apiosperma, 16 μg ml(-1) for P. boydii, 12 μg ml(-1) for E. dermatiditis and 6 μg ml(-1) for G. argillacea. VRC was the most active drug in combination without any antagonism with the exception of few P. boydii isolates. COL as single agent and in most combinations with antifungals exhibits in vitro antifungal activity against filamentous ascomycetes occurring in cystic fibrosis patients and may offer a novel therapeutic option, especially for multidrug-resistant S. prolificans.oai:repository.helmholtz-hzi.de:10033/2959742019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Steinmann, Eike
Pietschmann, Thomas
Helmholtz Centre for Infection Research, Hannover, Germany.
2013-07-15T11:00:42Z
2013-07-15T11:00:42Z
2013
Cell culture systems for hepatitis C virus. 2013, 369:17-48 Curr. Top. Microbiol. Immunol.
0070-217X
23463196
10.1007/978-3-642-27340-7_2
http://hdl.handle.net/10033/295974
Current topics in microbiology and immunology
Due to the obligatory intracellular lifestyle of viruses, cell culture systems for efficient viral propagation are crucial to obtain a detailed understanding of the virus-host cell interaction. For hepatitis C virus (HCV) the development of permissive and authentic culture models continues to be a challenging task. The first efforts to culture HCV had limited success and range back to before the virus was molecularly cloned in 1989. Since then several major breakthroughs have gradually overcome limitations in culturing the virus and sequentially permitted analysis of viral RNA replication, cell entry, and ultimately the complete replication cycle in cultured cells in 2005. Until today, basic and applied HCV research greatly benefit from these tremendous efforts which spurred multiple complementary cell-based model systems for distinct steps of the HCV replication cycle. When used in combination they now permit deep insights into the fascinating biology of HCV and its interplay with the host cell. In fact, drug development has been much facilitated and our understanding of the molecular determinants of HCV replication has grown in parallel to these advances. Building on this groundwork and further refining our cellular models to better mimic the architecture, polarization and differentiation of natural hepatocytes should reveal novel unique aspects of HCV replication. Ultimately, models to culture primary HCV isolates across all genotypes may teach us important new lessons about viral functional adaptations that have evolved in exchange with its human host and that may explain the variable natural course of hepatitis C.
en
Archived with thanks to Current topics in microbiology and immunology
Cell culture systems for hepatitis C virus.
Book chapter2014-03-15T00:00:00ZDue to the obligatory intracellular lifestyle of viruses, cell culture systems for efficient viral propagation are crucial to obtain a detailed understanding of the virus-host cell interaction. For hepatitis C virus (HCV) the development of permissive and authentic culture models continues to be a challenging task. The first efforts to culture HCV had limited success and range back to before the virus was molecularly cloned in 1989. Since then several major breakthroughs have gradually overcome limitations in culturing the virus and sequentially permitted analysis of viral RNA replication, cell entry, and ultimately the complete replication cycle in cultured cells in 2005. Until today, basic and applied HCV research greatly benefit from these tremendous efforts which spurred multiple complementary cell-based model systems for distinct steps of the HCV replication cycle. When used in combination they now permit deep insights into the fascinating biology of HCV and its interplay with the host cell. In fact, drug development has been much facilitated and our understanding of the molecular determinants of HCV replication has grown in parallel to these advances. Building on this groundwork and further refining our cellular models to better mimic the architecture, polarization and differentiation of natural hepatocytes should reveal novel unique aspects of HCV replication. Ultimately, models to culture primary HCV isolates across all genotypes may teach us important new lessons about viral functional adaptations that have evolved in exchange with its human host and that may explain the variable natural course of hepatitis C.oai:repository.helmholtz-hzi.de:10033/2968372019-08-30T11:25:11Zcom_10033_620589col_10033_620590
Westhaus, Sandra
Bankwitz, Dorothea
Ernst, Stefanie
Rohrmann, Katrin
Wappler, Ilka
Agné, Clemens
Luchtefeld, Maren
Schieffer, Bernhard
Sarrazin, Christoph
Manns, Michael P
Pietschmann, Thomas
Ciesek, Sandra
von Hahn, Thomas
Institute for Molecular Biology, Medizinische Hochschule Hannover, Hannover, Germany.
2013-07-23T09:33:06Z
2013-07-23T09:33:06Z
2013-05
Characterization of the inhibition of hepatitis C virus entry by in vitro-generated and patient-derived oxidized low-density lipoprotein. 2013, 57 (5):1716-24 Hepatology
1527-3350
23212706
10.1002/hep.26190
http://hdl.handle.net/10033/296837
Hepatology (Baltimore, Md.)
Oxidized low-density lipoprotein (oxLDL) has been reported as an inhibitor of hepatitis C virus (HCV) cell entry, making it the only known component of human lipid metabolism with an antiviral effect on HCV. However, several questions remain open, including its effect on full-length cell-culture-grown HCV (HCVcc) of different genotypes or on other steps of the viral replication cycle, its mechanism of action, and whether endogenous oxLDL shares the anti-HCV properties of in vitro-generated oxLDL. We combined molecular virology tools with oxLDL serum measurements in different patient cohorts to address these questions. We found that oxLDL inhibits HCVcc at least as potently as HCV pseudoparticles. There was moderate variation between genotypes, with genotype 4 appearing the most oxLDL sensitive. Intracellular RNA replication and assembly and release of new particles were unaffected. HCV particles entering target cells lost oxLDL sensitivity with time kinetics parallel to anti-SR-BI (scavenger receptor class B type I), but significantly earlier than anti-CD81, suggesting that oxLDL acts by perturbing interaction between HCV and SR-BI. Finally, in chronically HCV-infected individuals, endogenous serum oxLDL levels did not correlate with viral load, but in HCV-negative sera, high endogenous oxLDL had a negative effect on HCV infectivity in vitro. Conclusion: oxLDL is a potent pangenotype HCV entry inhibitor that maintains its activity in the context of human serum and targets an early step of HCV entry.
en
Archived with thanks to Hepatology (Baltimore, Md.)
Antigens, CD36
Carcinoma, Hepatocellular
Cell Line, Tumor
Cells, Cultured
DNA, Viral
Genotype
Hepacivirus
Hepatitis C, Chronic
Humans
Lipoproteins, LDL
Liver Neoplasms
Viral Load
Virion
Virus Replication
Characterization of the inhibition of hepatitis C virus entry by in vitro-generated and patient-derived oxidized low-density lipoprotein.
Article2014-05-15T00:00:00ZOxidized low-density lipoprotein (oxLDL) has been reported as an inhibitor of hepatitis C virus (HCV) cell entry, making it the only known component of human lipid metabolism with an antiviral effect on HCV. However, several questions remain open, including its effect on full-length cell-culture-grown HCV (HCVcc) of different genotypes or on other steps of the viral replication cycle, its mechanism of action, and whether endogenous oxLDL shares the anti-HCV properties of in vitro-generated oxLDL. We combined molecular virology tools with oxLDL serum measurements in different patient cohorts to address these questions. We found that oxLDL inhibits HCVcc at least as potently as HCV pseudoparticles. There was moderate variation between genotypes, with genotype 4 appearing the most oxLDL sensitive. Intracellular RNA replication and assembly and release of new particles were unaffected. HCV particles entering target cells lost oxLDL sensitivity with time kinetics parallel to anti-SR-BI (scavenger receptor class B type I), but significantly earlier than anti-CD81, suggesting that oxLDL acts by perturbing interaction between HCV and SR-BI. Finally, in chronically HCV-infected individuals, endogenous serum oxLDL levels did not correlate with viral load, but in HCV-negative sera, high endogenous oxLDL had a negative effect on HCV infectivity in vitro. Conclusion: oxLDL is a potent pangenotype HCV entry inhibitor that maintains its activity in the context of human serum and targets an early step of HCV entry.oai:repository.helmholtz-hzi.de:10033/2980672019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Doerrbecker, J
Meuleman, P
Kang, J
Riebesehl, N
Wilhelm, C
Friesland, M
Pfaender, S
Steinmann, J
Pietschmann, T
Steinmann, E
Division of Experimental Virology, Twincore Center for Experimental and Clinical Infection Research, Feodor-Lynen-Straße 7-9, 30625 Hannover, Germany.
2013-08-13T09:48:31Z
2013-08-13T09:48:31Z
2013-07
Thermostability of seven hepatitis C virus genotypes in vitro and in vivo. 2013, 20 (7):478-85 J. Viral Hepat.
1365-2893
23730841
10.1111/jvh.12055
http://hdl.handle.net/10033/298067
Journal of viral hepatitis
Hepatitis C virus (HCV) is transmitted primarily through percutaneous exposure to contaminated blood especially in healthcare settings and among people who inject drugs. The environmental stability of HCV has been extrapolated from studies with the bovine viral diarrhoea virus or was so far only addressed with HCV genotype 2a viruses. The aim of this study was to compare the environmental and thermostability of all so far known seven HCV genotypes in vitro and in vivo. Incubation experiments at room temperature revealed that all HCV genotypes showed similar environmental stabilities in suspension with viral infectivity detectable for up to 28 days. The risk of HCV infection may not accurately be reflected by determination of HCV RNA levels. However, viral stability and transmission risks assessed from in vitro experiments correlated with viral infectivity in transgenic mice containing human liver xenografts. A reduced viral stability for up to 2 days was observed at 37 °C with comparable decays for all HCV genotypes confirmed by thermodynamic analysis. These results demonstrate that different HCV genotypes possess comparable stability in the environment and that noninfectious particles after incubation in vitro do not cause infection in an HCV in vivo model. These findings are important for estimation of HCV cross-transmission in the environment and indicate that different HCV genotypes do not display an altered stability or resistance at certain temperatures.
en
Archived with thanks to Journal of viral hepatitis
Thermostability of seven hepatitis C virus genotypes in vitro and in vivo.
Article2014-07-15T00:00:00ZHepatitis C virus (HCV) is transmitted primarily through percutaneous exposure to contaminated blood especially in healthcare settings and among people who inject drugs. The environmental stability of HCV has been extrapolated from studies with the bovine viral diarrhoea virus or was so far only addressed with HCV genotype 2a viruses. The aim of this study was to compare the environmental and thermostability of all so far known seven HCV genotypes in vitro and in vivo. Incubation experiments at room temperature revealed that all HCV genotypes showed similar environmental stabilities in suspension with viral infectivity detectable for up to 28 days. The risk of HCV infection may not accurately be reflected by determination of HCV RNA levels. However, viral stability and transmission risks assessed from in vitro experiments correlated with viral infectivity in transgenic mice containing human liver xenografts. A reduced viral stability for up to 2 days was observed at 37 °C with comparable decays for all HCV genotypes confirmed by thermodynamic analysis. These results demonstrate that different HCV genotypes possess comparable stability in the environment and that noninfectious particles after incubation in vitro do not cause infection in an HCV in vivo model. These findings are important for estimation of HCV cross-transmission in the environment and indicate that different HCV genotypes do not display an altered stability or resistance at certain temperatures.oai:repository.helmholtz-hzi.de:10033/2990862019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Vieyres, Gabrielle
Pietschmann, Thomas
Institute of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research; A Joint Venture Between The Medical School Hannover and The Helmholtz Centre for Infection Research, Feodor-Lynen-Straße 7-9, 30625 Hannover, Germany.
2013-08-19T12:21:26Z
2013-08-19T12:21:26Z
2013-02
Entry and replication of recombinant hepatitis C viruses in cell culture. 2013, 59 (2):233-48 Methods
1095-9130
23009812
10.1016/j.ymeth.2012.09.005
http://hdl.handle.net/10033/299086
Methods (San Diego, Calif.)
Hepatitis C virus (HCV) is a positive-strand enveloped RNA virus and belongs to the Flaviviridae family. The heavy health burden associated with the virus infection in humans and the intriguing peculiarities of the interaction between the HCV replication cycle and the hepatocyte host cell have stimulated a flourishing research field. The present review aims at recapitulating the different viral and cellular systems modelling HCV entry and replication, and in particular at gathering the tools available to dissect the HCV entry pathway.
en
Archived with thanks to Methods (San Diego, Calif.)
Entry and replication of recombinant hepatitis C viruses in cell culture.
Article2018-06-12T23:06:36ZHepatitis C virus (HCV) is a positive-strand enveloped RNA virus and belongs to the Flaviviridae family. The heavy health burden associated with the virus infection in humans and the intriguing peculiarities of the interaction between the HCV replication cycle and the hepatocyte host cell have stimulated a flourishing research field. The present review aims at recapitulating the different viral and cellular systems modelling HCV entry and replication, and in particular at gathering the tools available to dissect the HCV entry pathway.oai:repository.helmholtz-hzi.de:10033/3054922019-08-30T11:27:16Zcom_10033_620589col_10033_620590
Saeed, A
Baloch, K
Brown, R J P
Wallis, R
Chen, L
Dexter, L
McClure, C P
Shakesheff, K
Thomson, B J
School of Molecular Medical Sciences, University of Nottingham, Leicester, UK; School of Pharmacy, University of Nottingham, Leicester, UK.
2013-11-18T14:13:32Z
2013-11-18T14:13:32Z
2013-11
Mannan binding lectin-associated serine protease 1 is induced by hepatitis C virus infection and activates human hepatic stellate cells. 2013, 174 (2):265-73 Clin. Exp. Immunol.
1365-2249
23841802
10.1111/cei.12174
http://hdl.handle.net/10033/305492
Clinical and experimental immunology
Mannan binding lectin (MBL)-associated serine protease type 1 (MASP-1) has a central role in the lectin pathway of complement activation and is required for the formation of C3 convertase. The activity of MASP-1 in the peripheral blood has been identified previously as a highly significant predictor of the severity of liver fibrosis in hepatitis C virus (HCV) infection, but not in liver disease of other aetiologies. In this study we tested the hypotheses that expression of MASP-1 may promote disease progression in HCV disease by direct activation of hepatic stellate cells (HSCs) and may additionally be up-regulated by HCV. In order to do so, we utilized a model for the maintenance of primary human HSC in the quiescent state by culture on basement membrane substrate prior to stimulation. In comparison to controls, recombinant MASP-1 stimulated quiescent human HSCs to differentiate to the activated state as assessed by both morphology and up-regulation of HSC activation markers α-smooth muscle actin and tissue inhibitor of metalloproteinase 1. Further, the expression of MASP-1 was up-regulated significantly by HCV infection in hepatocyte cell lines. These observations suggest a new role for MASP-1 and provide a possible mechanistic link between high levels of MASP-1 and the severity of disease in HCV infection. Taken together with previous clinical observations, our new findings suggest that the balance of MASP-1 activity may be proinflammatory and act to accelerate fibrosis progression in HCV liver disease.
en
Archived with thanks to Clinical and experimental immunology
Mannan binding lectin-associated serine protease 1 is induced by hepatitis C virus infection and activates human hepatic stellate cells.
Article2014-11-15T00:00:00ZMannan binding lectin (MBL)-associated serine protease type 1 (MASP-1) has a central role in the lectin pathway of complement activation and is required for the formation of C3 convertase. The activity of MASP-1 in the peripheral blood has been identified previously as a highly significant predictor of the severity of liver fibrosis in hepatitis C virus (HCV) infection, but not in liver disease of other aetiologies. In this study we tested the hypotheses that expression of MASP-1 may promote disease progression in HCV disease by direct activation of hepatic stellate cells (HSCs) and may additionally be up-regulated by HCV. In order to do so, we utilized a model for the maintenance of primary human HSC in the quiescent state by culture on basement membrane substrate prior to stimulation. In comparison to controls, recombinant MASP-1 stimulated quiescent human HSCs to differentiate to the activated state as assessed by both morphology and up-regulation of HSC activation markers α-smooth muscle actin and tissue inhibitor of metalloproteinase 1. Further, the expression of MASP-1 was up-regulated significantly by HCV infection in hepatocyte cell lines. These observations suggest a new role for MASP-1 and provide a possible mechanistic link between high levels of MASP-1 and the severity of disease in HCV infection. Taken together with previous clinical observations, our new findings suggest that the balance of MASP-1 activity may be proinflammatory and act to accelerate fibrosis progression in HCV liver disease.oai:repository.helmholtz-hzi.de:10033/3054932019-08-30T11:34:19Zcom_10033_620589col_10033_620590
Behrendt, Patrick
Doerrbecker, Juliane
Riebesehl, Nina
Wilhelm, Corinne
Ciesek, Sandra
Erichsen, Thomas J
Steinmann, Joerg
Ott, Michael
Manns, Michael P
Pietschmann, Thomas
Steinmann, Eike
Division of Experimental Virology, Twincore, Center for Experimental and Clinical Infection Research, Hannover, Germany; Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
2013-11-18T14:51:09Z
2013-11-18T14:51:09Z
2013-10
Stability and transmission of hepatitis C virus in different anesthetic agents. 2013, 41 (10):942-3 Am J Infect Control
1527-3296
23523523
10.1016/j.ajic.2013.01.016
http://hdl.handle.net/10033/305493
American journal of infection control
en
Archived with thanks to American journal of infection control
Stability and transmission of hepatitis C virus in different anesthetic agents.
article2018-06-13T01:07:43Zoai:repository.helmholtz-hzi.de:10033/3057032019-08-30T11:25:11Zcom_10033_620589col_10033_620590
Tarr, Alexander W
Lafaye, Pierre
Meredith, Luke
Damier-Piolle, Laurence
Urbanowicz, Richard A
Meola, Annalisa
Jestin, Jean-Luc
Brown, Richard J P
McKeating, Jane A
Rey, Felix A
Ball, Jonathan K
Krey, Thomas
School of Molecular Medical Sciences, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom.
2013-11-22T15:45:21Z
2013-11-22T15:45:21Z
2013-09
An alpaca nanobody inhibits hepatitis C virus entry and cell-to-cell transmission. 2013, 58 (3):932-9 Hepatology
1527-3350
23553604
10.1002/hep.26430
http://hdl.handle.net/10033/305703
Hepatology (Baltimore, Md.)
Severe liver disease caused by chronic hepatitis C virus is the major indication for liver transplantation. Despite recent advances in antiviral therapy, drug toxicity and unwanted side effects render effective treatment in liver-transplanted patients a challenging task. Virus-specific therapeutic antibodies are generally safe and well-tolerated, but their potential in preventing and treating hepatitis C virus (HCV) infection has not yet been realized due to a variety of issues, not least high production costs and virus variability. Heavy-chain antibodies or nanobodies, produced by camelids, represent an exciting antiviral approach; they can target novel highly conserved epitopes that are inaccessible to normal antibodies, and they are also easy to manipulate and produce. We isolated four distinct nanobodies from a phage-display library generated from an alpaca immunized with HCV E2 glycoprotein. One of them, nanobody D03, recognized a novel epitope overlapping with the epitopes of several broadly neutralizing human monoclonal antibodies. Its crystal structure revealed a long complementarity determining region (CD3) folding over part of the framework that, in conventional antibodies, forms the interface between heavy and light chain. D03 neutralized a panel of retroviral particles pseudotyped with HCV glycoproteins from six genotypes and authentic cell culture-derived particles by interfering with the E2-CD81 interaction. In contrast to some of the most broadly neutralizing human anti-E2 monoclonal antibodies, D03 efficiently inhibited HCV cell-to-cell transmission. Conclusion: This is the first description of a potent and broadly neutralizing HCV-specific nanobody representing a significant advance that will lead to future development of novel entry inhibitors for the treatment and prevention of HCV infection and help our understanding of HCV cell-to-cell transmission.
en
Archived with thanks to Hepatology (Baltimore, Md.)
An alpaca nanobody inhibits hepatitis C virus entry and cell-to-cell transmission.
Article2014-10-15T00:00:00ZSevere liver disease caused by chronic hepatitis C virus is the major indication for liver transplantation. Despite recent advances in antiviral therapy, drug toxicity and unwanted side effects render effective treatment in liver-transplanted patients a challenging task. Virus-specific therapeutic antibodies are generally safe and well-tolerated, but their potential in preventing and treating hepatitis C virus (HCV) infection has not yet been realized due to a variety of issues, not least high production costs and virus variability. Heavy-chain antibodies or nanobodies, produced by camelids, represent an exciting antiviral approach; they can target novel highly conserved epitopes that are inaccessible to normal antibodies, and they are also easy to manipulate and produce. We isolated four distinct nanobodies from a phage-display library generated from an alpaca immunized with HCV E2 glycoprotein. One of them, nanobody D03, recognized a novel epitope overlapping with the epitopes of several broadly neutralizing human monoclonal antibodies. Its crystal structure revealed a long complementarity determining region (CD3) folding over part of the framework that, in conventional antibodies, forms the interface between heavy and light chain. D03 neutralized a panel of retroviral particles pseudotyped with HCV glycoproteins from six genotypes and authentic cell culture-derived particles by interfering with the E2-CD81 interaction. In contrast to some of the most broadly neutralizing human anti-E2 monoclonal antibodies, D03 efficiently inhibited HCV cell-to-cell transmission. Conclusion: This is the first description of a potent and broadly neutralizing HCV-specific nanobody representing a significant advance that will lead to future development of novel entry inhibitors for the treatment and prevention of HCV infection and help our understanding of HCV cell-to-cell transmission.oai:repository.helmholtz-hzi.de:10033/3234302019-08-30T11:24:31Zcom_10033_620589col_10033_620590
Lange, Milena
Fiedler, Melanie
Bankwitz, Dorothea
Osburn, William
Viazov, Sergei
Brovko, Olena
Zekri, Abdel-Rahman
Khudyakov, Yury
Nassal, Michael
Pumpens, Paul
Pietschmann, Thomas
Timm, Jörg
Roggendorf, Michael
Walker, Andreas
2014-07-18T13:24:25Z
2014-07-18T13:24:25Z
2014
Hepatitis C virus hypervariable region 1 variants presented on hepatitis B virus capsid-like particles induce cross-neutralizing antibodies. 2014, 9 (7):e102235 PLoS ONE
1932-6203
25014219
10.1371/journal.pone.0102235
http://hdl.handle.net/10033/323430
PloS one
Hepatitis C virus (HCV) infection is still a serious global health burden. Despite improved therapeutic options, a preventative vaccine would be desirable especially in undeveloped countries. Traditionally, highly conserved epitopes are targets for antibody-based prophylactic vaccines. In HCV-infected patients, however, neutralizing antibodies are primarily directed against hypervariable region I (HVRI) in the envelope protein E2. HVRI is the most variable region of HCV, and this heterogeneity contributes to viral persistence and has thus far prevented the development of an effective HVRI-based vaccine. The primary goal of an antibody-based HCV vaccine should therefore be the induction of cross-reactive HVRI antibodies. In this study we approached this problem by presenting selected cross-reactive HVRI variants in a highly symmetric repeated array on capsid-like particles (CLPs). SplitCore CLPs, a novel particulate antigen presentation system derived from the HBV core protein, were used to deliberately manipulate the orientation of HVRI and therefore enable the presentation of conserved parts of HVRI. These HVRI-CLPs induced high titers of cross-reactive antibodies, including neutralizing antibodies. The combination of only four HVRI CLPs was sufficient to induce antibodies cross-reactive with 81 of 326 (24.8%) naturally occurring HVRI peptides. Most importantly, HVRI CLPs with AS03 as an adjuvant induced antibodies with a 10-fold increase in neutralizing capability. These antibodies were able to neutralize infectious HCVcc isolates and 4 of 19 (21%) patient-derived HCVpp isolates. Taken together, these results demonstrate that the induction of at least partially cross-neutralizing antibodies is possible. This approach might be useful for the development of a prophylactic HCV vaccine and should also be adaptable to other highly variable viruses.
en
Archived with thanks to PloS one
Hepatitis C virus hypervariable region 1 variants presented on hepatitis B virus capsid-like particles induce cross-neutralizing antibodies.
Article2018-06-13T05:28:47ZHepatitis C virus (HCV) infection is still a serious global health burden. Despite improved therapeutic options, a preventative vaccine would be desirable especially in undeveloped countries. Traditionally, highly conserved epitopes are targets for antibody-based prophylactic vaccines. In HCV-infected patients, however, neutralizing antibodies are primarily directed against hypervariable region I (HVRI) in the envelope protein E2. HVRI is the most variable region of HCV, and this heterogeneity contributes to viral persistence and has thus far prevented the development of an effective HVRI-based vaccine. The primary goal of an antibody-based HCV vaccine should therefore be the induction of cross-reactive HVRI antibodies. In this study we approached this problem by presenting selected cross-reactive HVRI variants in a highly symmetric repeated array on capsid-like particles (CLPs). SplitCore CLPs, a novel particulate antigen presentation system derived from the HBV core protein, were used to deliberately manipulate the orientation of HVRI and therefore enable the presentation of conserved parts of HVRI. These HVRI-CLPs induced high titers of cross-reactive antibodies, including neutralizing antibodies. The combination of only four HVRI CLPs was sufficient to induce antibodies cross-reactive with 81 of 326 (24.8%) naturally occurring HVRI peptides. Most importantly, HVRI CLPs with AS03 as an adjuvant induced antibodies with a 10-fold increase in neutralizing capability. These antibodies were able to neutralize infectious HCVcc isolates and 4 of 19 (21%) patient-derived HCVpp isolates. Taken together, these results demonstrate that the induction of at least partially cross-neutralizing antibodies is possible. This approach might be useful for the development of a prophylactic HCV vaccine and should also be adaptable to other highly variable viruses.oai:repository.helmholtz-hzi.de:10033/3254832019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Perales, Celia
Beach, Nathan M
Sheldon, Julie
Domingo, Esteban
2014-08-27T12:51:47Z
2014-08-27T12:51:47Z
2014-06-23
Molecular basis of interferon resistance in hepatitis C virus. 2014, 8C:38-44 Curr Opin Virol
1879-6265
24968186
10.1016/j.coviro.2014.05.003
http://hdl.handle.net/10033/325483
Current opinion in virology
Resistance to interferon (IFN) in hepatitis C virus (HCV) differs from resistance to standard, directly-acting antiviral (DAA) agents in that the virus confronts a multicomponent antiviral state evoked by IFN. This renders unlikely the repeated selection of the same specific mutations that confer an IFN-resistance phenotype. Comparison of amino acid sequences of viral proteins in HCV that replicates in the presence of IFN in vivo or in cell culture (with entire virus or subgenomic replicons) reveals very few common candidate IFN resistance substitutions. Multiple host and viral factors contribute to divergent responses to IFN. The environmental heterogeneity in which exogenous IFN is expected to exert its selective effect may increase as a result of incorporation of new DAAs in therapy.
ENG
Archived with thanks to Current opinion in virology
Molecular basis of interferon resistance in hepatitis C virus.
Article2018-06-12T23:15:39ZResistance to interferon (IFN) in hepatitis C virus (HCV) differs from resistance to standard, directly-acting antiviral (DAA) agents in that the virus confronts a multicomponent antiviral state evoked by IFN. This renders unlikely the repeated selection of the same specific mutations that confer an IFN-resistance phenotype. Comparison of amino acid sequences of viral proteins in HCV that replicates in the presence of IFN in vivo or in cell culture (with entire virus or subgenomic replicons) reveals very few common candidate IFN resistance substitutions. Multiple host and viral factors contribute to divergent responses to IFN. The environmental heterogeneity in which exogenous IFN is expected to exert its selective effect may increase as a result of incorporation of new DAAs in therapy.oai:repository.helmholtz-hzi.de:10033/3258982019-08-30T11:24:24Zcom_10033_620589col_10033_620590
Gerold, Gisa
Pietschmann, Thomas
TWINCORE – Institute of Experimental Virology, Centre for Experimental and Clinical Infection Research, Hannover , Germany
2014-09-05T12:45:52Z
2014-09-05T12:45:52Z
2014-09-05
The HCV Life Cycle: In vitro Tissue Culture Systems and Therapeutic Targets 2014, 32 (5):525 Digestive Diseases
1421-9875
0257-2753
10.1159/000360830
http://hdl.handle.net/10033/325898
Digestive Diseases
Karger
http://www.karger.com?doi=10.1159/000360830
Archived with thanks to Digestive Diseases
The HCV Life Cycle: In vitro Tissue Culture Systems and Therapeutic Targets
Article2018-06-12T22:30:27Zoai:repository.helmholtz-hzi.de:10033/3261732019-08-30T11:34:20Zcom_10033_620589col_10033_620590
Irving, William L.
Rupp, Daniel
McClure, C. Patrick
Than, Lwin Mar
Titman, Andrew
Ball, Jonathan K.
Steinmann, Eike
Bartenschlager, Ralf
Pietschmann, Thomas
Brown, Richard J.P.
2014-09-16T12:19:23Z
2014-09-16T12:19:23Z
2014-09-16
Development of a high-throughput pyrosequencing assay for monitoring temporal evolution and resistance associated variant emergence in the Hepatitis C virus protease coding-region 2014, 110:52 Antiviral Research
01663542
10.1016/j.antiviral.2014.07.009
http://hdl.handle.net/10033/326173
Antiviral Research
http://linkinghub.elsevier.com/retrieve/pii/S0166354214002083
Archived with thanks to Antiviral Research
Development of a high-throughput pyrosequencing assay for monitoring temporal evolution and resistance associated variant emergence in the Hepatitis C virus protease coding-region
Article2018-06-13T02:41:24Zoai:repository.helmholtz-hzi.de:10033/3328652019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Vidigal, P G
Dittmer, S
Steinmann, E
Buer, J
Rath, P-M
Steinmann, J
2014-10-17T13:07:04Z
2014-10-17T13:07:04Z
2014-07
Adaptation of Stenotrophomonas maltophilia in cystic fibrosis: molecular diversity, mutation frequency and antibiotic resistance. 2014, 304 (5-6):613-9 Int. J. Med. Microbiol.
1618-0607
24836944
10.1016/j.ijmm.2014.04.002
http://hdl.handle.net/10033/332865
International journal of medical microbiology : IJMM
Due to the continuous exposure to a challenging environment and repeated antibiotic treatment courses, bacterial populations in cystic fibrosis (CF) patients experience selective pressure causing the emergence of mutator phenotypes. In this study we investigated the genotypic diversity, mutation frequency and antibiotic resistance of S. maltophilia isolates chronically colonizing CF patients. S. maltophilia was isolated from a total of 90 sputum samples, collected sequentially from 19 CF patients admitted between January 2008 and March 2012 at the University Hospital Essen, Germany. DNA fingerprinting by repetitive-sequence-based PCR revealed that 68.4% (n=13) of CF patients harbored different S. maltophilia genotypes during the 4-year study course. Out of 90 S. maltophilia isolates obtained from chronically colonized CF patients, 17.8% (n=16) were hypomutators, 27.7% (n=25), normomutators, 23.3% (n=21), weak hypermutators and 31.2% (n=28) strong hypermutators. We also found that mutation rates of the most clonally related genotypes varied over time with the tendency to become less mutable. Mutator isolates were found to have no significant increase in resistance against eight different antibiotics versus nonmutators. Sequencing of the mismatch repair genes mutL, mutS and uvrD revealed alterations that resulted in amino acid changes in their corresponding proteins. Here, we could demonstrate that several different S. maltophilia genotypes are present in CF patients and as a sign of adaption their mutation status switches over time to a less mutator phenotype without increasing resistance. These results suggest that S. maltophilia attempts to sustain its biological fitness as mechanism for long-term persistence in the CF lung.
en
Archived with thanks to International journal of medical microbiology : IJMM
Adaptation of Stenotrophomonas maltophilia in cystic fibrosis: molecular diversity, mutation frequency and antibiotic resistance.
Article2018-06-13T00:18:54ZDue to the continuous exposure to a challenging environment and repeated antibiotic treatment courses, bacterial populations in cystic fibrosis (CF) patients experience selective pressure causing the emergence of mutator phenotypes. In this study we investigated the genotypic diversity, mutation frequency and antibiotic resistance of S. maltophilia isolates chronically colonizing CF patients. S. maltophilia was isolated from a total of 90 sputum samples, collected sequentially from 19 CF patients admitted between January 2008 and March 2012 at the University Hospital Essen, Germany. DNA fingerprinting by repetitive-sequence-based PCR revealed that 68.4% (n=13) of CF patients harbored different S. maltophilia genotypes during the 4-year study course. Out of 90 S. maltophilia isolates obtained from chronically colonized CF patients, 17.8% (n=16) were hypomutators, 27.7% (n=25), normomutators, 23.3% (n=21), weak hypermutators and 31.2% (n=28) strong hypermutators. We also found that mutation rates of the most clonally related genotypes varied over time with the tendency to become less mutable. Mutator isolates were found to have no significant increase in resistance against eight different antibiotics versus nonmutators. Sequencing of the mismatch repair genes mutL, mutS and uvrD revealed alterations that resulted in amino acid changes in their corresponding proteins. Here, we could demonstrate that several different S. maltophilia genotypes are present in CF patients and as a sign of adaption their mutation status switches over time to a less mutator phenotype without increasing resistance. These results suggest that S. maltophilia attempts to sustain its biological fitness as mechanism for long-term persistence in the CF lung.oai:repository.helmholtz-hzi.de:10033/3330302019-08-30T11:33:30Zcom_10033_620589col_10033_620590
Rath, P-M
Schoch, B
Adamzik, M
Steinmann, E
Buer, J
Steinmann, J
2014-10-21T14:16:32Z
2014-10-21T14:16:32Z
2014-08
Value of multiplex PCR using cerebrospinal fluid for the diagnosis of ventriculostomy-related meningitis in neurosurgery patients. 2014, 42 (4):621-7 Infection
1439-0973
24470322
10.1007/s15010-014-0590-8
http://hdl.handle.net/10033/333030
Infection
This prospective observational cohort study assessed the use of a multiplex real-time polymerase chain reaction (PCR) assay alone and in conjunction with biomarkers for the diagnosis of ventriculostomy-related meningitis in neurosurgery intensive care unit (ICU) patients with external ventricular drainage (EVD).
en
Archived with thanks to Infection
Value of multiplex PCR using cerebrospinal fluid for the diagnosis of ventriculostomy-related meningitis in neurosurgery patients.
Article2015-08-15T00:00:00ZThis prospective observational cohort study assessed the use of a multiplex real-time polymerase chain reaction (PCR) assay alone and in conjunction with biomarkers for the diagnosis of ventriculostomy-related meningitis in neurosurgery intensive care unit (ICU) patients with external ventricular drainage (EVD).oai:repository.helmholtz-hzi.de:10033/3370492019-08-30T11:27:16Zcom_10033_620589col_10033_620590
Pischke, Sven
Behrendt, Patrick
Bock, Claus-Thomas
Jilg, Wolfgang
Manns, Michael P
Wedemeyer, Heiner
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hanover, German Centre for Infection Research, Hanover, TWINCORE Institute for Experimental Infection Research, Hanover, Health Care Center at the University Medical Center Hamburg-Eppendorf, Robert Koch Institute Berlin, Consiliary Laboratory for Hepatitis A and Hepatitis E, Institute for Medical Microbiology and Hygiene, University Hospital Regensburg, Regensburg.
2014-12-11T09:23:59Z
2014-12-11T09:23:59Z
2014-09-01
Hepatitis E in Germany--an under-reported infectious disease. 2014, 111 (35-36):577-83 Dtsch Arztebl Int
1866-0452
25249359
10.3238/arztebl.2014.0577
http://hdl.handle.net/10033/337049
Deutsches Ärzteblatt international
At least 17% of the population in Germany has been infected with the hepatitis E virus (HEV); thus, HEV infections are more frequent than was previously assumed. However, fewer than 500 HEV infections were reported to the Robert Koch Institute in 2013.
en
Hepatitis E in Germany--an under-reported infectious disease.
Article2018-06-12T17:42:46ZAt least 17% of the population in Germany has been infected with the hepatitis E virus (HEV); thus, HEV infections are more frequent than was previously assumed. However, fewer than 500 HEV infections were reported to the Robert Koch Institute in 2013.oai:repository.helmholtz-hzi.de:10033/3454092019-08-30T11:27:16Zcom_10033_620589col_10033_620590
Ciesek, Sandra
von Hahn, Thomas
Colpitts, Che C
Schang, Luis M
Friesland, Martina
Steinmann, Jörg
Manns, Michael P
Ott, Michael
Wedemeyer, Heiner
Meuleman, Philip
Pietschmann, Thomas
Steinmann, Eike
2015-02-26T12:17:23Z
2015-02-26T12:17:23Z
2011-12
The green tea polyphenol, epigallocatechin-3-gallate, inhibits hepatitis C virus entry. 2011, 54 (6):1947-55 Hepatology
1527-3350
21837753
10.1002/hep.24610
http://hdl.handle.net/10033/345409
Hepatology (Baltimore, Md.)
Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. Current antiviral therapy fails to clear infection in a substantial proportion of cases. Drug development is focused on nonstructural proteins required for RNA replication. Individuals undergoing orthotopic liver transplantation face rapid, universal reinfection of the graft. Therefore, antiviral strategies targeting the early stages of infection are urgently needed for the prevention of HCV infection. In this study, we identified the polyphenol, epigallocatechin-3-gallate (EGCG), as an inhibitor of HCV entry. Green tea catechins, such as EGCG and its derivatives, epigallocatechin (EGC), epicatechin gallate (ECG), and epicatechin (EC), have been previously found to exert antiviral and antioncogenic properties. EGCG had no effect on HCV RNA replication, assembly, or release of progeny virions. However, it potently inhibited Cell-culture-derived HCV (HCVcc) entry into hepatoma cell lines as well as primary human hepatocytes. The effect was independent of the HCV genotype, and both infection of cells by extracellular virions and cell-to-cell spread were blocked. Pretreatment of cells with EGCG before HCV inoculation did not reduce HCV infection, whereas the application of EGCG during inoculation strongly inhibited HCV infectivity. Moreover, treatment with EGCG directly during inoculation strongly inhibited HCV infectivity. Expression levels of all known HCV (co-)receptors were unaltered by EGCG. Finally, we showed that EGCG inhibits viral attachment to the cell, thus disrupting the initial step of HCV cell entry.
en
Catechin
Cell Line, Tumor
Cells, Cultured
Hepacivirus
Hepatitis C
Humans
Tea
Virus Attachment
Virus Internalization
The green tea polyphenol, epigallocatechin-3-gallate, inhibits hepatitis C virus entry.
Article2018-06-13T15:56:50ZHepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. Current antiviral therapy fails to clear infection in a substantial proportion of cases. Drug development is focused on nonstructural proteins required for RNA replication. Individuals undergoing orthotopic liver transplantation face rapid, universal reinfection of the graft. Therefore, antiviral strategies targeting the early stages of infection are urgently needed for the prevention of HCV infection. In this study, we identified the polyphenol, epigallocatechin-3-gallate (EGCG), as an inhibitor of HCV entry. Green tea catechins, such as EGCG and its derivatives, epigallocatechin (EGC), epicatechin gallate (ECG), and epicatechin (EC), have been previously found to exert antiviral and antioncogenic properties. EGCG had no effect on HCV RNA replication, assembly, or release of progeny virions. However, it potently inhibited Cell-culture-derived HCV (HCVcc) entry into hepatoma cell lines as well as primary human hepatocytes. The effect was independent of the HCV genotype, and both infection of cells by extracellular virions and cell-to-cell spread were blocked. Pretreatment of cells with EGCG before HCV inoculation did not reduce HCV infection, whereas the application of EGCG during inoculation strongly inhibited HCV infectivity. Moreover, treatment with EGCG directly during inoculation strongly inhibited HCV infectivity. Expression levels of all known HCV (co-)receptors were unaltered by EGCG. Finally, we showed that EGCG inhibits viral attachment to the cell, thus disrupting the initial step of HCV cell entry.oai:repository.helmholtz-hzi.de:10033/5504862019-08-30T11:25:11Zcom_10033_620589col_10033_620590
Bitzegeio, Julia
Bankwitz, Dorothea
Hueging, Kathrin
Haid, Sibylle
Brohm, Christiane
Zeisel, Mirjam B
Herrmann, Eva
Iken, Marcus
Ott, Michael
Baumert, Thomas F
Pietschmann, Thomas
2015-04-23T08:12:48Z
2015-04-23T08:12:48Z
2010
Adaptation of hepatitis C virus to mouse CD81 permits infection of mouse cells in the absence of human entry factors. 2010, 6:e1000978 PLoS Pathog.
1553-7374
20617177
10.1371/journal.ppat.1000978
http://hdl.handle.net/10033/550486
PLoS pathogens
Hepatitis C virus (HCV) naturally infects only humans and chimpanzees. The determinants responsible for this narrow species tropism are not well defined. Virus cell entry involves human scavenger receptor class B type I (SR-BI), CD81, claudin-1 and occludin. Among these, at least CD81 and occludin are utilized in a highly species-specific fashion, thus contributing to the narrow host range of HCV. We adapted HCV to mouse CD81 and identified three envelope glycoprotein mutations which together enhance infection of cells with mouse or other rodent receptors approximately 100-fold. These mutations enhanced interaction with human CD81 and increased exposure of the binding site for CD81 on the surface of virus particles. These changes were accompanied by augmented susceptibility of adapted HCV to neutralization by E2-specific antibodies indicative of major conformational changes of virus-resident E1/E2-complexes. Neutralization with CD81, SR-BI- and claudin-1-specific antibodies and knock down of occludin expression by siRNAs indicate that the adapted virus remains dependent on these host factors but apparently utilizes CD81, SR-BI and occludin with increased efficiency. Importantly, adapted E1/E2 complexes mediate HCV cell entry into mouse cells in the absence of human entry factors. These results further our knowledge of HCV receptor interactions and indicate that three glycoprotein mutations are sufficient to overcome the species-specific restriction of HCV cell entry into mouse cells. Moreover, these findings should contribute to the development of an immunocompetent small animal model fully permissive to HCV.
en
Animals
Antigens, CD
Antigens, CD81
Claudin-1
Hepacivirus
Membrane Proteins
Mice
Receptors, Virus
Scavenger Receptors, Class B
Viral Envelope Proteins
Virus Internalization
Adaptation of hepatitis C virus to mouse CD81 permits infection of mouse cells in the absence of human entry factors.
Article2018-06-13T00:18:00ZHepatitis C virus (HCV) naturally infects only humans and chimpanzees. The determinants responsible for this narrow species tropism are not well defined. Virus cell entry involves human scavenger receptor class B type I (SR-BI), CD81, claudin-1 and occludin. Among these, at least CD81 and occludin are utilized in a highly species-specific fashion, thus contributing to the narrow host range of HCV. We adapted HCV to mouse CD81 and identified three envelope glycoprotein mutations which together enhance infection of cells with mouse or other rodent receptors approximately 100-fold. These mutations enhanced interaction with human CD81 and increased exposure of the binding site for CD81 on the surface of virus particles. These changes were accompanied by augmented susceptibility of adapted HCV to neutralization by E2-specific antibodies indicative of major conformational changes of virus-resident E1/E2-complexes. Neutralization with CD81, SR-BI- and claudin-1-specific antibodies and knock down of occludin expression by siRNAs indicate that the adapted virus remains dependent on these host factors but apparently utilizes CD81, SR-BI and occludin with increased efficiency. Importantly, adapted E1/E2 complexes mediate HCV cell entry into mouse cells in the absence of human entry factors. These results further our knowledge of HCV receptor interactions and indicate that three glycoprotein mutations are sufficient to overcome the species-specific restriction of HCV cell entry into mouse cells. Moreover, these findings should contribute to the development of an immunocompetent small animal model fully permissive to HCV.oai:repository.helmholtz-hzi.de:10033/5523172019-08-30T11:25:11Zcom_10033_620589col_10033_620590
Steinmann, Eike
Pietschmann, Thomas
Division of Experimental Virology, Centre for Experimental and Clinical Infection Research, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2015-05-05T13:52:17Z
2015-05-05T13:52:17Z
2010-09
Hepatitis C virus p7-a viroporin crucial for virus assembly and an emerging target for antiviral therapy. 2010, 2 (9):2078-95 Viruses
1999-4915
21994720
10.3390/v2092078
http://hdl.handle.net/10033/552317
Viruses
The hepatitis C virus (HCV), a hepatotropic plus-strand RNA virus of the family Flaviviridae, encodes a set of 10 viral proteins. These viral factors act in concert with host proteins to mediate virus entry, and to coordinate RNA replication and virus production. Recent evidence has highlighted the complexity of HCV assembly, which not only involves viral structural proteins but also relies on host factors important for lipoprotein synthesis, and a number of viral assembly co-factors. The latter include the integral membrane protein p7, which oligomerizes and forms cation-selective pores. Based on these properties, p7 was included into the family of viroporins comprising viral proteins from multiple virus families which share the ability to manipulate membrane permeability for ions and to facilitate virus production. Although the precise mechanism as to how p7 and its ion channel function contributes to virus production is still elusive, recent structural and functional studies have revealed a number of intriguing new facets that should guide future efforts to dissect the role and function of p7 in the viral replication cycle. Moreover, a number of small molecules that inhibit production of HCV particles, presumably via interference with p7 function, have been reported. These compounds should not only be instrumental in increasing our understanding of p7 function, but may, in the future, merit further clinical development to ultimately optimize HCV-specific antiviral treatments.
en
Hepatitis C virus p7-a viroporin crucial for virus assembly and an emerging target for antiviral therapy.
Article2018-06-12T21:20:51ZThe hepatitis C virus (HCV), a hepatotropic plus-strand RNA virus of the family Flaviviridae, encodes a set of 10 viral proteins. These viral factors act in concert with host proteins to mediate virus entry, and to coordinate RNA replication and virus production. Recent evidence has highlighted the complexity of HCV assembly, which not only involves viral structural proteins but also relies on host factors important for lipoprotein synthesis, and a number of viral assembly co-factors. The latter include the integral membrane protein p7, which oligomerizes and forms cation-selective pores. Based on these properties, p7 was included into the family of viroporins comprising viral proteins from multiple virus families which share the ability to manipulate membrane permeability for ions and to facilitate virus production. Although the precise mechanism as to how p7 and its ion channel function contributes to virus production is still elusive, recent structural and functional studies have revealed a number of intriguing new facets that should guide future efforts to dissect the role and function of p7 in the viral replication cycle. Moreover, a number of small molecules that inhibit production of HCV particles, presumably via interference with p7 function, have been reported. These compounds should not only be instrumental in increasing our understanding of p7 function, but may, in the future, merit further clinical development to ultimately optimize HCV-specific antiviral treatments.oai:repository.helmholtz-hzi.de:10033/5541782019-08-30T11:33:29Zcom_10033_620589col_10033_620590
Anggakusuma
Frentzen, Anne
Gürlevik, Engin
Yuan, Qinggong
Steinmann, Eike
Ott, Michael
Staeheli, Peter
Schmid-Burgk, Jonathan
Schmidt, Tobias
Hornung, Veit
Kuehnel, Florian
Pietschmann, Thomas
Institute of Experimental Virology, Twincore Centre for Experimental and Clinical Infection Research, Hanover, Germany.
2015-05-19T15:19:36Z
2015-05-19T15:19:36Z
2015-04
Control of hepatitis C virus replication in mouse liver-derived cells by MAVS-dependent production of type I and type III interferons. 2015, 89 (7):3833-45 J. Virol.
1098-5514
25609814
10.1128/JVI.03129-14
http://hdl.handle.net/10033/554178
Journal of virology
Hepatitis C virus (HCV) efficiently infects only humans and chimpanzees. Although the detailed mechanisms responsible for this narrow species tropism remain elusive, recent evidence has shown that murine innate immune responses efficiently suppress HCV replication. Therefore, poor adaptation of HCV to evade and/or counteract innate immune responses may prevent HCV replication in mice. The HCV NS3-4A protease cleaves human MAVS, a key cellular adaptor protein required for RIG-I-like receptor (RLR)-dependent innate immune signaling. However, it is unclear if HCV interferes with mouse MAVS function equally well. Moreover, MAVS-dependent signaling events that restrict HCV replication in mouse cells were incompletely defined. Thus, we quantified the ability of HCV NS3-4A to counteract mouse and human MAVS. HCV NS3-4A similarly diminished both human and mouse MAVS-dependent signaling in human and mouse cells. Moreover, replicon-encoded protease cleaved a similar fraction of both MAVS variants. Finally, FLAG-tagged MAVS proteins repressed HCV replication to similar degrees. Depending on MAVS expression, HCV replication in mouse liver cells triggered not only type I but also type III IFNs, which cooperatively repressed HCV replication. Mouse liver cells lacking both type I and III IFN receptors were refractory to MAVS-dependent antiviral effects, indicating that the HCV-induced MAVS-dependent antiviral state depends on both type I and III IFN receptor signaling.
en
Control of hepatitis C virus replication in mouse liver-derived cells by MAVS-dependent production of type I and type III interferons.
Article2018-06-13T00:29:08ZHepatitis C virus (HCV) efficiently infects only humans and chimpanzees. Although the detailed mechanisms responsible for this narrow species tropism remain elusive, recent evidence has shown that murine innate immune responses efficiently suppress HCV replication. Therefore, poor adaptation of HCV to evade and/or counteract innate immune responses may prevent HCV replication in mice. The HCV NS3-4A protease cleaves human MAVS, a key cellular adaptor protein required for RIG-I-like receptor (RLR)-dependent innate immune signaling. However, it is unclear if HCV interferes with mouse MAVS function equally well. Moreover, MAVS-dependent signaling events that restrict HCV replication in mouse cells were incompletely defined. Thus, we quantified the ability of HCV NS3-4A to counteract mouse and human MAVS. HCV NS3-4A similarly diminished both human and mouse MAVS-dependent signaling in human and mouse cells. Moreover, replicon-encoded protease cleaved a similar fraction of both MAVS variants. Finally, FLAG-tagged MAVS proteins repressed HCV replication to similar degrees. Depending on MAVS expression, HCV replication in mouse liver cells triggered not only type I but also type III IFNs, which cooperatively repressed HCV replication. Mouse liver cells lacking both type I and III IFN receptors were refractory to MAVS-dependent antiviral effects, indicating that the HCV-induced MAVS-dependent antiviral state depends on both type I and III IFN receptor signaling.oai:repository.helmholtz-hzi.de:10033/5795742019-08-30T11:34:48Zcom_10033_620589col_10033_620590
Lump, Edina
Castellano, Laura M
Meier, Christoph
Seeliger, Janine
Erwin, Nelli
Sperlich, Benjamin
Stürzel, Christina M
Usmani, Shariq
Hammond, Rebecca M
von Einem, Jens
Gerold, Gisa
Kreppel, Florian
Bravo-Rodriguez, Kenny
Pietschmann, Thomas
Holmes, Veronica M
Palesch, David
Zirafi, Onofrio
Weissman, Drew
Sowislok, Andrea
Wettig, Burkhard
Heid, Christian
Kirchhoff, Frank
Weil, Tanja
Klärner, Frank-Gerrit
Schrader, Thomas
Bitan, Gal
Sanchez-Garcia, Elsa
Winter, Roland
Shorter, James
Münch, Jan
Institute of Experimental Virology, Twincore, Centre for Experimental and Clinical Infection Research, Hannover, Germany.
2015-10-12T13:38:58Z
2015-10-12T13:38:58Z
2015
A molecular tweezer antagonizes seminal amyloids and HIV infection. 2015, 4: Elife
2050-084X
26284498
10.7554/eLife.05397
http://hdl.handle.net/10033/579574
eLife
Semen is the main vector for HIV transmission and contains amyloid fibrils that enhance viral infection. Available microbicides that target viral components have proven largely ineffective in preventing sexual virus transmission. In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils. Moreover, CLR01 abrogates semen-mediated enhancement of viral infection by preventing the formation of virion-amyloid complexes and by directly disrupting the membrane integrity of HIV and other enveloped viruses. We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism. CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself. These combined anti-amyloid and antiviral activities make CLR01 a promising topical microbicide for blocking infection by HIV and other sexually transmitted viruses.
en
A molecular tweezer antagonizes seminal amyloids and HIV infection.
Article2018-06-13T16:00:54ZSemen is the main vector for HIV transmission and contains amyloid fibrils that enhance viral infection. Available microbicides that target viral components have proven largely ineffective in preventing sexual virus transmission. In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils. Moreover, CLR01 abrogates semen-mediated enhancement of viral infection by preventing the formation of virion-amyloid complexes and by directly disrupting the membrane integrity of HIV and other enveloped viruses. We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism. CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself. These combined anti-amyloid and antiviral activities make CLR01 a promising topical microbicide for blocking infection by HIV and other sexually transmitted viruses.oai:repository.helmholtz-hzi.de:10033/5932842019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Nadeem, Abd-Elshafy D
Thomas, Pietschmann
Ulf, Müller-Ladner
Elena, Neumann
Anggakusuma
Mohamed, Bahgat M
Frank, Pessler
Patrick, Behrendt
TWINCORE, Centre for Experimental and Clinical Infection Research GmbH, Feodor-Lynen-Str. 3-7, 30625 Hannover, Germany.
2016-01-11T15:44:25Z
2016-01-11T15:44:25Z
2015
Cell culture-derived HCV cannot infect synovial fibroblasts. 2015, 5:18043 Sci Rep
2045-2322
26643193
10.1038/srep18043
http://hdl.handle.net/10033/593284
Scientific reports
Worldwide 170 million individuals are infected with hepatitis C virus (HCV), up to 45 million of whom are affected by arthropathy. It is unclear whether this is due to viral infection of synovial cells or immune-mediated mechanisms. We tested the capacity of primary synovial fibroblasts to support HCV propagation. Out of the four critical HCV receptors, only CD81 was expressed to any significant extent in OASF and RASF. Consistent with this, pseudotyped HCV particles were unable to infect these cells. Permissiveness for HCV replication was investigated by transfecting cells with a subgenomic replicon of HCV encoding a luciferase reporter. OASF and RASF did not support replication of HCV, possibly due to low expression levels of miR-122. In conclusion, primary human synovial fibroblasts are unable to support propagation of HCV in vitro. HCV-related arthropathy is unlikely due to direct infection of these cells.
en
Cell culture-derived HCV cannot infect synovial fibroblasts.
Article2018-06-12T22:07:51ZWorldwide 170 million individuals are infected with hepatitis C virus (HCV), up to 45 million of whom are affected by arthropathy. It is unclear whether this is due to viral infection of synovial cells or immune-mediated mechanisms. We tested the capacity of primary synovial fibroblasts to support HCV propagation. Out of the four critical HCV receptors, only CD81 was expressed to any significant extent in OASF and RASF. Consistent with this, pseudotyped HCV particles were unable to infect these cells. Permissiveness for HCV replication was investigated by transfecting cells with a subgenomic replicon of HCV encoding a luciferase reporter. OASF and RASF did not support replication of HCV, possibly due to low expression levels of miR-122. In conclusion, primary human synovial fibroblasts are unable to support propagation of HCV in vitro. HCV-related arthropathy is unlikely due to direct infection of these cells.oai:repository.helmholtz-hzi.de:10033/5963442019-08-30T11:35:14Zcom_10033_620589col_10033_620590
Perin, Paula M
Haid, Sibylle
Brown, Richard J P
Doerrbecker, Juliane
Schulze, Kai
Zeilinger, Carsten
von Schaewen, Markus
Heller, Brigitte
Vercauteren, Koen
Luxenburger, Eva
Baktash, Yasmine M
Vondran, Florian W R
Speerstra, Sietkse
Awadh, Abdullah
Mukhtarov, Furkat
Schang, Luis M
Kirschning, Andreas
Müller, Rolf
Guzman, Carlos A
Kaderali, Lars
Randall, Glenn
Meuleman, Philip
Ploss, Alexander
Thomas, Pietschmann
TWINCORE, Centre for Experimental and Clinical Infection Research GmbH, Feodor-Lynen-Str. 3-7, 30625 Hannover, Germany.
2016-02-16T14:38:17Z
2016-02-16T14:38:17Z
2016-01
Flunarizine prevents hepatitis C virus membrane fusion in a genotype-dependent manner by targeting the potential fusion peptide within E1. 2016, 63 (1):49-62 Hepatology
1527-3350
26248546
10.1002/hep.28111
http://hdl.handle.net/10033/596344
Hepatology (Baltimore, Md.)
To explore mechanisms of hepatitis C viral (HCV) replication we screened a compound library including licensed drugs. Flunarizine, a diphenylmethylpiperazine used to treat migraine, inhibited HCV cell entry in vitro and in vivo in a genotype-dependent fashion. Analysis of mosaic viruses between susceptible and resistant strains revealed that E1 and E2 glycoproteins confer susceptibility to flunarizine. Time of addition experiments and single particle tracking of HCV demonstrated that flunarizine specifically prevents membrane fusion. Related phenothiazines and pimozide also inhibited HCV infection and preferentially targeted HCV genotype 2 viruses. However, phenothiazines and pimozide exhibited improved genotype coverage including the difficult to treat genotype 3. Flunarizine-resistant HCV carried mutations within the alleged fusion peptide and displayed cross-resistance to these compounds, indicating that these drugs have a common mode of action.
en
Flunarizine prevents hepatitis C virus membrane fusion in a genotype-dependent manner by targeting the potential fusion peptide within E1.
Article2017-01-01T00:00:00ZTo explore mechanisms of hepatitis C viral (HCV) replication we screened a compound library including licensed drugs. Flunarizine, a diphenylmethylpiperazine used to treat migraine, inhibited HCV cell entry in vitro and in vivo in a genotype-dependent fashion. Analysis of mosaic viruses between susceptible and resistant strains revealed that E1 and E2 glycoproteins confer susceptibility to flunarizine. Time of addition experiments and single particle tracking of HCV demonstrated that flunarizine specifically prevents membrane fusion. Related phenothiazines and pimozide also inhibited HCV infection and preferentially targeted HCV genotype 2 viruses. However, phenothiazines and pimozide exhibited improved genotype coverage including the difficult to treat genotype 3. Flunarizine-resistant HCV carried mutations within the alleged fusion peptide and displayed cross-resistance to these compounds, indicating that these drugs have a common mode of action.oai:repository.helmholtz-hzi.de:10033/6012232019-08-30T11:35:13Zcom_10033_620589col_10033_620590
Walker, Andreas
Skibbe, Kathrin
Steinmann, Eike
Pfaender, Stephanie
Kuntzen, Thomas
Megger, Dominik A
Groten, Svenja
Sitek, Barbara
Lauer, Georg M
Kim, Arthur Y
Pietschmann, Thomas
Allen, Todd M
Timm, Joerg
TWINCORE Centre for Experimental and Clinical Infection Research, Feodor-Lynen-Str. 7, 30625, Hannover, Germany.
2016-03-11T15:24:56Z
2016-03-11T15:24:56Z
2015
Distinct Escape Pathway by Hepatitis C Virus Genotype 1a from a Dominant CD8+ T Cell Response by Selection of Altered Epitope Processing. 2015, 90 (1):33-42 J. Virol.
1098-5514
26446603
10.1128/JVI.01993-15
http://hdl.handle.net/10033/601223
Journal of virology
Antiviral CD8(+) T cells are a key component of the adaptive immune response against HCV, but their impact on viral control is influenced by preexisting viral variants in important target epitopes and the development of viral escape mutations. Immunodominant epitopes highly conserved across genotypes therefore are attractive for T cell based prophylactic vaccines. Here, we characterized the CD8(+) T cell response against the highly conserved HLA-B*51-restricted epitope IPFYGKAI1373-1380 located in the helicase domain of NS3 in people who inject drugs (PWID) exposed predominantly to HCV genotypes 1a and 3a. Despite this epitope being conserved in both genotypes, the corresponding CD8(+) T cell response was detected only in PWID infected with genotype 3a and HCV-RNA negative PWID, but not in PWID infected with genotype 1a. In genotype 3a, the detection of strong CD8(+) T cell responses was associated with epitope variants in the autologous virus consistent with immune escape. Analysis of viral sequences from multiple cohorts confirmed HLA-B*51-associated escape mutations inside the epitope in genotype 3a, but not in genotype 1a. Here, a distinct substitution in the N-terminal flanking region located 5 residues upstream of the epitope (S1368P; P = 0.00002) was selected in HLA-B*51-positive individuals. Functional assays revealed that the S1368P substitution impaired recognition of target cells presenting the endogenously processed epitope. The results highlight that, despite an epitope being highly conserved between two genotypes, there are major differences in the selected viral escape pathways and the corresponding T cell responses.
en
Distinct Escape Pathway by Hepatitis C Virus Genotype 1a from a Dominant CD8+ T Cell Response by Selection of Altered Epitope Processing.
Article2016-07-15T00:00:00ZAntiviral CD8(+) T cells are a key component of the adaptive immune response against HCV, but their impact on viral control is influenced by preexisting viral variants in important target epitopes and the development of viral escape mutations. Immunodominant epitopes highly conserved across genotypes therefore are attractive for T cell based prophylactic vaccines. Here, we characterized the CD8(+) T cell response against the highly conserved HLA-B*51-restricted epitope IPFYGKAI1373-1380 located in the helicase domain of NS3 in people who inject drugs (PWID) exposed predominantly to HCV genotypes 1a and 3a. Despite this epitope being conserved in both genotypes, the corresponding CD8(+) T cell response was detected only in PWID infected with genotype 3a and HCV-RNA negative PWID, but not in PWID infected with genotype 1a. In genotype 3a, the detection of strong CD8(+) T cell responses was associated with epitope variants in the autologous virus consistent with immune escape. Analysis of viral sequences from multiple cohorts confirmed HLA-B*51-associated escape mutations inside the epitope in genotype 3a, but not in genotype 1a. Here, a distinct substitution in the N-terminal flanking region located 5 residues upstream of the epitope (S1368P; P = 0.00002) was selected in HLA-B*51-positive individuals. Functional assays revealed that the S1368P substitution impaired recognition of target cells presenting the endogenously processed epitope. The results highlight that, despite an epitope being highly conserved between two genotypes, there are major differences in the selected viral escape pathways and the corresponding T cell responses.oai:repository.helmholtz-hzi.de:10033/6062562019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Walker, Andreas
Skibbe, Kathrin
Steinmann, Eike
Pfaender, Stephanie
Kuntzen, Thomas
Megger, Dominik A
Groten, Svenja
Sitek, Barbara
Lauer, Georg M
Kim, Arthur Y
Pietschmann, Thomas
Allen, Todd M
Timm, Joerg
Twincore Centre for Experimental and Clinical Infection Research, a joint venture between the Medical School Hanover and the Helmholtz Centre for Infection Research, Hanover, Germany.
2016-04-21T08:58:39Z
2016-04-21T08:58:39Z
2015
Distinct Escape Pathway by Hepatitis C Virus Genotype 1a from a Dominant CD8+ T Cell Response by Selection of Altered Epitope Processing. 2015, 90 (1):33-42 J. Virol.
1098-5514
26446603
10.1128/JVI.01993-15
http://hdl.handle.net/10033/606256
Journal of virology
Antiviral CD8(+) T cells are a key component of the adaptive immune response against HCV, but their impact on viral control is influenced by preexisting viral variants in important target epitopes and the development of viral escape mutations. Immunodominant epitopes highly conserved across genotypes therefore are attractive for T cell based prophylactic vaccines. Here, we characterized the CD8(+) T cell response against the highly conserved HLA-B*51-restricted epitope IPFYGKAI1373-1380 located in the helicase domain of NS3 in people who inject drugs (PWID) exposed predominantly to HCV genotypes 1a and 3a. Despite this epitope being conserved in both genotypes, the corresponding CD8(+) T cell response was detected only in PWID infected with genotype 3a and HCV-RNA negative PWID, but not in PWID infected with genotype 1a. In genotype 3a, the detection of strong CD8(+) T cell responses was associated with epitope variants in the autologous virus consistent with immune escape. Analysis of viral sequences from multiple cohorts confirmed HLA-B*51-associated escape mutations inside the epitope in genotype 3a, but not in genotype 1a. Here, a distinct substitution in the N-terminal flanking region located 5 residues upstream of the epitope (S1368P; P = 0.00002) was selected in HLA-B*51-positive individuals. Functional assays revealed that the S1368P substitution impaired recognition of target cells presenting the endogenously processed epitope. The results highlight that, despite an epitope being highly conserved between two genotypes, there are major differences in the selected viral escape pathways and the corresponding T cell responses.
en
Distinct Escape Pathway by Hepatitis C Virus Genotype 1a from a Dominant CD8+ T Cell Response by Selection of Altered Epitope Processing.
Article2018-06-13T00:00:46ZAntiviral CD8(+) T cells are a key component of the adaptive immune response against HCV, but their impact on viral control is influenced by preexisting viral variants in important target epitopes and the development of viral escape mutations. Immunodominant epitopes highly conserved across genotypes therefore are attractive for T cell based prophylactic vaccines. Here, we characterized the CD8(+) T cell response against the highly conserved HLA-B*51-restricted epitope IPFYGKAI1373-1380 located in the helicase domain of NS3 in people who inject drugs (PWID) exposed predominantly to HCV genotypes 1a and 3a. Despite this epitope being conserved in both genotypes, the corresponding CD8(+) T cell response was detected only in PWID infected with genotype 3a and HCV-RNA negative PWID, but not in PWID infected with genotype 1a. In genotype 3a, the detection of strong CD8(+) T cell responses was associated with epitope variants in the autologous virus consistent with immune escape. Analysis of viral sequences from multiple cohorts confirmed HLA-B*51-associated escape mutations inside the epitope in genotype 3a, but not in genotype 1a. Here, a distinct substitution in the N-terminal flanking region located 5 residues upstream of the epitope (S1368P; P = 0.00002) was selected in HLA-B*51-positive individuals. Functional assays revealed that the S1368P substitution impaired recognition of target cells presenting the endogenously processed epitope. The results highlight that, despite an epitope being highly conserved between two genotypes, there are major differences in the selected viral escape pathways and the corresponding T cell responses.oai:repository.helmholtz-hzi.de:10033/6051022019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Urbanowicz, Richard A
McClure, C Patrick
Brown, Richard J P
Tsoleridis, Theocharis
Persson, Mats A A
Krey, Thomas
Irving, William L
Ball, Jonathan K
Tarr, Alexander W
Twincore Centre of Experimental and Clinical Infection Research; a joint venture between the Hannover Medical School and the Helmholtz Centre for Infection Research, Hannover 30625, Germany.
2016-04-12T14:35:11Z
2016-04-12T14:35:11Z
2015
A Diverse Panel of Hepatitis C Virus Glycoproteins for Use in Vaccine Research Reveals Extremes of Monoclonal Antibody Neutralization Resistance. 2015, 90 (7):3288-301 J. Virol.
1098-5514
26699643
10.1128/JVI.02700-15
http://hdl.handle.net/10033/605102
Journal of virology
Despite significant advances in the treatment of hepatitis C virus (HCV) infection, the need to develop preventative vaccines remains. Identification of the best vaccine candidates and evaluation of their performance in preclinical and clinical development will require appropriate neutralization assays utilizing diverse HCV isolates. We aimed to generate and characterize a panel of HCV E1E2 glycoproteins suitable for subsequent use in vaccine and therapeutic antibody testing. Full-length E1E2 clones were PCR amplified from patient-derived serum samples, cloned into an expression vector, and used to generate viral pseudoparticles (HCVpp). In addition, some of these clones were used to generate cell culture infectious (HCVcc) clones. The infectivity and neutralization sensitivity of these viruses were then determined. Bioinformatic and HCVpp infectivity screening of approximately 900 E1E2 clones resulted in the assembly of a panel of 78 functional E1E2 proteins representing distinct HCV genotypes and different stages of infection. These HCV glycoproteins differed markedly in their sensitivity to neutralizing antibodies. We used this panel to predict antibody efficacy against circulating HCV strains, highlighting the likely reason why some monoclonal antibodies failed in previous clinical trials. This study provides the first objective categorization of cross-genotype patient-derived HCV E1E2 clones according to their sensitivity to antibody neutralization. It has shown that HCV isolates have clearly distinguishable neutralization-sensitive, -resistant, or -intermediate phenotypes, which are independent of genotype. The panel provides a systematic means for characterization of the neutralizing response elicited by candidate vaccines and for defining the therapeutic potential of monoclonal antibodies.
en
A Diverse Panel of Hepatitis C Virus Glycoproteins for Use in Vaccine Research Reveals Extremes of Monoclonal Antibody Neutralization Resistance.
Article2018-06-14T09:18:36ZDespite significant advances in the treatment of hepatitis C virus (HCV) infection, the need to develop preventative vaccines remains. Identification of the best vaccine candidates and evaluation of their performance in preclinical and clinical development will require appropriate neutralization assays utilizing diverse HCV isolates. We aimed to generate and characterize a panel of HCV E1E2 glycoproteins suitable for subsequent use in vaccine and therapeutic antibody testing. Full-length E1E2 clones were PCR amplified from patient-derived serum samples, cloned into an expression vector, and used to generate viral pseudoparticles (HCVpp). In addition, some of these clones were used to generate cell culture infectious (HCVcc) clones. The infectivity and neutralization sensitivity of these viruses were then determined. Bioinformatic and HCVpp infectivity screening of approximately 900 E1E2 clones resulted in the assembly of a panel of 78 functional E1E2 proteins representing distinct HCV genotypes and different stages of infection. These HCV glycoproteins differed markedly in their sensitivity to neutralizing antibodies. We used this panel to predict antibody efficacy against circulating HCV strains, highlighting the likely reason why some monoclonal antibodies failed in previous clinical trials. This study provides the first objective categorization of cross-genotype patient-derived HCV E1E2 clones according to their sensitivity to antibody neutralization. It has shown that HCV isolates have clearly distinguishable neutralization-sensitive, -resistant, or -intermediate phenotypes, which are independent of genotype. The panel provides a systematic means for characterization of the neutralizing response elicited by candidate vaccines and for defining the therapeutic potential of monoclonal antibodies.oai:repository.helmholtz-hzi.de:10033/6077362019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Vieyres, Gabrielle
Welsch, Kathrin
Gerold, Gisa
Gentzsch, Juliane
Kahl, Sina
Vondran, Florian W R
Kaderali, Lars
Pietschmann, Thomas
Twincore Centre for Experimental and Clinical Infection Research, Hannover, Germany.
2016-05-03T13:15:16Z
2016-05-03T13:15:16Z
2016-04
ABHD5/CGI-58, the Chanarin-Dorfman Syndrome Protein, Mobilises Lipid Stores for Hepatitis C Virus Production. 2016, 12 (4):e1005568 PLoS Pathog.
1553-7374
27124600
10.1371/journal.ppat.1005568
http://hdl.handle.net/10033/607736
PLoS pathogens
Hepatitis C virus (HCV) particles closely mimic human very-low-density lipoproteins (VLDL) to evade humoral immunity and to facilitate cell entry. However, the principles that govern HCV association with VLDL components are poorly defined. Using an siRNA screen, we identified ABHD5 (α/β hydrolase domain containing protein 5, also known as CGI-58) as a new host factor promoting both virus assembly and release. ABHD5 associated with lipid droplets and triggered their hydrolysis. Importantly, ABHD5 Chanarin-Dorfman syndrome mutants responsible for a rare lipid storage disorder in humans were mislocalised, and unable to consume lipid droplets or support HCV production. Additional ABHD5 mutagenesis revealed a novel tribasic motif that does not influence subcellular localization but determines both ABHD5 lipolytic and proviral properties. These results indicate that HCV taps into the lipid droplet triglyceride reservoir usurping ABHD5 lipase cofactor function. They also suggest that the resulting lipid flux, normally devoted to VLDL synthesis, also participates in the assembly and release of the HCV lipo-viro-particle. Altogether, our study provides the first association between the Chanarin-Dorfman syndrome protein and an infectious disease and sheds light on the hepatic manifestations of this rare genetic disorder as well as on HCV morphogenesis.
en
ABHD5/CGI-58, the Chanarin-Dorfman Syndrome Protein, Mobilises Lipid Stores for Hepatitis C Virus Production.
Article2018-06-13T00:52:27ZHepatitis C virus (HCV) particles closely mimic human very-low-density lipoproteins (VLDL) to evade humoral immunity and to facilitate cell entry. However, the principles that govern HCV association with VLDL components are poorly defined. Using an siRNA screen, we identified ABHD5 (α/β hydrolase domain containing protein 5, also known as CGI-58) as a new host factor promoting both virus assembly and release. ABHD5 associated with lipid droplets and triggered their hydrolysis. Importantly, ABHD5 Chanarin-Dorfman syndrome mutants responsible for a rare lipid storage disorder in humans were mislocalised, and unable to consume lipid droplets or support HCV production. Additional ABHD5 mutagenesis revealed a novel tribasic motif that does not influence subcellular localization but determines both ABHD5 lipolytic and proviral properties. These results indicate that HCV taps into the lipid droplet triglyceride reservoir usurping ABHD5 lipase cofactor function. They also suggest that the resulting lipid flux, normally devoted to VLDL synthesis, also participates in the assembly and release of the HCV lipo-viro-particle. Altogether, our study provides the first association between the Chanarin-Dorfman syndrome protein and an infectious disease and sheds light on the hepatic manifestations of this rare genetic disorder as well as on HCV morphogenesis.oai:repository.helmholtz-hzi.de:10033/6090552019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Carpentier, Arnaud
Nimgaonkar, Ila
Chu, Virginia
Xia, Yuchen
Hu, Zongyi
Liang, T Jake
Twincore, Institute for Experimental Virology, Hannover, Germany.
2016-05-11T14:20:35Z
2016-05-11T14:20:35Z
2016-03-29
Hepatic differentiation of human pluripotent stem cells in miniaturized format suitable for high-throughput screen. 2016, 16 (3):640-650 Stem Cell Res
1876-7753
27062358
10.1016/j.scr.2016.03.009
http://hdl.handle.net/10033/609055
Stem cell research
The establishment of protocols to differentiate human pluripotent stem cells (hPSCs) including embryonic (ESC) and induced pluripotent (iPSC) stem cells into functional hepatocyte-like cells (HLCs) creates new opportunities to study liver metabolism, genetic diseases and infection of hepatotropic viruses (hepatitis B and C viruses) in the context of specific genetic background. While supporting efficient differentiation to HLCs, the published protocols are limited in terms of differentiation into fully mature hepatocytes and in a smaller-well format. This limitation handicaps the application of these cells to high-throughput assays. Here we describe a protocol allowing efficient and consistent hepatic differentiation of hPSCs in 384-well plates into functional hepatocyte-like cells, which remain differentiated for more than 3weeks. This protocol affords the unique opportunity to miniaturize the hPSC-based differentiation technology and facilitates screening for molecules in modulating liver differentiation, metabolism, genetic network, and response to infection or other external stimuli.
ENG
Hepatic differentiation of human pluripotent stem cells in miniaturized format suitable for high-throughput screen.
Article2018-06-13T00:07:08ZThe establishment of protocols to differentiate human pluripotent stem cells (hPSCs) including embryonic (ESC) and induced pluripotent (iPSC) stem cells into functional hepatocyte-like cells (HLCs) creates new opportunities to study liver metabolism, genetic diseases and infection of hepatotropic viruses (hepatitis B and C viruses) in the context of specific genetic background. While supporting efficient differentiation to HLCs, the published protocols are limited in terms of differentiation into fully mature hepatocytes and in a smaller-well format. This limitation handicaps the application of these cells to high-throughput assays. Here we describe a protocol allowing efficient and consistent hepatic differentiation of hPSCs in 384-well plates into functional hepatocyte-like cells, which remain differentiated for more than 3weeks. This protocol affords the unique opportunity to miniaturize the hPSC-based differentiation technology and facilitates screening for molecules in modulating liver differentiation, metabolism, genetic network, and response to infection or other external stimuli.oai:repository.helmholtz-hzi.de:10033/6116972019-08-30T11:33:30Zcom_10033_620589col_10033_620590
Hueging, Kathrin
Weller, Romy
Doepke, Mandy
Vieyres, Gabrielle
Todt, Daniel
Wölk, Benno
Vondran, Florian W R
Geffers, Robert
Lauber, Chris
Kaderali, Lars
Penin, François
Pietschmann, Thomas
Twincore Centre of Experimental and Clinical Infection Research; a joint venture between the Hannover Medical School and the Helmholtz Centre for Infection Research, Hannover 30625, Germany.
2016-06-03T09:22:58Z
2016-06-03T09:22:58Z
2015
Several Human Liver Cell Expressed Apolipoproteins Complement HCV Virus Production with Varying Efficacy Conferring Differential Specific Infectivity to Released Viruses. 2015, 10 (7):e0134529 PLoS ONE
1932-6203
26226615
10.1371/journal.pone.0134529
http://hdl.handle.net/10033/611697
PloS one
Apolipoprotein E (ApoE), an exchangeable apolipoprotein, is necessary for production of infectious Hepatitis C virus (HCV) particles. However, ApoE is not the only liver-expressed apolipoprotein and the role of other apolipoproteins for production of infectious HCV progeny is incompletely defined. Therefore, we quantified mRNA expression of human apolipoproteins in primary human hepatocytes. Subsequently, cDNAs encoding apolipoproteins were expressed in 293T/miR-122 cells to explore if they complement HCV virus production in cells that are non-permissive due to limiting endogenous levels of human apolipoproteins. Primary human hepatocytes expressed high mRNA levels of ApoA1, A2, C1, C3, E, and H. ApoA4, A5, B, D, F, J, L1, L2, L3, L4, L6, M, and O were expressed at intermediate levels, and C2, C4, and L5 were not detected. All members of the ApoA and ApoC family of lipoproteins complemented HCV virus production in HCV transfected 293T/miR-122 cells, albeit with significantly lower efficacy compared with ApoE. In contrast, ApoD expression did not support production of infectious HCV. Specific infectivity of released particles complemented with ApoA family members was significantly lower compared with ApoE. Moreover, the ratio of extracellular to intracellular infectious virus was significantly higher for ApoE compared to ApoA2 and ApoC3. Since apolipoproteins complementing HCV virus production share amphipathic alpha helices as common structural features we altered the two alpha helices of ApoC1. Helix breaking mutations in both ApoC1 helices impaired virus assembly highlighting a critical role of alpha helices in apolipoproteins supporting HCV assembly. In summary, various liver expressed apolipoproteins with amphipathic alpha helices complement HCV virus production in human non liver cells. Differences in the efficiency of virus assembly, the specific infectivity of released particles, and the ratio between extracellular and intracellular infectivity point to distinct characteristics of these apolipoproteins that influence HCV assembly and cell entry. This will guide future research to precisely pinpoint how apolipoproteins function during virus assembly and cell entry.
en
Apolipoproteins
Cell Line
Hepacivirus
Hepatocytes
Humans
Virulence
Virus Replication
Several Human Liver Cell Expressed Apolipoproteins Complement HCV Virus Production with Varying Efficacy Conferring Differential Specific Infectivity to Released Viruses.
Article2018-06-13T00:12:50ZApolipoprotein E (ApoE), an exchangeable apolipoprotein, is necessary for production of infectious Hepatitis C virus (HCV) particles. However, ApoE is not the only liver-expressed apolipoprotein and the role of other apolipoproteins for production of infectious HCV progeny is incompletely defined. Therefore, we quantified mRNA expression of human apolipoproteins in primary human hepatocytes. Subsequently, cDNAs encoding apolipoproteins were expressed in 293T/miR-122 cells to explore if they complement HCV virus production in cells that are non-permissive due to limiting endogenous levels of human apolipoproteins. Primary human hepatocytes expressed high mRNA levels of ApoA1, A2, C1, C3, E, and H. ApoA4, A5, B, D, F, J, L1, L2, L3, L4, L6, M, and O were expressed at intermediate levels, and C2, C4, and L5 were not detected. All members of the ApoA and ApoC family of lipoproteins complemented HCV virus production in HCV transfected 293T/miR-122 cells, albeit with significantly lower efficacy compared with ApoE. In contrast, ApoD expression did not support production of infectious HCV. Specific infectivity of released particles complemented with ApoA family members was significantly lower compared with ApoE. Moreover, the ratio of extracellular to intracellular infectious virus was significantly higher for ApoE compared to ApoA2 and ApoC3. Since apolipoproteins complementing HCV virus production share amphipathic alpha helices as common structural features we altered the two alpha helices of ApoC1. Helix breaking mutations in both ApoC1 helices impaired virus assembly highlighting a critical role of alpha helices in apolipoproteins supporting HCV assembly. In summary, various liver expressed apolipoproteins with amphipathic alpha helices complement HCV virus production in human non liver cells. Differences in the efficiency of virus assembly, the specific infectivity of released particles, and the ratio between extracellular and intracellular infectivity point to distinct characteristics of these apolipoproteins that influence HCV assembly and cell entry. This will guide future research to precisely pinpoint how apolipoproteins function during virus assembly and cell entry.oai:repository.helmholtz-hzi.de:10033/6158132019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Gerold, Gisa
Bruening, Janina
Pietschmann, Thomas
TWINCORE, Centre for Experimental and Clinical Infection Research, Feodor-Lynen-Str. 7 30625 Hannover, Germany.
2016-07-08T14:31:39Z
2016-07-08T14:31:39Z
2016-06-15
Decoding protein networks during virus entry by quantitative proteomics. 2016, 218:25-39 Virus Res.
1872-7492
26365680
10.1016/j.virusres.2015.09.006
http://hdl.handle.net/10033/615813
Virus research
Virus entry into host cells relies on interactions between viral and host structures including lipids, carbohydrates and proteins. Particularly, protein-protein interactions between viral surface proteins and host proteins as well as secondary host protein-protein interactions play a pivotal role in coordinating virus binding and uptake. These interactions are dynamic and frequently involve multiprotein complexes. In the past decade mass spectrometry based proteomics methods have reached sensitivities and high throughput compatibilities of genomics methods and now allow the reliable quantitation of proteins in complex samples from limited material. As proteomics provides essential information on the biologically active entity namely the protein, including its posttranslational modifications and its interactions with other proteins, it is an indispensable method in the virologist's toolbox. Here we review protein interactions during virus entry and compare classical biochemical methods to study entry with novel technically advanced quantitative proteomics techniques. We highlight the value of quantitative proteomics in mapping functional virus entry networks, discuss the benefits and limitations and illustrate how the methodology will help resolve unsettled questions in virus entry research in the future.
en
info:eu-repo/grantAgreement/EC/FP7/281473
openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Decoding protein networks during virus entry by quantitative proteomics.
Article2018-06-13T05:26:34ZVirus entry into host cells relies on interactions between viral and host structures including lipids, carbohydrates and proteins. Particularly, protein-protein interactions between viral surface proteins and host proteins as well as secondary host protein-protein interactions play a pivotal role in coordinating virus binding and uptake. These interactions are dynamic and frequently involve multiprotein complexes. In the past decade mass spectrometry based proteomics methods have reached sensitivities and high throughput compatibilities of genomics methods and now allow the reliable quantitation of proteins in complex samples from limited material. As proteomics provides essential information on the biologically active entity namely the protein, including its posttranslational modifications and its interactions with other proteins, it is an indispensable method in the virologist's toolbox. Here we review protein interactions during virus entry and compare classical biochemical methods to study entry with novel technically advanced quantitative proteomics techniques. We highlight the value of quantitative proteomics in mapping functional virus entry networks, discuss the benefits and limitations and illustrate how the methodology will help resolve unsettled questions in virus entry research in the future.oai:repository.helmholtz-hzi.de:10033/6169242019-08-30T11:35:13Zcom_10033_620589col_10033_620590
Pfaender, Stephanie
Grabski, Elena
Detje, Claudia N
Riebesehl, Nina
Lienenklaus, Stefan
Steinmann, E
Kalinke, Ulrich
Pietschmann, T
Twincore Centre of Experimental and Clinical Infection Research; a joint venture between the Hannover Medical School and the Helmholtz Centre for Infection Research, Hannover 30625, Germany.
2016-07-14T08:05:16Z
2016-07-14T08:05:16Z
2016-07
Hepatitis C Virus Stimulates Murine CD8α-Like Dendritic Cells to Produce Type I Interferon in a TRIF-Dependent Manner. 2016, 12 (7):e1005736 PLoS Pathog.
1553-7374
27385030
10.1371/journal.ppat.1005736
http://hdl.handle.net/10033/616924
PLoS pathogens
Hepatitis C virus (HCV) induces interferon (IFN) stimulated genes in the liver despite of distinct innate immune evasion mechanisms, suggesting that beyond HCV infected cells other cell types contribute to innate immune activation. Upon coculture with HCV replicating cells, human CD141+ myeloid dendritic cells (DC) produce type III IFN, whereas plasmacytoid dendritic cells (pDC) mount type I IFN responses. Due to limitations in the genetic manipulation of primary human DCs, we explored HCV mediated stimulation of murine DC subsets. Coculture of HCV RNA transfected human or murine hepatoma cells with murine bone marrow-derived DC cultures revealed that only Flt3-L DC cultures, but not GM-CSF DC cultures responded with IFN production. Cells transfected with full length or subgenomic viral RNA stimulated IFN release indicating that infectious virus particle formation is not essential in this process. Use of differentiated DC from mice with genetic lesions in innate immune signalling showed that IFN secretion by HCV-stimulated murine DC was independent of MyD88 and CARDIF, but dependent on TRIF and IFNAR signalling. Separating Flt3-L DC cultures into pDC and conventional CD11b-like and CD8α-like DC revealed that the CD8α-like DC, homologous to the human CD141+ DC, release interferon upon stimulation by HCV replicating cells. In contrast, the other cell types and in particular the pDC did not. Injection of human HCV subgenomic replicon cells into IFN-β reporter mice confirmed the interferon induction upon HCV replication in vivo. These results indicate that HCV-replicating cells stimulate IFN secretion from murine CD8α-like DC independent of infectious virus production. Thus, this work defines basic principles of viral recognition by murine DC populations. Moreover, this model should be useful to explore the interaction between dendritic cells during HCV replication and to define how viral signatures are delivered to and recognized by immune cells to trigger IFN release.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Hepatitis C Virus Stimulates Murine CD8α-Like Dendritic Cells to Produce Type I Interferon in a TRIF-Dependent Manner.
Article2018-06-13T03:58:19ZHepatitis C virus (HCV) induces interferon (IFN) stimulated genes in the liver despite of distinct innate immune evasion mechanisms, suggesting that beyond HCV infected cells other cell types contribute to innate immune activation. Upon coculture with HCV replicating cells, human CD141+ myeloid dendritic cells (DC) produce type III IFN, whereas plasmacytoid dendritic cells (pDC) mount type I IFN responses. Due to limitations in the genetic manipulation of primary human DCs, we explored HCV mediated stimulation of murine DC subsets. Coculture of HCV RNA transfected human or murine hepatoma cells with murine bone marrow-derived DC cultures revealed that only Flt3-L DC cultures, but not GM-CSF DC cultures responded with IFN production. Cells transfected with full length or subgenomic viral RNA stimulated IFN release indicating that infectious virus particle formation is not essential in this process. Use of differentiated DC from mice with genetic lesions in innate immune signalling showed that IFN secretion by HCV-stimulated murine DC was independent of MyD88 and CARDIF, but dependent on TRIF and IFNAR signalling. Separating Flt3-L DC cultures into pDC and conventional CD11b-like and CD8α-like DC revealed that the CD8α-like DC, homologous to the human CD141+ DC, release interferon upon stimulation by HCV replicating cells. In contrast, the other cell types and in particular the pDC did not. Injection of human HCV subgenomic replicon cells into IFN-β reporter mice confirmed the interferon induction upon HCV replication in vivo. These results indicate that HCV-replicating cells stimulate IFN secretion from murine CD8α-like DC independent of infectious virus production. Thus, this work defines basic principles of viral recognition by murine DC populations. Moreover, this model should be useful to explore the interaction between dendritic cells during HCV replication and to define how viral signatures are delivered to and recognized by immune cells to trigger IFN release.oai:repository.helmholtz-hzi.de:10033/6205442019-08-30T11:36:04Zcom_10033_620589col_10033_620590
Anggakusuma
Brown, Richard J P
Banda, Dominic
Todt, Daniel
Vieyres, Gabrielle
Steinmann, Eike
Pietschmann, Thomas
Twincore Centre of Experimental and Clinical Infection Research; a joint venture between the Hannover Medical School and the Helmholtz Centre for Infection Research, Hannover 30625, Germany.
2016-10-07T14:33:24Z
2016-10-07T14:33:24Z
2016-09-21
HepacivirusNS3/4A proteases interfere with MAVS signalling of their cognate animal hosts and also with human MAVS: implications for zoonotic transmission. 2016: J. Virol.
1098-5514
27654291
10.1128/JVI.01634-16
http://hdl.handle.net/10033/620544
Journal of virology
Multiple novel members of the genus Hepacivirus have recently been discovered in diverse mammalian species. However, to date, their replication mechanisms and zoonotic potential have not been explored in detail. The NS3/4A serine protease of HCV is critical for cleavage of the viral polyprotein. It also cleaves the cellular innate immune adaptor MAVS, thus decreasing IFN production and contributing to HCV persistence in the human host.To investigate conservation of fundamental aspects of the hepaciviral life-cycle, we explored if MAVS cleavage and suppression of innate immune signaling represents a common mechanism employed across different clades of the genus Hepacivirus to enhance viral replication. To estimate the zoonotic potential of these non-human hepaciviruses, we assessed if their NS3/4A proteases were capable of cleaving human MAVS.NS3/4A proteases of viruses infecting Colobus monkeys, rodents, horses, and cows cleaved the MAVS protein of their cognate hosts and interfered with its ability to induce the IFN-β promoter. All NS3/4A proteases from non-human viruses readily cleaved human MAVS. Thus, NS3/4A-dependent cleavage of MAVS is a conserved replication strategy across multiple clades within the genus Hepacivirus Human MAVS is susceptible to cleavage by these non-human viral proteases indicating that it does not pose a barrier for zoonotic transmission of these viruses to humans.
ENG
http://creativecommons.org/licenses/by-nc-sa/4.0/
Hepacivirus NS3/4A proteases interfere with MAVS signalling of their cognate animal hosts and also with human MAVS: implications for zoonotic transmission.
Article2017-03-21T00:00:00ZMultiple novel members of the genus Hepacivirus have recently been discovered in diverse mammalian species. However, to date, their replication mechanisms and zoonotic potential have not been explored in detail. The NS3/4A serine protease of HCV is critical for cleavage of the viral polyprotein. It also cleaves the cellular innate immune adaptor MAVS, thus decreasing IFN production and contributing to HCV persistence in the human host.To investigate conservation of fundamental aspects of the hepaciviral life-cycle, we explored if MAVS cleavage and suppression of innate immune signaling represents a common mechanism employed across different clades of the genus Hepacivirus to enhance viral replication. To estimate the zoonotic potential of these non-human hepaciviruses, we assessed if their NS3/4A proteases were capable of cleaving human MAVS.NS3/4A proteases of viruses infecting Colobus monkeys, rodents, horses, and cows cleaved the MAVS protein of their cognate hosts and interfered with its ability to induce the IFN-β promoter. All NS3/4A proteases from non-human viruses readily cleaved human MAVS. Thus, NS3/4A-dependent cleavage of MAVS is a conserved replication strategy across multiple clades within the genus Hepacivirus Human MAVS is susceptible to cleavage by these non-human viral proteases indicating that it does not pose a barrier for zoonotic transmission of these viruses to humans.oai:repository.helmholtz-hzi.de:10033/6205492019-08-30T11:35:39Zcom_10033_620589col_10033_620590
Lüdtke, Anja
Ruibal, Paula
Becker-Ziaja, Beate
Rottstegge, Monika
Wozniak, David M
Cabeza-Cabrerizo, Mar
Thorenz, Anja
Weller, Romy
Kerber, Romy
Idoyaga, Juliana
Magassouba, N'Faly
Gabriel, Martin
Günther, Stephan
Oestereich, Lisa
Muñoz-Fontela, César
TwinCore, Centre for experimental and clinical infection research GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2016-10-12T13:56:47Z
2016-10-12T13:56:47Z
2016-10-15
Ebola Virus Disease Is Characterized by Poor Activation and Reduced Levels of Circulating CD16+ Monocytes. 2016, 214 (suppl 3):S275-S280 J. Infect. Dis.
1537-6613
27521367
10.1093/infdis/jiw260
http://hdl.handle.net/10033/620549
The Journal of infectious diseases
A number of previous studies have identified antigen-presenting cells (APCs) as key targets of Ebola virus (EBOV), but the role of APCs in human Ebola virus disease (EVD) is not known. We have evaluated the phenotype and kinetics of monocytes, neutrophils, and dendritic cells (DCs) in peripheral blood of patients for whom EVD was diagnosed by the European Mobile Laboratory in Guinea. Acute EVD was characterized by reduced levels of circulating nonclassical CD16(+) monocytes with a poor activation profile. In survivors, CD16(+) monocytes were activated during recovery, coincident with viral clearance, suggesting an important role of this cell subset in EVD pathophysiology.
ENG
'info:eu-repo/grantAgreement/EC/FP7/'.Example: info:eu-repo/grantAgreement/EC/H2020/666100
openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Ebola Virus Disease Is Characterized by Poor Activation and Reduced Levels of Circulating CD16+ Monocytes.
Article2018-06-13T09:15:21ZA number of previous studies have identified antigen-presenting cells (APCs) as key targets of Ebola virus (EBOV), but the role of APCs in human Ebola virus disease (EVD) is not known. We have evaluated the phenotype and kinetics of monocytes, neutrophils, and dendritic cells (DCs) in peripheral blood of patients for whom EVD was diagnosed by the European Mobile Laboratory in Guinea. Acute EVD was characterized by reduced levels of circulating nonclassical CD16(+) monocytes with a poor activation profile. In survivors, CD16(+) monocytes were activated during recovery, coincident with viral clearance, suggesting an important role of this cell subset in EVD pathophysiology.oai:repository.helmholtz-hzi.de:10033/6205532019-08-30T11:33:53Zcom_10033_620589col_10033_620590
Bankwitz, Dorothea
Pietschmann, Thomas
TwinCore, Centre for experimental and clinical infection research GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2016-10-17T13:17:13Z
2016-10-17T13:17:13Z
2016-08-12
Hepatitis C virus plays hide and seek with neutralizing antibodies. 2016 Hepatology
1527-3350
27515101
10.1002/hep.28760
http://hdl.handle.net/10033/620553
Hepatology (Baltimore, Md.)
ENG
http://creativecommons.org/licenses/by-nc-sa/4.0/
Hepatitis C virus plays hide and seek with neutralizing antibodies.
Article2018-06-12T17:58:56Zoai:repository.helmholtz-hzi.de:10033/6205832019-08-30T11:26:13Zcom_10033_620589col_10033_620590
Ruibal, Paula
Oestereich, Lisa
Lüdtke, Anja
Becker-Ziaja, Beate
Wozniak, David M
Kerber, Romy
Korva, Miša
Cabeza-Cabrerizo, Mar
Bore, Joseph A
Koundouno, Fara Raymond
Duraffour, Sophie
Weller, Romy
Thorenz, Anja
Cimini, Eleonora
Viola, Domenico
Agrati, Chiara
Repits, Johanna
Afrough, Babak
Cowley, Lauren A
Ngabo, Didier
Hinzmann, Julia
Mertens, Marc
Vitoriano, Inês
Logue, Christopher H
Boettcher, Jan Peter
Pallasch, Elisa
Sachse, Andreas
Bah, Amadou
Nitzsche, Katja
Kuisma, Eeva
Michel, Janine
Holm, Tobias
Zekeng, Elsa-Gayle
García-Dorival, Isabel
Wölfel, Roman
Stoecker, Kilian
Fleischmann, Erna
Strecker, Thomas
Di Caro, Antonino
Avšič-Županc, Tatjana
Kurth, Andreas
Meschi, Silvia
Mély, Stephane
Newman, Edmund
Bocquin, Anne
Kis, Zoltan
Kelterbaum, Anne
Molkenthin, Peter
Carletti, Fabrizio
Portmann, Jasmine
Wolff, Svenja
Castilletti, Concetta
Schudt, Gordian
Fizet, Alexandra
Ottowell, Lisa J
Herker, Eva
Jacobs, Thomas
Kretschmer, Birte
Severi, Ettore
Ouedraogo, Nobila
Lago, Mar
Negredo, Anabel
Franco, Leticia
Anda, Pedro
Schmiedel, Stefan
Kreuels, Benno
Wichmann, Dominic
Addo, Marylyn M
Lohse, Ansgar W
De Clerck, Hilde
Nanclares, Carolina
Jonckheere, Sylvie
Van Herp, Michel
Sprecher, Armand
Xiaojiang, Gao
Carrington, Mary
Miranda, Osvaldo
Castro, Carlos M
Gabriel, Martin
Drury, Patrick
Formenty, Pierre
Diallo, Boubacar
Koivogui, Lamine
Magassouba, N'Faly
Carroll, Miles W
Günther, Stephan
Muñoz-Fontela, César
TWINCORE, Centre for experimental and clinical infection research GmbH, Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2016-11-17T12:19:23Z
2016-11-17T12:19:23Z
2016-05-05
Unique human immune signature of Ebola virus disease in Guinea. 2016, 533 (7601):100-4 Nature
0028-0836
27147028
10.1038/nature17949
http://hdl.handle.net/10033/620583
Nature
Despite the magnitude of the Ebola virus disease (EVD) outbreak in West Africa, there is still a fundamental lack of knowledge about the pathophysiology of EVD. In particular, very little is known about human immune responses to Ebola virus. Here we evaluate the physiology of the human T cell immune response in EVD patients at the time of admission to the Ebola Treatment Center in Guinea, and longitudinally until discharge or death. Through the use of multiparametric flow cytometry established by the European Mobile Laboratory in the field, we identify an immune signature that is unique in EVD fatalities. Fatal EVD was characterized by a high percentage of CD4(+) and CD8(+) T cells expressing the inhibitory molecules CTLA-4 and PD-1, which correlated with elevated inflammatory markers and high virus load. Conversely, surviving individuals showed significantly lower expression of CTLA-4 and PD-1 as well as lower inflammation, despite comparable overall T cell activation. Concomitant with virus clearance, survivors mounted a robust Ebola-virus-specific T cell response. Our findings suggest that dysregulation of the T cell response is a key component of EVD pathophysiology.
ENG
: info:eu-repo/grantAgreement/H2020/666100
openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
CTLA-4 Antigen
Ebolavirus
Female
Flow Cytometry
Guinea
Hemorrhagic Fever, Ebola
Humans
Inflammation Mediators
Longitudinal Studies
Lymphocyte Activation
Male
Patient Discharge
Programmed Cell Death 1 Receptor
Survivors
T-Lymphocytes
Viral Load
Unique human immune signature of Ebola virus disease in Guinea.
Article2018-06-13T02:43:04ZDespite the magnitude of the Ebola virus disease (EVD) outbreak in West Africa, there is still a fundamental lack of knowledge about the pathophysiology of EVD. In particular, very little is known about human immune responses to Ebola virus. Here we evaluate the physiology of the human T cell immune response in EVD patients at the time of admission to the Ebola Treatment Center in Guinea, and longitudinally until discharge or death. Through the use of multiparametric flow cytometry established by the European Mobile Laboratory in the field, we identify an immune signature that is unique in EVD fatalities. Fatal EVD was characterized by a high percentage of CD4(+) and CD8(+) T cells expressing the inhibitory molecules CTLA-4 and PD-1, which correlated with elevated inflammatory markers and high virus load. Conversely, surviving individuals showed significantly lower expression of CTLA-4 and PD-1 as well as lower inflammation, despite comparable overall T cell activation. Concomitant with virus clearance, survivors mounted a robust Ebola-virus-specific T cell response. Our findings suggest that dysregulation of the T cell response is a key component of EVD pathophysiology.oai:repository.helmholtz-hzi.de:10033/6209012019-08-30T11:33:29Zcom_10033_620589col_10033_620590
Pietschmann, Thomas
TwinCore, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2017-04-21T10:35:55Z
2017-04-21T10:35:55Z
2017-01-15
Clinically Approved Ion Channel Inhibitors Close Gates for Hepatitis C Virus and Open Doors for Drug Repurposing in Infectious Viral Diseases. 2017, 91 (2) J. Virol.
1098-5514
27807238
10.1128/JVI.01914-16
http://hdl.handle.net/10033/620901
Journal of virology
Chronic hepatitis C virus (HCV) infection causes severe liver disease and affects ca. 146 million individuals. Novel directly acting antivirals targeting HCV have revolutionized treatment. However, high costs limit access to therapy. Recently, several related drugs used in humans to treat allergies or as neuroleptics emerged as potent HCV cell entry inhibitors. Insights into their antiviral modes of action may increase opportunities for drug repurposing in hepatitis C and possibly other important human viral infections.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Clinically Approved Ion Channel Inhibitors Close Gates for Hepatitis C Virus and Open Doors for Drug Repurposing in Infectious Viral Diseases.
Article2017-05-02T00:00:00ZChronic hepatitis C virus (HCV) infection causes severe liver disease and affects ca. 146 million individuals. Novel directly acting antivirals targeting HCV have revolutionized treatment. However, high costs limit access to therapy. Recently, several related drugs used in humans to treat allergies or as neuroleptics emerged as potent HCV cell entry inhibitors. Insights into their antiviral modes of action may increase opportunities for drug repurposing in hepatitis C and possibly other important human viral infections.oai:repository.helmholtz-hzi.de:10033/6209362019-08-30T11:33:57Zcom_10033_620589col_10033_620590
Cimini, Eleonora
Viola, Domenico
Cabeza-Cabrerizo, Mar
Romanelli, Antonella
Tumino, Nicola
Sacchi, Alessandra
Bordoni, Veronica
Casetti, Rita
Turchi, Federica
Martini, Federico
Bore, Joseph A
Koundouno, Fara Raymond
Duraffour, Sophie
Michel, Janine
Holm, Tobias
Zekeng, Elsa Gayle
Cowley, Lauren
Garcia Dorival, Isabel
Doerrbecker, Juliane
Hetzelt, Nicole
Baum, Jonathan H J
Portmann, Jasmine
Wölfel, Roman
Gabriel, Martin
Miranda, Osvaldo
Díaz, Graciliano
Díaz, José E
Fleites, Yoel A
Piñeiro, Carlos A
Castro, Carlos M
Koivogui, Lamine
Magassouba, N'Faly
Diallo, Boubacar
Ruibal, Paula
Oestereich, Lisa
Wozniak, David M
Lüdtke, Anja
Becker-Ziaja, Beate
Capobianchi, Maria R
Ippolito, Giuseppe
Carroll, Miles W
Günther, Stephan
Di Caro, Antonino
Muñoz-Fontela, César
Agrati, Chiara
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung gmbH, Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2017-06-09T08:50:46Z
2017-06-09T08:50:46Z
2017-05-30
Different features of Vδ2 T and NK cells in fatal and non-fatal human Ebola infections. 2017, 11 (5):e0005645 PLoS Negl Trop Dis
1935-2735
28558022
10.1371/journal.pntd.0005645
http://hdl.handle.net/10033/620936
PLoS neglected tropical diseases
Human Ebola infection is characterized by a paralysis of the immune system. A signature of αβ T cells in fatal Ebola infection has been recently proposed, while the involvement of innate immune cells in the protection/pathogenesis of Ebola infection is unknown. Aim of this study was to analyze γδ T and NK cells in patients from the Ebola outbreak of 2014-2015 occurred in West Africa, and to assess their association with the clinical outcome.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Different features of Vδ2 T and NK cells in fatal and non-fatal human Ebola infections.
Article2018-06-13T05:36:19ZHuman Ebola infection is characterized by a paralysis of the immune system. A signature of αβ T cells in fatal Ebola infection has been recently proposed, while the involvement of innate immune cells in the protection/pathogenesis of Ebola infection is unknown. Aim of this study was to analyze γδ T and NK cells in patients from the Ebola outbreak of 2014-2015 occurred in West Africa, and to assess their association with the clinical outcome.oai:repository.helmholtz-hzi.de:10033/6209732019-08-30T11:34:48Zcom_10033_620589col_10033_620590col_10033_620596
Siddharta, Anindya
Pfaender, Stephanie
Vielle, Nathalie Jane
Dijkman, Ronald
Friesland, Martina
Becker, Britta
Yang, Jaewon
Engelmann, Michael
Todt, Daniel
Windisch, Marc P
Brill, Florian H
Steinmann, Joerg
Steinmann, Jochen
Becker, Stephan
Alves, Marco P
Pietschmann, Thomas
Eickmann, Markus
Thiel, Volker
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung gmbH, Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2017-06-22T09:40:09Z
2017-06-22T09:40:09Z
2017-03-15
Virucidal Activity of World Health Organization-Recommended Formulations Against Enveloped Viruses, Including Zika, Ebola, and Emerging Coronaviruses. 2017, 215 (6):902-906 J. Infect. Dis.
1537-6613
28453839
10.1093/infdis/jix046
http://hdl.handle.net/10033/620973
The Journal of infectious diseases
The World Health Organization (WHO) published 2 alcohol-based formulations to be used in healthcare settings and for outbreak-associated infections, but inactivation efficacies of these products have not been determined against (re-)emerging viruses. In this study, we evaluated the virucidal activity of these WHO products in a comparative analysis. Zika virus (ZIKV), Ebola virus (EBOV), severe acute respiratory syndrome coronavirus (SARS-CoV), and Middle East respiratory syndrome coronavirus (MERS-CoV) as (re-)emerging viral pathogens and other enveloped viruses could be efficiently inactivated by both WHO formulations, implicating their use in healthcare systems and viral outbreak situations.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Virucidal Activity of World Health Organization-Recommended Formulations Against Enveloped Viruses, Including Zika, Ebola, and Emerging Coronaviruses.
Article2018-06-12T16:52:37ZThe World Health Organization (WHO) published 2 alcohol-based formulations to be used in healthcare settings and for outbreak-associated infections, but inactivation efficacies of these products have not been determined against (re-)emerging viruses. In this study, we evaluated the virucidal activity of these WHO products in a comparative analysis. Zika virus (ZIKV), Ebola virus (EBOV), severe acute respiratory syndrome coronavirus (SARS-CoV), and Middle East respiratory syndrome coronavirus (MERS-CoV) as (re-)emerging viral pathogens and other enveloped viruses could be efficiently inactivated by both WHO formulations, implicating their use in healthcare systems and viral outbreak situations.oai:repository.helmholtz-hzi.de:10033/6209812019-08-30T11:34:48Zcom_10033_620589col_10033_620590
Bankwitz, Dorothea
Doepke, Mandy
Hueging, Kathrin
Weller, Romy
Bruening, Janina
Behrendt, Patrick
Lee, Ji-Young
Vondran, Florian W R
Manns, Michael P
Bartenschlager, Ralf
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen Str.7, 30625 Hannover, Germany.
2017-06-26T14:37:58Z
2017-06-26T14:37:58Z
2017-04-22
Maturation of secreted HCV particles by incorporation of secreted ApoE protects from antibodies by enhancing infectivity. 2017 J. Hepatol.
1600-0641
28438690
10.1016/j.jhep.2017.04.010
http://hdl.handle.net/10033/620981
Journal of hepatology
Hepatitis C virus (HCV) evades humoral immunity and establishes chronic infections. Virus particles circulate in complex with lipoproteins facilitating antibody escape. Apolipoprotein E (ApoE) is essential for intracellular HCV assembly and for HCV cell entry. We aimed to explore if ApoE released from non-infected cells interacts with and modulates secreted HCV particles.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Maturation of secreted HCV particles by incorporation of secreted ApoE protects from antibodies by enhancing infectivity.
Article2018-06-14T09:19:28ZHepatitis C virus (HCV) evades humoral immunity and establishes chronic infections. Virus particles circulate in complex with lipoproteins facilitating antibody escape. Apolipoprotein E (ApoE) is essential for intracellular HCV assembly and for HCV cell entry. We aimed to explore if ApoE released from non-infected cells interacts with and modulates secreted HCV particles.oai:repository.helmholtz-hzi.de:10033/6210092019-08-30T11:28:51Zcom_10033_620589com_10033_620636col_10033_620590col_10033_620596col_10033_620638
Khera, Tanvi
Todt, Daniel
Vercauteren, Koen
McClure, C Patrick
Verhoye, Lieven
Farhoudi, Ali
Bhuju, Sabin
Geffers, Robert
Baumert, Thomas F
Steinmann, Eike
Meuleman, Philip
Pietschmann, Thomas
Brown, Richard J P
Helmholtz Centre for infection research, Inhoffenstr. 7., 38124 Braunschweig, Germany.
2017-07-12T14:20:02Z
2017-07-12T14:20:02Z
2017-03
Tracking HCV protease population diversity during transmission and susceptibility of founder populations to antiviral therapy. 2017, 139:129-137 Antiviral Res.
1872-9096
28062191
10.1016/j.antiviral.2017.01.001
http://hdl.handle.net/10033/621009
Antiviral research
Due to the highly restricted species-tropism of Hepatitis C virus (HCV) a limited number of animal models exist for pre-clinical evaluation of vaccines and antiviral compounds. The human-liver chimeric mouse model allows heterologous challenge with clinically relevant strains derived from patients. However, to date, the transmission and longitudinal evolution of founder viral populations in this model have not been characterized in-depth using state-of-the-art sequencing technologies. Focusing on NS3 protease encoding region of the viral genome, mutant spectra in a donor inoculum and individual recipient mice were determined via Illumina sequencing and compared, to determine the effects of transmission on founder viral population complexity. In all transmissions, a genetic bottleneck was observed, although diverse viral populations were transmitted in each case. A low frequency cloud of mutations (<1%) was detectable in the donor inoculum and recipient mice, with single nucleotide variants (SNVs) > 1% restricted to a subset of nucleotides. The population of SNVs >1% was reduced upon transmission while the low frequency SNV cloud remained stable. Fixation of multiple identical synonymous substitutions was apparent in independent transmissions, and no evidence for reversion of T-cell epitopes was observed. In addition, susceptibility of founder populations to antiviral therapy was assessed. Animals were treated with protease inhibitor (PI) monotherapy to track resistance associated substitution (RAS) emergence. Longitudinal analyses revealed a decline in population diversity under therapy, with no detectable RAS >1% prior to therapy commencement. Despite inoculation from a common source and identical therapeutic regimens, unique RAS emergence profiles were identified in different hosts prior to and during therapeutic failure, with complex mutational signatures at protease residues 155, 156 and 168 detected. Together these analyses track viral population complexity at high-resolution in the human-liver chimeric mouse model post-transmission and under therapeutic intervention, revealing novel insights into the evolutionary processes which shape viral protease population composition at various critical stages of the viral life-cycle.
en
info:eu-repo/grantAgreement/EC/FP7/305600
openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Tracking HCV protease population diversity during transmission and susceptibility of founder populations to antiviral therapy.
Article2018-01-03T00:00:00ZDue to the highly restricted species-tropism of Hepatitis C virus (HCV) a limited number of animal models exist for pre-clinical evaluation of vaccines and antiviral compounds. The human-liver chimeric mouse model allows heterologous challenge with clinically relevant strains derived from patients. However, to date, the transmission and longitudinal evolution of founder viral populations in this model have not been characterized in-depth using state-of-the-art sequencing technologies. Focusing on NS3 protease encoding region of the viral genome, mutant spectra in a donor inoculum and individual recipient mice were determined via Illumina sequencing and compared, to determine the effects of transmission on founder viral population complexity. In all transmissions, a genetic bottleneck was observed, although diverse viral populations were transmitted in each case. A low frequency cloud of mutations (<1%) was detectable in the donor inoculum and recipient mice, with single nucleotide variants (SNVs) > 1% restricted to a subset of nucleotides. The population of SNVs >1% was reduced upon transmission while the low frequency SNV cloud remained stable. Fixation of multiple identical synonymous substitutions was apparent in independent transmissions, and no evidence for reversion of T-cell epitopes was observed. In addition, susceptibility of founder populations to antiviral therapy was assessed. Animals were treated with protease inhibitor (PI) monotherapy to track resistance associated substitution (RAS) emergence. Longitudinal analyses revealed a decline in population diversity under therapy, with no detectable RAS >1% prior to therapy commencement. Despite inoculation from a common source and identical therapeutic regimens, unique RAS emergence profiles were identified in different hosts prior to and during therapeutic failure, with complex mutational signatures at protease residues 155, 156 and 168 detected. Together these analyses track viral population complexity at high-resolution in the human-liver chimeric mouse model post-transmission and under therapeutic intervention, revealing novel insights into the evolutionary processes which shape viral protease population composition at various critical stages of the viral life-cycle.oai:repository.helmholtz-hzi.de:10033/6210102019-08-30T11:36:32Zcom_10033_620601com_10033_620589col_10033_620602col_10033_620590col_10033_620590
Pfaender, Stephanie
Walter, Stephanie
Grabski, Elena
Todt, Daniel
Bruening, Janina
Romero-Brey, Inés
Gather, Theresa
Brown, Richard J P
Hahn, Kerstin
Puff, Christina
Pfankuche, Vanessa M
Hansmann, Florian
Postel, Alexander
Becher, Paul
Thiel, Volker
Kalinke, Ulrich
Wagner, Bettina
Bartenschlager, Ralf
Baumgärtner, Wolfgang
Feige, Karsten
Pietschmann, Thomas
Cavalleri, Jessika M V
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infectionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2017-07-13T11:52:32Z
2017-07-13T11:52:32Z
2017-03-21
Immune protection against reinfection with nonprimate hepacivirus. 2017, 114 (12):E2430-E2439 Proc. Natl. Acad. Sci. U.S.A.
1091-6490
28275093
10.1073/pnas.1619380114
http://hdl.handle.net/10033/621010
Proceedings of the National Academy of Sciences of the United States of America
Hepatitis C virus (HCV) displays a restricted host species tropism and only humans and chimpanzees are susceptible to infection. A robust immunocompetent animal model is still lacking, hampering mechanistic analysis of virus pathogenesis, immune control, and prophylactic vaccine development. The closest homolog of HCV is the equine nonprimate hepacivirus (NPHV), which shares similar features with HCV and thus represents an animal model to study hepacivirus infections in their natural hosts. We aimed to dissect equine immune responses after experimental NPHV infection and conducted challenge experiments to investigate immune protection against secondary NPHV infections. Horses were i.v. injected with NPHV containing plasma. Flow cytometric analysis was used to monitor immune cell frequencies and activation status. All infected horses became viremic after 1 or 2 wk and viremia could be detected in two horses for several weeks followed by a delayed seroconversion and viral clearance. Histopathological examinations of liver biopsies revealed mild, periportally accentuated infiltrations of lymphocytes, macrophages, and plasma cells with some horses displaying subclinical signs of hepatitis. Following viral challenge, an activation of equine immune responses was observed. Importantly, after a primary NPHV infection, horses were protected against rechallenge with the homologous as well as a distinct isolate with only minute amounts of circulating virus being detectable.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Immune protection against reinfection with nonprimate hepacivirus.
Article2018-06-13T04:23:48ZHepatitis C virus (HCV) displays a restricted host species tropism and only humans and chimpanzees are susceptible to infection. A robust immunocompetent animal model is still lacking, hampering mechanistic analysis of virus pathogenesis, immune control, and prophylactic vaccine development. The closest homolog of HCV is the equine nonprimate hepacivirus (NPHV), which shares similar features with HCV and thus represents an animal model to study hepacivirus infections in their natural hosts. We aimed to dissect equine immune responses after experimental NPHV infection and conducted challenge experiments to investigate immune protection against secondary NPHV infections. Horses were i.v. injected with NPHV containing plasma. Flow cytometric analysis was used to monitor immune cell frequencies and activation status. All infected horses became viremic after 1 or 2 wk and viremia could be detected in two horses for several weeks followed by a delayed seroconversion and viral clearance. Histopathological examinations of liver biopsies revealed mild, periportally accentuated infiltrations of lymphocytes, macrophages, and plasma cells with some horses displaying subclinical signs of hepatitis. Following viral challenge, an activation of equine immune responses was observed. Importantly, after a primary NPHV infection, horses were protected against rechallenge with the homologous as well as a distinct isolate with only minute amounts of circulating virus being detectable.oai:repository.helmholtz-hzi.de:10033/6210772019-08-30T11:36:33Zcom_10033_620589com_10033_311308col_10033_620721col_10033_620590col_10033_620596
Pfaender, Stephanie
Walter, Stephanie
Todt, Daniel
Behrendt, Patrick
Doerrbecker, Juliane
Wölk, Benno
Engelmann, Michael
Gravemann, Ute
Seltsam, Axel
Steinmann, Joerg
Burbelo, Peter D
Klawonn, Frank
Feige, Karsten
Pietschmann, Thomas
Cavalleri, Jessika-M V
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2017-08-29T08:45:06Z
2017-08-29T08:45:06Z
2015-09
Assessment of cross-species transmission of hepatitis C virus-related non-primate hepacivirus in a population of humans at high risk of exposure. 2015, 96 (9):2636-42 J. Gen. Virol.
1465-2099
26041875
10.1099/vir.0.000208
http://hdl.handle.net/10033/621077
The Journal of general virology
The recent discovery of hepatitis C virus (HCV)-related viruses in different animal species has raised new speculations regarding the origin of HCV and the possibility of a zoonotic source responsible for the endemic HCV transmission. As a consequence, these new findings prompt questions regarding the potential for cross-species transmissions of hepaciviruses. The closest relatives to HCV discovered to date are the non-primate hepaciviruses (NPHVs), which have been described to infect horses. To evaluate the risk of a potential zoonotic transmission, we analysed NPHV RNA and antibodies in humans with occupational exposure to horses in comparison with a low-risk group. Both groups were negative for NPHV RNA, even though low seroreactivities against various NPHV antigens could be detected irrespective of the group. In conclusion, we did not observe evidence of NPHV transmission between horses and humans.
en
info:eu-repo/grantAgreement/EC/FP7/281473
openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Adult
Agricultural Workers' Diseases
Animals
Female
Hepacivirus
Hepatitis C
Horse Diseases
Horses
Humans
Male
Middle Aged
Occupational Exposure
Phylogeny
Zoonoses
Assessment of cross-species transmission of hepatitis C virus-related non-primate hepacivirus in a population of humans at high risk of exposure.
Article2018-06-12T22:01:42ZThe recent discovery of hepatitis C virus (HCV)-related viruses in different animal species has raised new speculations regarding the origin of HCV and the possibility of a zoonotic source responsible for the endemic HCV transmission. As a consequence, these new findings prompt questions regarding the potential for cross-species transmissions of hepaciviruses. The closest relatives to HCV discovered to date are the non-primate hepaciviruses (NPHVs), which have been described to infect horses. To evaluate the risk of a potential zoonotic transmission, we analysed NPHV RNA and antibodies in humans with occupational exposure to horses in comparison with a low-risk group. Both groups were negative for NPHV RNA, even though low seroreactivities against various NPHV antigens could be detected irrespective of the group. In conclusion, we did not observe evidence of NPHV transmission between horses and humans.oai:repository.helmholtz-hzi.de:10033/6212292019-08-30T11:33:57Zcom_10033_620589col_10033_620590
Bruening, Janina
Weigel, Bettina
Gerold, Gisa
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2018-01-09T15:25:20Z
2018-01-09T15:25:20Z
2017
The Role of Type III Interferons in Hepatitis C Virus Infection and Therapy. 2017, 2017:7232361 J Immunol Res
2314-7156
28255563
10.1155/2017/7232361
http://hdl.handle.net/10033/621229
Journal of immunology research
The human interferon (IFN) response is a key innate immune mechanism to fight virus infection. IFNs are host-encoded secreted proteins, which induce IFN-stimulated genes (ISGs) with antiviral properties. Among the three classes of IFNs, type III IFNs, also called IFN lambdas (IFNLs), are an essential component of the innate immune response to hepatitis C virus (HCV). In particular, human polymorphisms in IFNL gene loci correlate with hepatitis C disease progression and with treatment response. To date, the underlying mechanisms remain mostly elusive; however it seems clear that viral infection of the liver induces IFNL responses. As IFNL receptors show a more restricted tissue expression than receptors for other classes of IFNs, IFNL treatment has reduced side effects compared to the classical type I IFN treatment. In HCV therapy, however, IFNL will likely not play an important role as highly effective direct acting antivirals (DAA) exist. Here, we will review our current knowledge on IFNL gene expression, protein properties, signaling, ISG induction, and its implications on HCV infection and treatment. Finally, we will discuss the lessons learnt from the HCV and IFNL field for virus infections beyond hepatitis C.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Animals
Antiviral Agents
Hepacivirus
Hepatitis C
Host-Pathogen Interactions
Humans
Immunity, Innate
Interferons
Interleukins
Liver
Signal Transduction
Virus Replication
The Role of Type III Interferons in Hepatitis C Virus Infection and Therapy.
Article2018-06-12T16:43:11ZThe human interferon (IFN) response is a key innate immune mechanism to fight virus infection. IFNs are host-encoded secreted proteins, which induce IFN-stimulated genes (ISGs) with antiviral properties. Among the three classes of IFNs, type III IFNs, also called IFN lambdas (IFNLs), are an essential component of the innate immune response to hepatitis C virus (HCV). In particular, human polymorphisms in IFNL gene loci correlate with hepatitis C disease progression and with treatment response. To date, the underlying mechanisms remain mostly elusive; however it seems clear that viral infection of the liver induces IFNL responses. As IFNL receptors show a more restricted tissue expression than receptors for other classes of IFNs, IFNL treatment has reduced side effects compared to the classical type I IFN treatment. In HCV therapy, however, IFNL will likely not play an important role as highly effective direct acting antivirals (DAA) exist. Here, we will review our current knowledge on IFNL gene expression, protein properties, signaling, ISG induction, and its implications on HCV infection and treatment. Finally, we will discuss the lessons learnt from the HCV and IFNL field for virus infections beyond hepatitis C.oai:repository.helmholtz-hzi.de:10033/6212572019-08-30T11:28:23Zcom_10033_620589col_10033_620590
Gerold, Gisa
Bruening, Janina
Weigel, Bettina
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2018-01-29T13:53:38Z
2018-01-29T13:53:38Z
2017-04
Protein Interactions during the Flavivirus and Hepacivirus Life Cycle. 2017, 16 (4 suppl 1):S75-S91 Mol. Cell Proteomics
1535-9484
28077444
10.1074/mcp.R116.065649
http://hdl.handle.net/10033/621257
Molecular & cellular proteomics : MCP
Protein-protein interactions govern biological functions in cells, in the extracellular milieu, and at the border between cells and extracellular space. Viruses are small intracellular parasites and thus rely on protein interactions to produce progeny inside host cells and to spread from cell to cell. Usage of host proteins by viruses can have severe consequences e.g. apoptosis, metabolic disequilibria, or altered cell proliferation and mobility. Understanding protein interactions during virus infection can thus educate us on viral infection and pathogenesis mechanisms. Moreover, it has led to important clinical translations, including the development of new therapeutic and vaccination strategies. Here, we will discuss protein interactions of members of the Flaviviridae family, which are small enveloped RNA viruses. Dengue virus, Zika virus and hepatitis C virus belong to the most prominent human pathogenic Flaviviridae With a genome of roughly ten kilobases encoding only ten viral proteins, Flaviviridae display intricate mechanisms to engage the host cell machinery for their purpose. In this review, we will highlight how dengue virus, hepatitis C virus, Japanese encephalitis virus, tick-borne encephalitis virus, West Nile virus, yellow fever virus, and Zika virus proteins engage host proteins and how this knowledge helps elucidate Flaviviridae infection. We will specifically address the protein composition of the virus particle as well as the protein interactions during virus entry, replication, particle assembly, and release from the host cell. Finally, we will give a perspective on future challenges in Flaviviridae interaction proteomics and why we believe these challenges should be met.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Flavivirus
Flavivirus Infections
Genome, Viral
Hepacivirus
Hepatitis C
Host-Pathogen Interactions
Humans
Protein Interaction Maps
Virus Internalization
Virus Replication
Protein Interactions during the Flavivirus and Hepacivirus Life Cycle.
Article2018-04-04T00:00:00ZProtein-protein interactions govern biological functions in cells, in the extracellular milieu, and at the border between cells and extracellular space. Viruses are small intracellular parasites and thus rely on protein interactions to produce progeny inside host cells and to spread from cell to cell. Usage of host proteins by viruses can have severe consequences e.g. apoptosis, metabolic disequilibria, or altered cell proliferation and mobility. Understanding protein interactions during virus infection can thus educate us on viral infection and pathogenesis mechanisms. Moreover, it has led to important clinical translations, including the development of new therapeutic and vaccination strategies. Here, we will discuss protein interactions of members of the Flaviviridae family, which are small enveloped RNA viruses. Dengue virus, Zika virus and hepatitis C virus belong to the most prominent human pathogenic Flaviviridae With a genome of roughly ten kilobases encoding only ten viral proteins, Flaviviridae display intricate mechanisms to engage the host cell machinery for their purpose. In this review, we will highlight how dengue virus, hepatitis C virus, Japanese encephalitis virus, tick-borne encephalitis virus, West Nile virus, yellow fever virus, and Zika virus proteins engage host proteins and how this knowledge helps elucidate Flaviviridae infection. We will specifically address the protein composition of the virus particle as well as the protein interactions during virus entry, replication, particle assembly, and release from the host cell. Finally, we will give a perspective on future challenges in Flaviviridae interaction proteomics and why we believe these challenges should be met.oai:repository.helmholtz-hzi.de:10033/6212612019-08-30T11:35:13Zcom_10033_620589col_10033_620590
Fedeli, Chiara
Torriani, Giulia
Galan-Navarro, Clara
Moraz, Marie-Laurence
Moreno, Hector
Gerold, Gisa
Kunz, Stefan
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
2018-01-30T15:17:34Z
2018-01-30T15:17:34Z
2017-12-13
Axl can serve as entry factor for Lassa virus depending on the functional glycosylation of dystroglycan. 2017 J. Virol.
1098-5514
29237830
10.1128/JVI.01613-17
http://hdl.handle.net/10033/621261
Journal of virology
Fatal infection with the highly pathogenic Lassa virus (LASV) is characterized by extensive viral dissemination, indicating broad tissue tropism. The major cellular receptor for LASV is the highly conserved extracellular matrix receptor dystroglycan (DG). Binding of LASV depends on DG's tissue-specific post-translational modification with the unusual O-linked polysaccharide matriglycan. Interestingly, functional glycosylation of DG does not always correlate with viral tropism observed in vivo The broadly expressed phosphatidylserine (PS) receptors Axl and Tyro3 were recently identified as alternative LASV receptor candidates. However, their role in LASV entry is not entirely understood. Here we examined LASV receptor candidates in primary human cells and found co-expression of Axl with differentially glycosylated DG. To study LASV receptor use in the context of productive arenavirus infection, we employed recombinant lymphocytic choriomeningitis virus expressing LASV glycoprotein (rLCMV-LASVGP) as validated BSL2 model. We confirm and extend previous work, showing that Axl can contribute to LASV entry in absence of functional DG using "apoptotic mimicry", similar to other enveloped virus. We further show that Axl-dependent LASV entry requires receptor activation and involves a pathway resembling macropinocytosis. Axl-mediated LASV entry is facilitated by heparan sulfate and critically depends on the late endosomal protein LAMP-1 as intracellular entry factor. In endothelial cells expressing low levels of functional DG, both receptors are engaged by the virus and can contribute to productive entry. In sum, we characterize the role of Axl in LASV entry and provide a rationale to target Axl in anti-viral therapy.IMPORTANCEThe highly pathogenic arenavirus Lassa (LASV) represents a serious public health problem in Africa. Although the principal LASV receptor dystroglycan (DG) is ubiquitously expressed, virus binding critically depends on DG's post-translational modification, which does not always correlate with tissue tropism. The broadly expressed phosphatidylserine receptor Axl was recently identified as alternative LASV receptor candidate, but its role in LASV entry is unclear. Here we investigated the exact role of Axl in LASV entry as a function of DG's post-translational modification. We found that in absence of functional DG, Axl can mediate LASV entry via "apoptotic mimicry". Productive entry requires virus-induced receptor activation, involves macropinocytosis, and critically depends on LAMP-1. In endothelial cells that express low levels of glycosylated DG, both receptors can promote LASV entry. In sum, our study defines the roles of Axl in LASV entry and provides a rationale to target Axl in anti-viral therapy.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Axl can serve as entry factor for Lassa virus depending on the functional glycosylation of dystroglycan.
Article2018-05-13T00:00:00ZFatal infection with the highly pathogenic Lassa virus (LASV) is characterized by extensive viral dissemination, indicating broad tissue tropism. The major cellular receptor for LASV is the highly conserved extracellular matrix receptor dystroglycan (DG). Binding of LASV depends on DG's tissue-specific post-translational modification with the unusual O-linked polysaccharide matriglycan. Interestingly, functional glycosylation of DG does not always correlate with viral tropism observed in vivo The broadly expressed phosphatidylserine (PS) receptors Axl and Tyro3 were recently identified as alternative LASV receptor candidates. However, their role in LASV entry is not entirely understood. Here we examined LASV receptor candidates in primary human cells and found co-expression of Axl with differentially glycosylated DG. To study LASV receptor use in the context of productive arenavirus infection, we employed recombinant lymphocytic choriomeningitis virus expressing LASV glycoprotein (rLCMV-LASVGP) as validated BSL2 model. We confirm and extend previous work, showing that Axl can contribute to LASV entry in absence of functional DG using "apoptotic mimicry", similar to other enveloped virus. We further show that Axl-dependent LASV entry requires receptor activation and involves a pathway resembling macropinocytosis. Axl-mediated LASV entry is facilitated by heparan sulfate and critically depends on the late endosomal protein LAMP-1 as intracellular entry factor. In endothelial cells expressing low levels of functional DG, both receptors are engaged by the virus and can contribute to productive entry. In sum, we characterize the role of Axl in LASV entry and provide a rationale to target Axl in anti-viral therapy.IMPORTANCEThe highly pathogenic arenavirus Lassa (LASV) represents a serious public health problem in Africa. Although the principal LASV receptor dystroglycan (DG) is ubiquitously expressed, virus binding critically depends on DG's post-translational modification, which does not always correlate with tissue tropism. The broadly expressed phosphatidylserine receptor Axl was recently identified as alternative LASV receptor candidate, but its role in LASV entry is unclear. Here we investigated the exact role of Axl in LASV entry as a function of DG's post-translational modification. We found that in absence of functional DG, Axl can mediate LASV entry via "apoptotic mimicry". Productive entry requires virus-induced receptor activation, involves macropinocytosis, and critically depends on LAMP-1. In endothelial cells that express low levels of glycosylated DG, both receptors can promote LASV entry. In sum, our study defines the roles of Axl in LASV entry and provides a rationale to target Axl in anti-viral therapy.oai:repository.helmholtz-hzi.de:10033/6213052019-08-30T11:26:13Zcom_10033_620636com_10033_620589col_10033_620665col_10033_620590
De Beuckelaer, Ans
Pollard, Charlotte
Van Lint, Sandra
Roose, Kenny
Van Hoecke, Lien
Naessens, Thomas
Udhayakumar, Vimal Kumar
Smet, Muriel
Sanders, Niek
Lienenklaus, Stefan
Saelens, Xavier
Weiss, Siegfried
Vanham, Guido
Grooten, Johan
De Koker, Stefaan
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
2018-03-06T09:44:42Z
2018-03-06T09:44:42Z
2016-11
Type I Interferons Interfere with the Capacity of mRNA Lipoplex Vaccines to Elicit Cytolytic T Cell Responses. 2016, 24 (11):2012-2020 Mol. Ther.
1525-0024
27506450
10.1038/mt.2016.161
http://hdl.handle.net/10033/621305
Molecular therapy : the journal of the American Society of Gene Therapy
Given their high potential to evoke cytolytic T cell responses, tumor antigen-encoding messenger RNA (mRNA) vaccines are now being intensively explored as therapeutic cancer vaccines. mRNA vaccines clearly benefit from wrapping the mRNA into nano-sized carriers such as lipoplexes that protect the mRNA from degradation and increase its uptake by dendritic cells in vivo. Nevertheless, the early innate host factors that regulate the induction of cytolytic T cells to mRNA lipoplex vaccines have remained unresolved. Here, we demonstrate that mRNA lipoplexes induce a potent type I interferon (IFN) response upon subcutaneous, intradermal and intranodal injection. Regardless of the route of immunization applied, these type I IFNs interfered with the generation of potent cytolytic T cell responses. Most importantly, blocking type I IFN signaling at the site of immunization through the use of an IFNAR blocking antibody greatly enhanced the prophylactic and therapeutic antitumor efficacy of mRNA lipoplexes in the highly aggressive B16 melanoma model. As type I IFN induction appears to be inherent to the mRNA itself rather than to unique properties of the mRNA lipoplex formulation, preventing type I IFN induction and/or IFNAR signaling at the site of immunization might constitute a widely applicable strategy to improve the potency of mRNA vaccination.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Animals
Antibodies
Cancer Vaccines
Humans
Injections, Intradermal
Injections, Subcutaneous
Interferon Type I
Liposomes
Melanoma, Experimental
Mice
RNA, Messenger
Receptor, Interferon alpha-beta
T-Lymphocytes, Cytotoxic
Treatment Outcome
Type I Interferons Interfere with the Capacity of mRNA Lipoplex Vaccines to Elicit Cytolytic T Cell Responses.
Article2018-06-13T21:40:17ZGiven their high potential to evoke cytolytic T cell responses, tumor antigen-encoding messenger RNA (mRNA) vaccines are now being intensively explored as therapeutic cancer vaccines. mRNA vaccines clearly benefit from wrapping the mRNA into nano-sized carriers such as lipoplexes that protect the mRNA from degradation and increase its uptake by dendritic cells in vivo. Nevertheless, the early innate host factors that regulate the induction of cytolytic T cells to mRNA lipoplex vaccines have remained unresolved. Here, we demonstrate that mRNA lipoplexes induce a potent type I interferon (IFN) response upon subcutaneous, intradermal and intranodal injection. Regardless of the route of immunization applied, these type I IFNs interfered with the generation of potent cytolytic T cell responses. Most importantly, blocking type I IFN signaling at the site of immunization through the use of an IFNAR blocking antibody greatly enhanced the prophylactic and therapeutic antitumor efficacy of mRNA lipoplexes in the highly aggressive B16 melanoma model. As type I IFN induction appears to be inherent to the mRNA itself rather than to unique properties of the mRNA lipoplex formulation, preventing type I IFN induction and/or IFNAR signaling at the site of immunization might constitute a widely applicable strategy to improve the potency of mRNA vaccination.oai:repository.helmholtz-hzi.de:10033/6213532020-01-08T14:12:14Zcom_10033_620589col_10033_620590
Baier, Claas
Haid, Sibylle
Beilken, Andreas
Behnert, Astrid
Wetzke, Martin
Brown, Richard J P
Schmitt, Corinna
Ebadi, Ella
Hansen, Gesine
Schulz, Thomas F
Pietschmann, Thomas
Bange, Franz-Christoph
TWINCORE, Zentrum für experimentelle uns klinische Ifektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2018-04-13T09:35:32Z
2018-04-13T09:35:32Z
2018
Molecular characteristics and successful management of a respiratory syncytial virus outbreak among pediatric patients with hemato-oncological disease. 2018, 7:21 Antimicrob Resist Infect Control
2047-2994
29449938
10.1186/s13756-018-0316-2
http://hdl.handle.net/10033/621353
Antimicrobial resistance and infection control
Respiratory syncytial virus (RSV) is responsible for upper and lower respiratory tract infection in adults and children. Especially immunocompromised patients are at high risk for a severe course of infection, and mortality is increased. Moreover RSV can spread in healthcare settings and can cause outbreaks. Herein we demonstrate the successful control and characteristics of a RSV outbreak that included 8 patients in our Department of Pediatric Hematology and Oncology.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Molecular characteristics and successful management of a respiratory syncytial virus outbreak among pediatric patients with hemato-oncological disease.
Article2018-06-13T04:25:00ZRespiratory syncytial virus (RSV) is responsible for upper and lower respiratory tract infection in adults and children. Especially immunocompromised patients are at high risk for a severe course of infection, and mortality is increased. Moreover RSV can spread in healthcare settings and can cause outbreaks. Herein we demonstrate the successful control and characteristics of a RSV outbreak that included 8 patients in our Department of Pediatric Hematology and Oncology.oai:repository.helmholtz-hzi.de:10033/6213692019-08-30T11:25:43Zcom_10033_620589col_10033_620590
Banse, Pia
Moeller, Rebecca
Bruening, Janina
Lasswitz, Lisa
Kahl, Sina
Khan, Abdul G
Marcotrigiano, Joseph
Pietschmann, Thomas
Gerold, Gisa
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2018-05-09T12:50:19Z
2018-05-09T12:50:19Z
2018
CD81 Receptor Regions outside the Large Extracellular Loop Determine Hepatitis C Virus Entry into Hepatoma Cells. 2018, 10 (4) Viruses
1999-4915
29677132
10.3390/v10040207
http://hdl.handle.net/10033/621369
Viruses
Hepatitis C virus (HCV) enters human hepatocytes using four essential entry factors, one of which is human CD81 (hCD81). The tetraspanin hCD81 contains a large extracellular loop (LEL), which interacts with the E2 glycoprotein of HCV. The role of the non-LEL regions of hCD81 (intracellular tails, four transmembrane domains, small extracellular loop and intracellular loop) is poorly understood. Here, we studied the contribution of these domains to HCV susceptibility of hepatoma cells by generating chimeras of related tetraspanins with the hCD81 LEL. Our results show that non-LEL regions in addition to the LEL determine susceptibility of cells to HCV. While closely related tetraspanins (X. tropicalis CD81 and D. rerio CD81) functionally complement hCD81 non-LEL regions, distantly related tetraspanins (C. elegans TSP9 amd D. melanogaster TSP96F) do not and tetraspanins with intermediate homology (hCD9) show an intermediate phenotype. Tetraspanin homology and susceptibility to HCV correlate positively. For some chimeras, infectivity correlates with surface expression. In contrast, the hCD9 chimera is fully surface expressed, binds HCV E2 glycoprotein but is impaired in HCV receptor function. We demonstrate that a cholesterol-coordinating glutamate residue in CD81, which hCD9 lacks, promotes HCV infection. This work highlights the hCD81 non-LEL regions as additional HCV susceptibility-determining factors.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
CD81 Receptor Regions outside the Large Extracellular Loop Determine Hepatitis C Virus Entry into Hepatoma Cells.
Article2018-06-13T05:38:34ZHepatitis C virus (HCV) enters human hepatocytes using four essential entry factors, one of which is human CD81 (hCD81). The tetraspanin hCD81 contains a large extracellular loop (LEL), which interacts with the E2 glycoprotein of HCV. The role of the non-LEL regions of hCD81 (intracellular tails, four transmembrane domains, small extracellular loop and intracellular loop) is poorly understood. Here, we studied the contribution of these domains to HCV susceptibility of hepatoma cells by generating chimeras of related tetraspanins with the hCD81 LEL. Our results show that non-LEL regions in addition to the LEL determine susceptibility of cells to HCV. While closely related tetraspanins (X. tropicalis CD81 and D. rerio CD81) functionally complement hCD81 non-LEL regions, distantly related tetraspanins (C. elegans TSP9 amd D. melanogaster TSP96F) do not and tetraspanins with intermediate homology (hCD9) show an intermediate phenotype. Tetraspanin homology and susceptibility to HCV correlate positively. For some chimeras, infectivity correlates with surface expression. In contrast, the hCD9 chimera is fully surface expressed, binds HCV E2 glycoprotein but is impaired in HCV receptor function. We demonstrate that a cholesterol-coordinating glutamate residue in CD81, which hCD9 lacks, promotes HCV infection. This work highlights the hCD81 non-LEL regions as additional HCV susceptibility-determining factors.oai:repository.helmholtz-hzi.de:10033/6214342019-08-30T11:34:19Zcom_10033_620589col_10033_620590
Lasswitz, Lisa
Chandra, Naresh
Arnberg, Niklas
Gerold, Gisa
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2018-08-02T08:12:33Z
2018-08-02T08:12:33Z
2018-06-22
1089-8638
29746851
10.1016/j.jmb.2018.04.039
http://hdl.handle.net/10033/621434
Adenoviruses as most viruses rely on glycan and protein interactions to attach to and enter susceptible host cells. The Adenoviridae family comprises more than 80 human types and they differ in their attachment factor and receptor usage, which likely contributes to the diverse tropism of the different types. In the past years, methods to systematically identify glycan and protein interactions have advanced. In particular sensitivity, speed and coverage of mass spectrometric analyses allow for high-throughput identification of glycans and peptides separated by liquid chromatography. Also, developments in glycan microarray technologies have led to targeted, high-throughput screening and identification of glycan-based receptors. The mapping of cell surface interactions of the diverse adenovirus types has implications for cell, tissue, and species tropism as well as drug development. Here we review known adenovirus interactions with glycan- and protein-based receptors, as well as glycomics and proteomics strategies to identify yet elusive virus receptors and attachment factors. We finally discuss challenges, bottlenecks, and future research directions in the field of non-enveloped virus entry into host cells.
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
adenovirus
glycomis
host cell interactions
proteomics
virus entry
Glycomics and Proteomics Approaches to Investigate Early Adenovirus-Host Cell Interactions.
Article
Journal of molecular biology
oai:repository.helmholtz-hzi.de:10033/6214832019-08-30T11:32:38Zcom_10033_620589col_10033_620590col_10033_620596
Behrendt, Patrick
Perin, Paula
Menzel, Nicolas
Branda, Dominic
Pfaender, Stephanie
Alves, Marco P.
Thiel, Volker
Meulemann, Philip
Colpit, Che C.
Schang, Luis M.
Vondran, Florian W.R.
Anggakusuma
Manns, Michael P.
Steinmann, Eicke
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2018-09-17T11:40:52Z
2018-09-17T11:40:52Z
2017-01-01
1872-9096
28923507
10.1016/j.antiviral.2017.09.006
http://hdl.handle.net/10033/621483
Approximately 142 million people worldwide are infected with hepatitis C virus (HCV). Although potent direct acting antivirals are available, high costs limit access to treatment. Chronic hepatitis C virus infection remains a major cause of orthotopic liver transplantation. Moreover, re-infection of the graft occurs regularly. Antivirals derived from natural sources might be an alternative and cost-effective option to complement therapy regimens for global control of hepatitis C virus infection. We tested the antiviral properties of a mixture of different Chinese herbs/roots named Zhi Bai Di Huang Wan (ZBDHW) and its individual components on HCV. One of the ZBDHW components, Penta-O-Galloyl-Glucose (PGG), was further analyzed for its mode of action in vitro, its antiviral activity in primary human hepatocytes as well as for its bioavailability and hepatotoxicity in mice. ZBDHW, its component Cortex Moutan and the compound PGG efficiently block entry of HCV of all major genotypes and also of the related flavivirus Zika virus. PGG does not disrupt HCV virion integrity and acts primarily during virus attachment. PGG shows an additive effect when combined with the well characterized HCV inhibitor Daclatasvir. Analysis of bioavailability in mice revealed plasma levels above tissue culture IC
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Antivirals
Bioavailability
Cortex moutan
Entry inhibitor
Hepatitis C virus
Natural compounds
Penta-O-Galloyl-Glucose
Pentagalloylglucose, a highly bioavailable polyphenolic compound present in Cortex moutan, efficiently blocks hepatitis C virus entry.
Article
Antiviral research2018-09-18T00:00:00Zoai:repository.helmholtz-hzi.de:10033/6214812019-08-30T11:29:14Zcom_10033_620589col_10033_620590
Bruening, Janina
Lasswitz, Lisa
Banse, Pia
Kahl, Sina
Marinach, Carine
Vondran, Florian W
Kaderali, Lars
Silvie, Olivier
Pietschmann, Thomas
Meissner, Felix
Gerold, Gisa
2018-09-17T07:08:53Z
2018-09-17T07:08:53Z
2018-07-01
1553-7374
30024968
10.1371/journal.ppat.1007111
http://hdl.handle.net/10033/621481
Hepatitis C virus (HCV) and the malaria parasite Plasmodium use the membrane protein CD81 to invade human liver cells. Here we mapped 33 host protein interactions of CD81 in primary human liver and hepatoma cells using high-resolution quantitative proteomics. In the CD81 protein network, we identified five proteins which are HCV entry factors or facilitators including epidermal growth factor receptor (EGFR). Notably, we discovered calpain-5 (CAPN5) and the ubiquitin ligase Casitas B-lineage lymphoma proto-oncogene B (CBLB) to form a complex with CD81 and support HCV entry. CAPN5 and CBLB were required for a post-binding and pre-replication step in the HCV life cycle. Knockout of CAPN5 and CBLB reduced susceptibility to all tested HCV genotypes, but not to other enveloped viruses such as vesicular stomatitis virus and human coronavirus. Furthermore, Plasmodium sporozoites relied on a distinct set of CD81 interaction partners for liver cell entry. Our findings reveal a comprehensive CD81 network in human liver cells and show that HCV and Plasmodium highjack selective CD81 interactions, including CAPN5 and CBLB for HCV, to invade cells.
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Hepatitis C virus enters liver cells using the CD81 receptor complex proteins calpain-5 and CBLB.
Article
PLoS pathogens2018-09-17T07:08:53Zoai:repository.helmholtz-hzi.de:10033/6215182019-08-30T11:29:45Zcom_10033_620589col_10033_620590col_10033_620596
Todt, Daniel
Moeller, Nora
Praditya, Dimas
Kinast, Volker
Friesland, Martina
Engelmann, Michael
Verhoye, Lieven
Sayed, Ibrahim M
Behrendt, Patrick
Dao Thi, Viet Loan
Meuleman, Philip
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2018-10-12T10:49:42Z
2018-10-12T10:49:42Z
2018-09-01
1872-9096
30036559
10.1016/j.antiviral.2018.07.010
http://hdl.handle.net/10033/621518
Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and a member of the genus Orthohepevirus in the family Hepeviridae. HEV infections are the common cause of acute hepatitis but can also take chronic courses. Ribavirin is the treatment of choice for most patients and type I interferon (IFN) has been evaluated in a few infected transplantation patients in vivo. However, no effective and specific treatments against HEV infections are currently available. In this study, we evaluated the natural compound silvestrol, isolated from the plant Aglaia foveolata, and known for its specific inhibition of the DEAD-box RNA helicase eIF4A in state-of-the-art HEV experimental model systems. Silvestrol blocked HEV replication of different subgenomic replicons in a dose-dependent manner at low nanomolar concentrations and acted additive to ribavirin (RBV). In addition, HEV p6-based full length replication and production of infectious particles was reduced in the presence of silvestrol. A pangenotypic effect of the compound was further demonstrated with primary isolates from four different human genotypes in HEV infection experiments of hepatocyte-like cells derived from human embryonic and induced pluripotent stem cells. In vivo, HEV RNA levels rapidly declined in the feces of treated mice while no effect was observed in the vehicle treated control animals. In conclusion, silvestrol could be identified as pangenotypic HEV replication inhibitor in vitro with additive effect to RBV and further demonstrated high potency in vivo. The compound therefore may be considered in future treatment strategies of chronic hepatitis E in immunocompromised patients.
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Antiviral activity
Hepatitis E virus (HEV)
Host target
Humanized mice
Replication
Silvestrol
The natural compound silvestrol inhibits hepatitis E virus (HEV) replication in vitro and in vivo.
Article
Antiviral research2018-10-12T10:49:43Zoai:repository.helmholtz-hzi.de:10033/6215492019-08-30T11:32:11Zcom_10033_620589col_10033_620590
García-Nicolás, Obdulio
V'kovski, Philip
Vielle, Nathalie J
Ebert, Nadine
Züst, Roland
Portmann, Jasmine
Stalder, Hanspeter
Gaschen, Véronique
Vieyres, Gabrielle
Stoffel, Michael
Schweizer, Matthias
Summerfield, Artur
Engler, Olivier
Pietschmann, Thomas
Todt, Daniel
Alves, Marco P
Thiel, Volker
Pfaender, Stephanie
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2018-11-09T10:16:10Z
2018-11-09T10:16:10Z
2018-11-01
1098-6596
30181371
10.1128/AAC.01206-18
http://hdl.handle.net/10033/621549
The virus family
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Flaviviridae
K22
antiviral
flavivirus
hepacivirus
panviral inhibitor
pestivirus
The Small-Compound Inhibitor K22 Displays Broad Antiviral Activity against Different Members of the Family Flaviviridae and Offers Potential as a Panviral Inhibitor.
Article
Antimicrobial agents and chemotherapy2018-11-13T10:40:30Zoai:repository.helmholtz-hzi.de:10033/6216062019-08-30T11:31:43Zcom_10033_620589col_10033_620590
Abere, Bizunesh
Samarina, Naira
Gramolelli, Silvia
Rückert, Jessica
Gerold, Gisa
Pich, Andreas
Schulz, Thomas F
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2018-12-05T13:45:30Z
2018-12-05T13:45:30Z
2018-09-01
1098-5514
29950425
10.1128/JVI.00544-18
http://hdl.handle.net/10033/621606
Kaposi's sarcoma (KS)-associated herpesvirus (KSHV)/human herpesvirus 8 (HHV-8) causes the angiogenic tumor KS and two B-cell malignancies. The KSHV nonstructural membrane protein encoded by the open reading frame (ORF) K15 recruits and activates several cellular proteins, including phospholipase Cγ1 (PLCγ1), components of the NF-κB pathway, as well as members of the Src family of nonreceptor tyrosine kinases, and thereby plays an important role in the activation of angiogenic and inflammatory pathways that contribute to the pathogenesis of KS as well as KSHV productive (lytic) replication. In order to identify novel cellular components involved in the biology of pK15, we immunoprecipitated pK15 from KSHV-infected endothelial cells and identified associated proteins by label-free quantitative mass spectrometry. Cellular proteins interacting with pK15 point to previously unappreciated cellular processes, such as the endocytic pathway, that could be involved in the function of pK15. We found that the class II phosphatidylinositol 3-kinase (PI3K) PI3K-C2α, which is involved in the endocytosis of activated receptor tyrosine kinases and their signaling from intracellular organelles, interacts and colocalizes with pK15 in vesicular structures abundant in the perinuclear area. Further functional analysis revealed that PI3K-C2α contributes to the pK15-dependent phosphorylation of PLCγ1 and Erk1/2. PI3K-C2α also plays a role in KSHV lytic replication, as evidenced by the reduced expression of the viral lytic genes K-bZIP and ORF45 as well as the reduced release of infectious virus in PI3K-C2α-depleted KSHV-infected endothelial cells. Taken together, our results suggest a role of the cellular PI3K-C2α protein in the functional properties of the KSHV pK15 protein.
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
KSHV-K15
PI3K-C2α
lytic replication
Kaposi's Sarcoma-Associated Herpesvirus Nonstructural Membrane Protein pK15 Recruits the Class II Phosphatidylinositol 3-Kinase PI3K-C2α To Activate Productive Viral Replication.
Article
Journal of virology
oai:repository.helmholtz-hzi.de:10033/6216522019-08-30T11:31:21Zcom_10033_620589col_10033_620590
Vasiliauskaite, Ieva
Owsianka, Ania
England, Patrick
Khan, Abdul Ghafoor
Cole, Sarah
Bankwitz, Dorothea
Foung, Steven K H
Pietschmann, Thomas
Marcotrigiano, Joseph
Rey, Felix A
Patel, Arvind H
Krey, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-01-16T09:27:48Z
2019-01-16T09:27:48Z
2017-05-16
MBio. 2017 May 16;8(3). pii: mBio.00382-17. doi: 10.1128/mBio.00382-17.
2150-7511
28512091
10.1128/mBio.00382-17
http://hdl.handle.net/10033/621652
The hepatitis C virus (HCV) glycoprotein E2 is the major target of neutralizing antibodies and is therefore highly relevant for vaccine design. Its structure features a central immunoglobulin (Ig)-like β-sandwich that contributes to the binding site for the cellular receptor CD81. We show that a synthetic peptide corresponding to a β-strand of this Ig-like domain forms an α-helix in complex with the anti-E2 antibody DAO5, demonstrating an inside-out flip of hydrophobic residues and a secondary structure change in the composite CD81 binding site. A detailed interaction analysis of DAO5 and cross-competing neutralizing antibodies with soluble E2 revealed that the Ig-like domain is trapped by different antibodies in at least two distinct conformations. DAO5 specifically captures retrovirus particles bearing HCV glycoproteins (HCVpp) and infectious cell culture-derived HCV particles (HCVcc). Infection of cells by DAO5-captured HCVpp can be blocked by a cross-competing neutralizing antibody, indicating that a single virus particle simultaneously displays E2 molecules in more than one conformation on its surface. Such conformational plasticity of the HCV E2 receptor binding site has important implications for immunogen design.
ASM
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
CD81 binding site
Ig-like domain
conformational flexibility
glycoprotein E2
hepatitis C virus
monoclonal antibodies
vaccine design
Conformational Flexibility in the Immunoglobulin-Like Domain of the Hepatitis C Virus Glycoprotein E2.
Article
mBio2019-01-16T09:27:49Zoai:repository.helmholtz-hzi.de:10033/6216982019-08-30T11:32:39Zcom_10033_620589col_10033_620590
Pietschmann, Thomas
Brown, Richard J P
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-02-19T13:32:33Z
2019-02-19T13:32:33Z
2019-01-29
1878-4380
30709707
10.1016/j.tim.2019.01.001
http://hdl.handle.net/10033/621698
Trends in Microbiology
Hepatitis C virus (HCV) is an enveloped, RNA virus transmitted through blood-to-blood contact. It infects humans only and primarily targets liver cells. HCV evades innate and adaptive immunity and establishes chronic infections in 70% of cases. If untreated, 20% of patients develop liver cirrhosis, and a fraction of these progress to hepatocellular carcinoma. Annually, 400000 patients die globally due to HCV infection. Direct-acting antivirals (DAAs) are licensed and target three viral proteins: the NS3-4A protease needed for processing the viral polyprotein, the NS5A phosphoprotein that regulates RNA replication and virus assembly, and the viral RNA-dependent RNA polymerase (NS5B) that catalyzes genome replication. Combination therapies cure more than 95% of treated patients. Approximately 71 million people are chronically infected and 1.7 million new infections occur annually. Treatment-induced cure does not protect from viral reinfection. A prophylactic vaccine is under development.
Elsevier
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
HCV
antiviral therapy
infection
liver disease
Hepatitis C Virus.
Article
Trends in microbiology
oai:repository.helmholtz-hzi.de:10033/6217452019-08-30T11:31:43Zcom_10033_620589col_10033_620590col_10033_620596
Behrendt, Patrick
Brüning, Janina
Todt, Daniel
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str.7,30625 Hannover, Germany.
2019-04-12T08:54:44Z
2019-04-12T08:54:44Z
2019-03-01
Open Forum Infect Dis. 2019 Feb 13;6(3):ofz047. doi: 10.1093/ofid/ofz047. eCollection 2019 Mar.
2328-8957
30882013
10.1093/ofid/ofz047
http://hdl.handle.net/10033/621745
Open Forum Infectious diseases
Hepatitis C virus (HCV) is a blood-borne virus and is most frequently transmitted through large or repeated direct percutaneous exposures to infected blood. The 2 most common exposures associated with transmission of HCV are blood transfusion and intravenous drug abuse. The association between HCV transmission and other suspected risk factors such as tattooing is more controversial. Although HCV can survive for days to weeks in suspension or on inanimate surfaces, its stability in tattooing supplies remains elusive. Here, we analyzed the influence of tattoo ink on HCV infectiousness.
en
Oxford Academic
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
hepatitis C virus
prevention
transmission
viral stability
Influence of Tattoo Ink on Hepatitis C Virus Infectiousness.
Article
Open forum infectious diseases2019-04-12T08:54:44Zoai:repository.helmholtz-hzi.de:10033/6217552021-03-03T11:16:24Zcom_10033_620589col_10033_620590
Martin-Benlloch, Xavier
Haid, Sybille
Novodomska, Alexandra
Rominger, Frank
Pietschmann, Thomas
Davioud-Charvet, Elisabeth
Elhabiri, Mourad
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str.7,30625 Hannover, Germany.
2019-04-18T13:16:05Z
2019-04-18T13:16:05Z
2019-03-05
ACS Omega 2019 Mar 5;4(3):4871. doi: 10.1021/acsomega.8b03332.
2470-1343
31459671
10.1021/acsomega.8b03332
http://hdl.handle.net/10033/621755
ACS Omega
Ladanein (i.e., 5,6,7-trihydroxylated flavone) was demonstrated to act as a powerful virucidal agent toward a broad range of enveloped virus particles. Fe(III) coordination and pH are indeed among the key parameters that might favor both bioactivation of the flavone and consequent host cell entry inhibition. In this present work, the impact of fluorinated groups on the physicochemical and antiviral properties of the flavone was investigated, thus allowing a deeper understanding of the antiviral mode of action. The improved synthesis of ladanein allowed accessing a broad range of analogues, some of them being significantly more active than the former ladanein lead compound. We first determined the acido-basic properties of this homogenous series of compounds and then investigated their electrochemical behavior. Fe(III) coordination properties (stability, spectral behavior, and kinetics) of ladanein and its analogues were then examined (quasiphysiological conditions) and provided key information of their stability and reactivity. Using the determined physicochemical parameters, the critical impact of the iron complexation and medium acidity was confirmed on hepatitis C virus (HCV) particles (pre)treated with ladanein. Finally, a preliminary structure–HCV entry inhibition relationship study evidenced the superior antiviral activity of the ladanein analogues bearing an electron-withdrawing group in para position (FCF3 > FOCF3 > FFCF3 > FF > FOMe) on the B cycle in comparison with the parent ladanein itself.
en
American Chemical Society
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Physicochemical Properties Govern the Activity of Potent Antiviral Flavones
Article
oai:repository.helmholtz-hzi.de:10033/6217662019-08-30T11:35:08Zcom_10033_620589col_10033_620590
Weller, Romy
Hueging, Kathrin
Brown, Richard J P
Todt, Daniel
Joecks, Sebastian
Vondran, Florian W R
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str.7,30625 Hannover, Germany.
2019-05-09T12:48:57Z
2019-05-09T12:48:57Z
2017-09-15
J Virol. 2017 Aug 24;91(18). pii: JVI.00422-17. doi: 10.1128/JVI.00422-17. Print 2017 Sep 15.
1098-5514
28659481
10.1128/JVI.00422-17
http://hdl.handle.net/10033/621766
Journal of Virology
Hepatitis C virus (HCV) is extraordinarily diverse and uses entry factors in a strain-specific manner. Virus particles associate with lipoproteins, and apolipoprotein E (ApoE) is critical for HCV assembly and infectivity. However, whether ApoE dependency is common to all HCV genotypes remains unknown. Therefore, we compared the roles of ApoE utilizing 10 virus strains from genotypes 1 through 7. ApoA and ApoC also support HCV assembly, so they may contribute to virus production in a strain-dependent fashion. Transcriptome sequencing (RNA-seq) revealed abundant coexpression of ApoE, ApoB, ApoA1, ApoA2, ApoC1, ApoC2, and ApoC3 in primary hepatocytes and in Huh-7.5 cells. Virus production was examined in Huh-7.5 cells with and without ApoE expression and in 293T cells where individual apolipoproteins (ApoE1, -E2, -E3, -A1, -A2, -C1, and -C3) were provided in trans All strains were strictly ApoE dependent. However, ApoE involvement in virus production was strain and cell type specific, because some HCV strains poorly produced infectious virus in ApoE-expressing 293T cells and because ApoE knockout differentially affected virus production of HCV strains in Huh-7.5 cells. ApoE allelic isoforms (ApoE2, -E3, and -E4) complemented virus production of HCV strains to comparable degrees. All tested strains assembled infectious progeny with ApoE in preference to other exchangeable apolipoproteins (ApoA1, -A2, -C1, and -C3). The specific infectivity of HCV particles was similar for 293T- and Huh-7.5-derived particles for most strains; however, it differed by more than 100-fold in some viruses. Collectively, this study reveals strain-dependent and host cell-dependent use of ApoE during HCV assembly. These differences relate to the efficacy of virus production and also to the properties of released virus particles and therefore govern viral fitness at the level of assembly and cell entry.IMPORTANCE Chronic HCV infections are a major cause of liver disease. HCV is highly variable, and strain-specific determinants modulate the response to antiviral therapy, the natural course of infection, and cell entry factor usage. Here we explored whether host factor dependency of HCV in particle assembly is modulated by strain-dependent viral properties. We showed that all examined HCV strains, which represent all seven known genotypes, rely on ApoE expression for assembly of infectious progeny. However, the degree of ApoE dependence is modulated in a strain-specific and cell type-dependent manner. This indicates that HCV strains differ in their assembly properties and host factor usage during assembly of infectious progeny. Importantly, these differences relate not only to the efficiency of virus production and release but also to the infectiousness of virus particles. Thus, strain-dependent features of HCV modulate ApoE usage, with implications for virus fitness at the level of assembly and cell entry.
en
American Society for Microbiology
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
ApoE
apolipoprotein
assembly
genotypes
hepatitis C virus
Hepatitis C Virus Strain-Dependent Usage of Apolipoprotein E Modulates Assembly Efficiency and Specific Infectivity of Secreted Virions.
Article
Journal of virology
oai:repository.helmholtz-hzi.de:10033/6217882019-08-30T11:35:09Zcom_10033_620589col_10033_620590
Vieyres, Gabrielle
Pietschmann, Thomas
TWINCORE, Zentrum für Experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2019-05-21T09:35:35Z
2019-05-21T09:35:35Z
2019-03-12
Cells. 2019 Mar 12;8(3). pii: cells8030233. doi: 10.3390/cells8030233.
2073-4409
30871009
10.3390/cells8030233
http://hdl.handle.net/10033/621788
Cells
The replication cycle of the liver-tropic hepatitis C virus (HCV) is tightly connected to the host lipid metabolism, during the virus entry, replication, assembly and egress stages, but also while the virus circulates in the bloodstream. This interplay coins viral particle properties, governs viral cell tropism, and facilitates immune evasion. This review summarizes our knowledge of these interactions focusing on the late steps of the virus replication cycle. It builds on our understanding of the cell biology of lipid droplets and the biosynthesis of liver lipoproteins and attempts to explain how HCV hijacks these organelles and pathways to assemble its lipo-viro-particles. In particular, this review describes (i) the mechanisms of viral protein translocation to and from the lipid droplet surface and the orchestration of an interface between replication and assembly complexes, (ii) the importance of the triglyceride mobilization from the lipid droplets for HCV assembly, (iii) the interplay between HCV and the lipoprotein synthesis pathway including the role played by apolipoproteins in virion assembly, and finally (iv) the consequences of these complex virus–host interactions on the virion composition and its biophysical properties. The wealth of data accumulated in the past years on the role of the lipid metabolism in HCV assembly and its imprint on the virion properties will guide vaccine design efforts and reinforce our understanding of the hepatic lipid metabolism in health and disease.
en
MPDI
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
DGAT
VLDL
apolipoprotein
hepatitis C virus
lipid droplet
lipolysis
lipoprotein
membrane contact sites
molecular mimicry
virus assembly and release
HCV Pit Stop at the Lipid Droplet: Refuel Lipids and Put on a Lipoprotein Coat before Exit.
Article
Cells
oai:repository.helmholtz-hzi.de:10033/6217992019-08-30T11:30:27Zcom_10033_620857com_10033_620589col_10033_620858col_10033_620590
Sandargo, Birthe
Michehl, Maira
Praditya, Dimas
Steinmann, Eike
Stadler, Marc
Surup, Frank
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany; TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-06-05T08:00:25Z
2019-06-05T08:00:25Z
2019-05-03
Org Lett. 2019 May 3;21(9):3286-3289. doi: 10.1021/acs.orglett.9b01017. Epub 2019 Apr 22.
1523-7052
31008606
10.1021/acs.orglett.9b01017
http://hdl.handle.net/10033/621799
Organic Letters
Rhodatin (1), a meroterpenoid featuring a unique pentacyclic scaffold with both spiro and spiroketal centers, and five unusual acorane-type sesquiterpenoids, named rhodocoranes A-E (2-6, respectively), are the first natural products isolated from the basidiomycete Rhodotus palmatus. Their structures were elucidated by two-dimensional NMR experiments and HRESIMS, while the absolute configuration of the substance family was determined by Mosher's method utilizing 2. Rhodatin strongly inhibited hepatitis C virus, whereas 4 displayed cytotoxicity and selective antifungal activity.
en
American Chemical Society
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Antiviral Meroterpenoid Rhodatin and Sesquiterpenoids Rhodocoranes A-E from the Wrinkled Peach Mushroom, Rhodotus palmatus.
Article
Organic letters
oai:repository.helmholtz-hzi.de:10033/6218022019-08-30T11:33:01Zcom_10033_620589com_10033_620618col_10033_620619col_10033_620590
Banda, Dominic H
Perin, Paula M
Brown, Richard J P
Todt, Daniel
Solodenko, Wladimir
Hoffmeyer, Patrick
Kumar Sahu, Kamlesh
Houghton, Michael
Meuleman, Philip
Müller, Rolf
Kirschning, Andreas
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-06-06T12:50:11Z
2019-06-06T12:50:11Z
2019-06-01
J Hepatol. 2019 Jun;70(6):1082-1092. doi: 10.1016/j.jhep.2019.01.033. Epub 2019 Feb 13.
1600-0641
30769006
10.1016/j.jhep.2019.01.033
http://hdl.handle.net/10033/621802
Journal of Hepatology
en
Elsevier
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Antivirals
Fusion inhibitors
Hepatitis C virus (HCV)
Membrane fusion
Resistance
A central hydrophobic E1 region controls the pH range of hepatitis C virus membrane fusion and susceptibility to fusion inhibitors.
Article
Journal of hepatology
oai:repository.helmholtz-hzi.de:10033/6218412019-11-21T11:57:09Zcom_10033_620589col_10033_620590
Meister, Toni Luise
Tegtmeyer, Birthe
Postel, Alexander
Cavalleri, Jessika-M V
Todt, Daniel
Stang, Alexander
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-07-02T13:16:22Z
2019-07-02T13:16:22Z
2019-05-21
Viruses. 2019 May 21;11(5). pii: v11050461. doi: 10.3390/v11050461.
1999-4915
31117220
10.3390/v11050461
http://hdl.handle.net/10033/621841
Viruses
An equine parvovirus-hepatitis (EqPV-H) has been recently identified in association with equine serum hepatitis, also known as Theiler's disease. This disease was first described by Arnold Theiler in 1918 and is often observed after applications with blood products in equines. So far, the virus has only been described in the USA and China. In this study, we evaluated the presence of EqPV-H in several commercial serum samples to assess the potential risk of virus transmission by equine serum-based products for medical and research applications. In 11 out of 18 commercial serum samples, EqPV-H DNA was detectable with a viral load up to 105 copies/mL. The same serum batches as well as three additional samples were also positive for antibodies against the EqPV-H VP1 protein. The countries of origin with detectable viral genomes included the USA, Canada, New Zealand, Italy, and Germany, suggesting a worldwide distribution of EqPV-H. Phylogenetic analysis of the EqPV-H NS1 sequence in commercial serum samples revealed high similarities in viral sequences from different geographical areas. As horse sera are commonly used for the production of anti-sera, which are included in human and veterinary medical products, these results implicate the requirement for diagnostic tests to prevent EqPV-H transmission.
en
MPDI
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
commercial horse serum
equine parvovirus-hepatitis
horses
phylogeny
Equine Parvovirus-Hepatitis Frequently Detectable in Commercial Equine Serum Pools.
Article
Viruses
oai:repository.helmholtz-hzi.de:10033/6218872019-08-30T11:26:41Zcom_10033_620589col_10033_620590
Kinast, Volker
Leber, Stefan L
Brown, Richard J P
Vieyres, Gabrielle
Behrendt, Patrick
Eßbach, Constanze
Strnad, Pavel
Vondran, Florian W R
Cornberg, Markus
Wex, Cora
Pietschmann, Thomas
Haybaeck, Johannes
Todt, Daniel
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-07-23T09:00:36Z
2019-07-23T09:00:36Z
2019-06-18
Cells. 2019 Jun 18;8(6). pii: cells8060610. doi: 10.3390/cells8060610.
2073-4409
31216713
10.3390/cells8060610
http://hdl.handle.net/10033/621887
Cells
Keratin proteins form intermediate filaments, which provide structural support for many tissues. Multiple keratin family members are reported to be associated with the progression of liver disease of multiple etiologies. For example, keratin 23 (KRT23) was reported as a stress-inducible protein, whose expression levels correlate with the severity of liver disease. Hepatitis C virus (HCV) is a human pathogen that causes chronic liver diseases including fibrosis, cirrhosis, and hepatocellular carcinoma. However, a link between KRT23 and hepatitis C virus (HCV) infection has not been reported previously. In this study, we investigated KRT23 mRNA levels in datasets from liver biopsies of chronic hepatitis C (CHC) patients and in primary human hepatocytes experimentally infected with HCV, in addition to hepatoma cells. Interestingly, in each of these specimens, we observed an HCV-dependent increase of mRNA levels. Importantly, the KRT23 protein levels in patient plasma decreased upon viral clearance. Ectopic expression of KRT23 enhanced HCV infection; however, CRIPSPR/Cas9-mediated knockout did not show altered replication efficiency. Taken together, our study identifies KRT23 as a novel, virus-induced host-factor for hepatitis C virus.
MPDI
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
hepatitis C virus (HCV)
host factor
keratin 23
virus–host interaction
Identification of Keratin 23 as a Hepatitis C Virus-Induced Host Factor in the Human Liver.
Article
Cells
oai:repository.helmholtz-hzi.de:10033/6219052019-08-30T11:26:11Zcom_10033_620589col_10033_620590
Praditya, Dimas
Kirchhoff, Lisa
Brüning, Janina
Rachmawati, Heni
Steinmann, Joerg
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-08-16T08:53:57Z
2019-08-16T08:53:57Z
2019-01-01
Front Microbiol. 2019 May 3;10:912. doi: 10.3389/fmicb.2019.00912. eCollection 2019.
1664-302X
31130924
10.3389/fmicb.2019.00912
http://hdl.handle.net/10033/621905
Frontiers in Microbiology
The search for novel anti-infectives is one of the most important challenges in natural product research, as diseases caused by bacteria, viruses, and fungi are influencing the human society all over the world. Natural compounds are a continuing source of novel anti-infectives. Accordingly, curcumin, has been used for centuries in Asian traditional medicine to treat various disorders. Numerous studies have shown that curcumin possesses a wide spectrum of biological and pharmacological properties, acting, for example, as anti-inflammatory, anti-angiogenic and anti-neoplastic, while no toxicity is associated with the compound. Recently, curcumin's antiviral and antibacterial activity was investigated, and it was shown to act against various important human pathogens like the influenza virus, hepatitis C virus, HIV and strains of Staphylococcus, Streptococcus, and Pseudomonas. Despite the potency, curcumin has not yet been approved as a therapeutic antiviral agent. This review summarizes the current knowledge and future perspectives of the antiviral, antibacterial, and antifungal effects of curcumin.
en
Frontiers
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
anti-infective properties
bacteria
curcumin
fungi
natural products
nutraceutical
virus
Anti-infective Properties of the Golden Spice Curcumin.
Article
Frontiers in microbiology
oai:repository.helmholtz-hzi.de:10033/6219502021-04-16T12:27:58Zcom_10033_620589col_10033_620590
Tegtmeyer, Birthe
Echelmeyer, Julia
Pfankuche, Vanessa M
Puff, Christina
Todt, Daniel
Fischer, Nicole
Durham, Andy
Feige, Karsten
Baumgärtner, Wolfgang
Steinmann, Eike
Cavalleri, Jessika-M V
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-09-18T13:03:42Z
2019-09-18T13:03:42Z
2019-01-01
Vet Med Sci. 2019 Aug;5(3):372-378. doi: 10.1002/vms3.181. Epub 2019 Jul 3.
2053-1095
31267690
10.1002/vms3.181
http://hdl.handle.net/10033/621950
Veterinary Medicine and Science
Background: Equine hepacivirus (EqHV) in equids represents the closest homologue to hepatitis C virus(HCV) infecting humans. A majority of HCV infected patients develop a chronic course of infection leading toliver fibrosis, cirrhosis and liver failure. However, in horses mostly transient mild subclinical infections arereported for EqHV to date. Objectives: EqHV can be involved in chronic liver diseases of horses. Methods:Biochemical parameters in serum samples were measured. Viral load was determined using qPCR. Next gener-ation sequencing (NGS) of serum was performed. Liver tissue was stained with haematoxylin and eosin andanalysed for viral RNA with fluorescentin situ-hybridization. Results: The horse showed symptoms of severehepatopathy and was chronically infected with EqHV. Viral RNA was detectable in the liver during disease.To rule out other infectious agents NGS was performed and showed the highest abundance for EqHV. Theidentified virus sequence was similar to other circulating equine hepaciviruses. Conclusions: EqHV can be asso-ciated with liver disease in horses. Whether it causes the disease or contributes in a multifactorial mannerneeds further investigation
en
Wiley Open Access
info:eu-repo/grantAgreement/EC/H2020/ 643476
openAccess
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc/4.0/
equine hepacivirus
hepacivirus A
hepatitis C virus
hepatopathy
liver
Chronic equine hepacivirus infection in an adult gelding with severe hepatopathy.
Article
Veterinary medicine and science
oai:repository.helmholtz-hzi.de:10033/6219622021-01-19T03:37:34Zcom_10033_620589col_10033_620590
Schlevogt, Bernhard
Kinast, Volker
Reusch, Julia
Kerkhoff, Andrea
Praditya, Dimas
Todt, Daniel
Schmidt, Hartmut H
Steinmann, Eike
Behrendt, Patrick
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-09-24T14:02:31Z
2019-09-24T14:02:31Z
2019-08-22
Pathogens. 2019 Aug 22;8(3). pii: pathogens8030129. doi: 10.3390/pathogens8030129.
2076-0817
31443360
10.3390/pathogens8030129
http://hdl.handle.net/10033/621962
Pathogens
Hepatitis E virus (HEV) is an increasingly recognised pathogen, affecting several hundred thousand individuals in western countries each year. Importantly, the majority of immunocompromised individuals are not able to clear HEV but develop a chronic course of infection. In the case of lymphoma, which is an inherent immunosuppressive disease per se, chemotherapy can even further exacerbate the immunosuppressive status. As the mechanism of HEV chronification is barely understood, it is important to gain knowledge about the influence of chemotherapeutic drugs on the HEV replication cycle to guide rational clinical management of HEV infection in such patients. In this case report, a 70 year old man was diagnosed with lymphoplasmacytic lymphoma. As we observed the occurrence of chronic HEV after treatment with the Bruton's tyrosine kinase (BTK) inhibitor ibrutinib in vivo, we investigated the influence of BTK signaling and ibrutinib treatment in the HEV replication cycle in vitro. First, we detected an HEV-induced mobilisation of BTK in human liver cells during HEV replication. A moderate antiviral effect against HEV replicating isolates including genotypes 1 and 3 was observed, suggesting that ibrutinib did not support HEV replication in a direct manner. Combinatory treatments of ibrutinib with ribavirin indicated that ibrutinib did not influence the antiviral effect of ribavirin. Taken together, chemotherapy targeting cellular factors for the treatment of lymphomas may be a neglected risk factor for the chronification of HEV. For ibrutinib, despite the upregulation of its target BTK during HEV replication, we observed neither a proviral effect on HEV replication nor an influence on the antiviral effect of ribavirin, suggesting that the chronification of HEV may be favoured by its immunosuppressive effect.
en
MPDI
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Bruton’s tyrosine kinase
chronification
hepatitis E virus
ibrutinib
immunosuppression
lymphoplasmacytic lymphoma
Chronic Hepatitis E Virus Infection during Lymphoplasmacytic Lymphoma and Ibrutinib Treatment.
Article
Pathogens (Basel, Switzerland)
oai:repository.helmholtz-hzi.de:10033/6219762019-12-07T02:08:41Zcom_10033_620589col_10033_620590
Köllmann, Christoph
Wiechert, Svenja M.
Jones, Peter G.
Pietschmann, Thomas
Werz, Daniel B.
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Köllmann, C.
2019-10-14T14:36:29Z
2019-10-14T14:36:29Z
2019-09-06
Org Lett. 2019 Aug 21. doi: 10.1021/acs.orglett.9b02555.
15237060
31433193
10.1021/acs.orglett.9b02555
https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85071948253&origin=inward
http://hdl.handle.net/10033/621976
Organic Letters
2-s2.0-85071948253
SCOPUS_ID:85071948253
American Chemical Society
Organic Letters
17
21
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Synthesis of 4′/5′-Spirocyclopropanated Uridine and d -Xylouridine Derivatives and Their Activity against the Human Respiratory Syncytial Virus
Article
oai:repository.helmholtz-hzi.de:10033/6220032019-11-21T12:46:36Zcom_10033_620589col_10033_620590
Meister, Toni Luise
Tegtmeyer, Birthe
Brüggemann, Yannick
Sieme, Harald
Feige, Karsten
Todt, Daniel
Stang, Alexander
Cavalleri, Jessika-M V
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-11-05T14:51:16Z
2019-11-05T14:51:16Z
2019-10-18
Viruses. 2019 Oct 18;11(10). pii: v11100965. doi: 10.3390/v11100965.
1999-4915
31635426
10.3390/v11100965
http://hdl.handle.net/10033/622003
Viruses
en
MPDI
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Germany
equine parvovirus-hepatitis
risk factors
transmission
Characterization of Equine Parvovirus in Thoroughbred Breeding Horses from Germany.
Article
Viruses
oai:repository.helmholtz-hzi.de:10033/6220042019-11-06T02:19:46Zcom_10033_620589col_10033_620590
Kleinert, Robin D V
Montoya-Diaz, Eduardo
Khera, Tanvi
Welsch, Kathrin
Tegtmeyer, Birthe
Hoehl, Sebastian
Ciesek, Sandra
Brown, Richard J P
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-11-05T15:06:29Z
2019-11-05T15:06:29Z
2019-10-17
Viruses. 2019 Oct 17;11(10). pii: v11100960. doi: 10.3390/v11100960.
1999-4915
31627415
10.3390/v11100960
http://hdl.handle.net/10033/622004
Viruses
Yellow fever virus (YFV) represents a re-emerging zoonotic pathogen, transmitted by mosquito vectors to humans from primate reservoirs. Sporadic outbreaks of YFV occur in endemic tropical regions, causing a viral hemorrhagic fever (VHF) associated with high mortality rates. Despite a highly effective vaccine, no antiviral treatments currently exist. Therefore, YFV represents a neglected tropical disease and is chronically understudied, with many aspects of YFV biology incompletely defined including host range, host-virus interactions and correlates of host immunity and pathogenicity. In this article, we review the current state of YFV research, focusing on the viral lifecycle, host responses to infection, species tropism and the success and associated limitations of the YFV-17D vaccine. In addition, we highlight the current lack of available treatments and use publicly available sequence and structural data to assess global patterns of YFV sequence diversity and identify potential drug targets. Finally, we discuss how technological advances, including real-time epidemiological monitoring of outbreaks using next-generation sequencing and CRISPR/Cas9 modification of vector species, could be utilized in future battles against this re-emerging pathogen which continues to cause devastating disease
en
MPDI
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
E protein structure
animal models
flavivirus entry factor
global diversity
host immune response
re-emerging virus
transmission and vector control
vaccine
yellow fever virus
yellow fever virus tropism
Yellow Fever: Integrating Current Knowledge with Technological Innovations to Identify Strategies for Controlling a Re-Emerging Virus.
Article
Viruses
oai:repository.helmholtz-hzi.de:10033/6220232021-07-29T13:22:22Zcom_10033_620589com_10033_620857col_10033_622968col_10033_620590
Wibowo, Joko T
Kellermann, Matthias Y
Versluis, Dennis
Putra, Masteria Y
Murniasih, Tutik
Mohr, Kathrin I
Wink, Joachim
Engelmann, Michael
Praditya, Dimas F
Steinmann, Eike
Schupp, Peter J
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
2019-11-21T09:44:29Z
2019-11-21T09:44:29Z
2019-11-08
Mar Drugs. 2019 Nov 8;17(11). pii: md17110635. doi: 10.3390/md17110635.
1660-3397
31717405
10.3390/md17110635
http://hdl.handle.net/10033/622023
Marine Drugs
In order to minimize re-discovery of already known anti-infective compounds, we focused our screening approach on understudied, almost untapped marine environments including marine invertebrates and their associated bacteria. Therefore, two sea cucumber species, Holothuria leucospilota and Stichopus vastus, were collected from Lampung (Indonesia), and 127 bacterial strains were identified by partial 16S rRNA-gene sequencing analysis and compared with the NCBI database. In addition, the overall bacterial diversity from tissue samples of the sea cucumbers H. leucospilota and S. vastus was analyzed using the cultivation-independent Illumina MiSEQ analysis. Selected bacterial isolates were grown to high densities and the extracted biomass was tested against a selection of bacteria and fungi as well as the hepatitis C virus (HCV). Identification of putative bioactive bacterial-derived compounds were performed by analyzing the accurate mass of the precursor/parent ions (MS1) as well as product/daughter ions (MS2) using high resolution mass spectrometry (HRMS) analysis of all active fractions. With this attempt we were able to identify 23 putatively known and two previously unidentified precursor ions. Moreover, through 16S rRNA-gene sequencing we were able to identify putatively novel bacterial species from the phyla Actinobacteria, Proteobacteria and also Firmicutes. Our findings suggest that sea cucumbers like H. leucospilota and S. vastus are promising sources for the isolation of novel bacterial species that produce compounds with potentially high biotechnological potential.
en
MPDI
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
anti-infective marine derived compounds
de-replication
marine bacteria
mass spectrometry
sea cucumber
Biotechnological Potential of Bacteria Isolated from the Sea Cucumber and from Lampung, Indonesia.
Article
Marine drugs2019-11-21T09:44:30Zoai:repository.helmholtz-hzi.de:10033/6220282019-11-26T02:09:24Zcom_10033_620533com_10033_620589com_10033_620618col_10033_620534col_10033_620622col_10033_620608col_10033_620590
Prochnow, Hans
Rox, Katharina
Birudukota, N V Suryanarayana
Weichert, Loreen
Hotop, Sven-Kevin
Klahn, Philipp
Mohr, Kathrin
Franz, Sergej
Banda, Dominic H
Blockus, Sebastian
Schreiber, Janine
Haid, Sibylle
Oeyen, Merel
Martinez, Javier P
Süssmuth, Roderich D
Wink, Joachim
Meyerhans, Andreas
Goffinet, Christine
Messerle, Martin
Schulz, Thomas F
Kröger, Andrea
Schols, Dominique
Pietschmann, Thomas
Brönstrup, Mark
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
2019-11-25T13:38:21Z
2019-11-25T13:38:21Z
2019-10-30
J Virol. 2019 Oct 30. pii: JVI.01471-19. doi: 10.1128/JVI.01471-19.
1098-5514
31666384
10.1128/JVI.01471-19
http://hdl.handle.net/10033/622028
Journal of Virology
To counteract the serious health threat posed by known and novel viral pathogens, drugs that target a variety of viruses through a common mechanism have attracted recent attention due to their potential in treating (re-)emerging infections, for which direct acting antivirals are not available. We found that labyrinthopeptins A1 and A2, the prototype congeners of carbacyclic lanthipeptides, inhibit the proliferation of diverse enveloped viruses, including Dengue virus, Zika virus, West Nile virus, Hepatitis C virus, Chikungunya virus, Karposi's Sarcoma-associated Herpes virus, Cytomegalovirus, and Herpes Simplex virus, in the low μM to nM range. Mechanistic studies on viral particles revealed that labyrinthopeptins induce a virolytic effect through binding to the viral membrane lipid phosphatidylethanolamine (PE). These effects are enhanced by a combined equimolar application of both labyrinthopeptins, and a clear synergism was observed across a concentration range corresponding to IC10-IC90 values of the compounds. Time-resolved experiments with large unilamellar vesicles (LUVs) reveal that membrane lipid raft compositions (PC/PE/Chol/SM (17:10:33:40)) are particularly sensitive to labyrinthopeptins compared to PC/PE (90:10) LUVs, even though the overall PE-amount remains constant. Labyrinthopeptins exhibited low cytotoxicity and had favorable pharmacokinetic properties in mice (t1/2= 10.0 h), which designates them as promising antiviral compounds acting by an unusual viral lipid targeting mechanism.Importance For many viral infections, current treatment options are insufficient. Because the development of each antiviral drug is time-consuming and expensive, the prospect of finding broad-spectrum antivirals that can fight multiple, diverse viruses - well-known as well as (re-)emerging species - has gained attention, especially for the treatment of viral co-infections. While most known broad spectrum agents address processes in the host cell, we found that targeting lipids of the free virus outside the host cell with the natural products labyrinthopeptin A1 and A2 is a viable strategy to inhibit the proliferation of a broad range of viruses from different families, including Chikungunya virus, Dengue virus, Zika virus, Karposi's Sarcoma-associated Herpes virus, or Cytomegalovirus. Labyrinthopeptins bind to viral phosphatidylethanolamine and induce virolysis without exerting cytotoxicity to host cells. This represents a novel and unusual mechanism to tackle medically relevant viral infections.
en
ASM
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Labyrinthopeptins exert broad-spectrum antiviral activity through lipid-binding-mediated virolysis.
Article
Journal of virology
oai:repository.helmholtz-hzi.de:10033/6220342019-11-28T01:59:56Zcom_10033_620589col_10033_620590
Badenhorst, Marcha
de Heus, Phebe
Auer, Angelika
Rümenapf, Till
Tegtmeyer, Birthe
Kolodziejek, Jolanta
Nowotny, Norbert
Steinmann, Eike
Cavalleri, Jessika M.V.
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Badenhorst, M.
2019-11-27T09:51:17Z
2019-11-27T09:51:17Z
2019-11-01
Viruses. 2019 Nov 1;11(11). pii: v11111014. doi: 10.3390/v11111014.
31683893
10.3390/v11111014
https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85074548047&origin=inward
http://hdl.handle.net/10033/622034
Viruses
2-s2.0-85074548047
SCOPUS_ID:85074548047
Prevalence studies have demonstrated a global distribution of equine hepacivirus (EqHV), a member of the family Flaviviridae. However, apart from a single case of vertical transmission, natural routes of EqHV transmission remain elusive. Many known flaviviruses are horizontally transmitted between hematophagous arthropods and vertebrate hosts. This study represents the first investigation of potential EqHV transmission by mosquitoes. More than 5000 mosquitoes were collected across Austria and analyzed for EqHV ribonucleic acid (RNA) by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Concurrently, 386 serum samples from horses in eastern Austria were analyzed for EqHV-specific antibodies by luciferase immunoprecipitation system (LIPS) and for EqHV RNA by RT-qPCR. Additionally, liver-specific biochemistry parameters were compared between EqHV RNA-positive horses and EqHV RNA-negative horses. Phylogenetic analysis was conducted in comparison to previously published sequences from various origins. No EqHV RNA was detected in mosquito pools. Serum samples yielded an EqHV antibody prevalence of 45.9% (177/386) and RNA prevalence of 4.15% (16/386). EqHV RNA-positive horses had significantly higher glutamate dehydrogenase (GLDH) levels (p = 0.013) than control horses. Phylogenetic analysis showed high similarity between nucleotide sequences of EqHV in Austrian horses and EqHV circulating in other regions. Despite frequently detected evidence of EqHV infection in Austrian horses, no viral RNA was found in mosquitoes. It is therefore unlikely that mosquitoes are vectors of this flavivirus.
en
MPDI
Viruses
11
11
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
arbovirus
flavivirus
hematophagous arthropod
hepacivirus A
hepatitis
insects
mosquito-borne virus
virus transmission
No Evidence of Mosquito Involvement in the Transmission of Equine Hepacivirus (Flaviviridae) in an Epidemiological Survey of Austrian Horses
Article
oai:repository.helmholtz-hzi.de:10033/6220352019-11-28T02:00:06Zcom_10033_620589col_10033_620590
Gerold, Gisa
Moeller, Rebecca
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-11-27T10:29:34Z
2019-11-27T10:29:34Z
2019-08-19
Cold Spring Harb Perspect Med. 2019 Aug 19. pii: cshperspect.a036830. doi: 10.1101/cshperspect.a036830.
2157-1422
31427285
10.1101/cshperspect.a036830
http://hdl.handle.net/10033/622035
Cold Spring Harbor Perspectives in Medicine
Hepatitis C virus (HCV) entry is among the best-studied uptake processes for human pathogenic viruses. Uptake follows a spatially and temporally tightly controlled program. Numerous host factors including proteins, lipids, and glycans promote productive uptake of HCV particles into human liver cells. The virus initially attaches to surface proteoglycans, lipid receptors such as the scavenger receptor BI (SR-BI), and to the tetraspanin CD81. After lateral translocation of virions to tight junctions, claudin-1 (CLDN1) and occludin (OCLN) are essential for entry. Clathrin-mediated endocytosis engulfs HCV particles, which fuse with endosomal membranes after pH drop. Uncoating of the viral RNA genome in the cytoplasm completes the entry process. Here we systematically review and classify HCV entry factors by their mechanistic role, relevance, and level of evidence. Finally, we report on more recent knowledge on determinants of membrane fusion and close with an outlook on future implications of HCV entry research.
en
Cold Spring Harbor Laboratory Press
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Hepatitis C Virus Entry: Protein Interactions and Fusion Determinants Governing Productive Hepatocyte Invasion.
Article
Cold Spring Harbor perspectives in medicine
oai:repository.helmholtz-hzi.de:10033/6220842020-01-16T03:07:50Zcom_10033_620589col_10033_620590
Todt, Daniel
Friesland, Martina
Moeller, Nora
Praditya, Dimas
Kinast, Volker
Brüggemann, Yannick
Knegendorf, Leonard
Burkard, Thomas
Steinmann, Joerg
Burm, Rani
Verhoye, Lieven
Wahid, Avista
Meister, Toni Luise
Engelmann, Michael
Pfankuche, Vanessa M
Puff, Christina
Vondran, Florian W R
Baumgärtner, Wolfgang
Meuleman, Philip
Behrendt, Patrick
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2020-01-15T13:52:03Z
2020-01-15T13:52:03Z
2020-01-02
Proc Natl Acad Sci U S A. 2020 Jan 2. pii: 1912307117. doi: 10.1073/pnas.1912307117.
1091-6490
31896581
10.1073/pnas.1912307117
http://hdl.handle.net/10033/622084
Proceedings of the National Academy of sciences
Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and the leading cause for acute viral hepatitis worldwide. The virus is classified as a member of the genus Orthohepevirus A within the Hepeviridae family. Due to the absence of a robust cell culture model for HEV infection, the analysis of the viral life cycle, the development of effective antivirals and a vaccine is severely limited. In this study, we established a protocol based on the HEV genotype 3 p6 (Kernow C-1) and the human hepatoma cell lines HepG2 and HepG2/C3A with different media conditions to produce intracellular HEV cell culture-derived particles (HEVcc) with viral titers between 105 and 106 FFU/mL. Viral titers could be further enhanced by an HEV variant harboring a mutation in the RNA-dependent RNA polymerase. These HEVcc particles were characterized in density gradients and allowed the trans-complementation of subgenomic reporter HEV replicons. In addition, in vitro produced intracellular-derived particles were infectious in liver-humanized mice with high RNA copy numbers detectable in serum and feces. Efficient infection of primary human and swine hepatocytes using the developed protocol could be observed and was inhibited by ribavirin. Finally, RNA sequencing studies of HEV-infected primary human hepatocytes demonstrated a temporally structured transcriptional defense response. In conclusion, this robust cell culture model of HEV infection provides a powerful tool for studying viral-host interactions that should facilitate the discovery of antiviral drugs for this important zoonotic pathogen.
en
National Academy of Sciences
info:eu-repo/grantAgreement/EC/H2020/643476
openAccess
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
hepatitis E virus (HEV)
humanized mice
infection
primary hepatocytes
transcriptomics
Robust hepatitis E virus infection and transcriptional response in human hepatocytes.
Article
Proceedings of the National Academy of Sciences of the United States of America
oai:repository.helmholtz-hzi.de:10033/6221102020-01-31T02:17:26Zcom_10033_620589col_10033_620590
Knegendorf, Leonard
Drave, Svenja A
Dao Thi, Viet Loan
Debing, Yannick
Brown, Richard J P
Vondran, Florian W R
Resner, Kathrin
Friesland, Martina
Khera, Tanvi
Engelmann, Michael
Bremer, Birgit
Wedemeyer, Heiner
Behrendt, Patrick
Neyts, Johan
Pietschmann, Thomas
Todt, Daniel
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2020-01-30T12:32:45Z
2020-01-30T12:32:45Z
2018-01-01
Hepatol Commun. 2018 Jan 8;2(2):173-187. doi: 10.1002/hep4.1138. eCollection 2018 Feb.
2471-254X
29404525
10.1002/hep4.1138
http://hdl.handle.net/10033/622110
Hepatology Communications
Hepatitis E virus (HEV) is a member of the genus Orthohepevirus in the family Hepeviridae and the causative agent of hepatitis E in humans. HEV is a major health problem in developing countries, causing mortality rates up to 25% in pregnant women. However, these cases are mainly reported for HEV genotype (gt)1, while gt3 infections are usually associated with subclinical courses of disease. The pathogenic mechanisms of adverse maternal and fetal outcome during pregnancy in HEV-infected pregnant women remain elusive. In this study, we observed that HEV is capable of completing the full viral life cycle in placental-derived cells (JEG-3). Following transfection of JEG-3 cells, HEV replication of both HEV gts could be observed. Furthermore, determination of extracellular and intracellular viral capsid levels, infectivity, and biophysical properties revealed production of HEV infectious particles with similar characteristics as in liver-derived cells. Viral entry was analyzed by infection of target cells and detection of either viral RNA or staining for viral capsid protein by immunofluorescence. HEV gt1 and gt3 were efficiently inhibited by ribavirin in placental as well as in human hepatoma cells. In contrast, interferon-α sensitivity was lower in the placental cells compared to liver cells for gt1 but not gt3 HEV. Simultaneous determination of interferon-stimulated gene expression levels demonstrated an efficient HEV-dependent restriction in JEG-3. Conclusion: We showed differential tissue-specific host responses to HEV genotypes, adding to our understanding of the mechanisms contributing to fatal outcomes of HEV infections during pregnancy. Using this cell-culture system, new therapeutic options for HEV during pregnancy can be identified and evaluated. (Hepatology Communications 2018;2:173-187).
en
Wiley-Blackwell
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Hepatitis E virus replication and interferon responses in human placental cells.
Article
Hepatology communications
oai:repository.helmholtz-hzi.de:10033/6222082020-03-18T02:01:52Zcom_10033_620589col_10033_620590
Neumann, Ariane
Brogden, Graham
von Köckritz-Blickwede, Maren
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2020-03-17T13:44:17Z
2020-03-17T13:44:17Z
2020-02-18
Biology (Basel). 2020 Feb 18;9(2). pii: biology9020034. doi: 10.3390/biology9020034.
2079-7737
32085405
10.3390/biology9020034
http://hdl.handle.net/10033/622208
Biology
The discovery, in 2004, of extracellular traps released by neutrophils has extended our understanding of the mode of action of various innate immune cells. This fascinating discovery demonstrated the extracellular trapping and killing of various pathogens by neutrophils. During the last decade, evidence has accumulated showing that extracellular traps play a crucial role in the defence mechanisms of various cell types present in vertebrates, invertebrates, and plants. The aim of this review is to summarise the relevant literature on the evolutionary history of extracellular traps used as a weapon in various kingdoms of life.
en
MDPI
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
conserved mechanisms
defence mechanism
innate immunity
neutrophil extracellular traps (NETs)
tree of life
Extracellular Traps: An Ancient Weapon of Multiple Kingdoms.
Article
Other
9
2
Biology
Switzerland
oai:repository.helmholtz-hzi.de:10033/6222312020-04-16T02:15:39Zcom_10033_620589col_10033_620590
Carpentier, Arnaud
Sheldon, Julie
Vondran, Florian W R
Brown, Richard Jp
Pietschmann, Thomas
TWINCORE, Zentrum für Experimentelle und Klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2020-04-15T11:07:28Z
2020-04-15T11:07:28Z
2020-02-29
Gut. 2020 Feb 29. pii: gutjnl-2019-319354. doi: 10.1136/gutjnl-2019-319354.
32114504
10.1136/gutjnl-2019-319354
http://hdl.handle.net/10033/622231
1468-3288
Gut
Transcriptional profiling revealed that HLCs constitutively express messenger RNA of RLRs, and members of the IFN pathway. Moreover, HLCs upregulated IFNs and canonical interferon-regulated genes (IRGs) upon transfection with the double-stranded RNA mimic poly(I:C). Infection of HLCs with Jc1-HCVcc produced only limited viral progeny. In contrast, infection with p100, a Jc1-derived virus population with enhanced replication fitness and partial resistance to IFN, resulted in robust yet transient viraemia. Viral titres declined concomitant with a peak of IRG induction. Addition of ruxolitinib, a JAK/STAT inhibitor, permitted chronic infection and raised p100 infectious virus titres to 1×105 FFU/mL. IRGs expression profiling in infected HLCs revealed a landscape of HCV-dependent transcriptional changes similar to HCV-infected primary human hepatocytes, but distinct from Huh-7.5 cells. Withdrawal of ruxolitinib restored innate immune responses and resulted in HCV clearance.
en
BMJ Publishing Group
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
HCV
hepatocyte
immune response
interferon
stem cells
Efficient acute and chronic infection of stem cell-derived hepatocytes by hepatitis C virus.
Article
Gut
England
oai:repository.helmholtz-hzi.de:10033/6222732020-05-26T01:29:33Zcom_10033_620533com_10033_620857com_10033_620626com_10033_620589col_10033_620534col_10033_620858col_10033_620629col_10033_620590
Blockus, Sebastian
Sake, Svenja M
Wetzke, Martin
Grethe, Christina
Graalmann, Theresa
Pils, Marina
Le Goffic, Ronan
Galloux, Marie
Prochnow, Hans
Rox, Katharina
Hüttel, Stephan
Rupcic, Zeljka
Wiegmann, Bettina
Dijkman, Ronald
Rameix-Welti, Marie-Anne
Eléouët, Jean-François
Duprex, W Paul
Thiel, Volker
Hansen, Gesine
Brönstrup, Mark
Haid, Sibylle
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2020-05-25T14:26:43Z
2020-05-25T14:26:43Z
2020-03-18
Antiviral Res. 2020;177:104774. doi:10.1016/j.antiviral.2020.104774.
32197980
10.1016/j.antiviral.2020.104774
http://hdl.handle.net/10033/622273
1872-9096
Antiviral research
Acute lower respiratory tract infections (ALRI) caused by respiratory syncytial virus (RSV) are associated with a severe disease burden among infants and elderly patients. Treatment options are limited. While numerous drug candidates with different viral targets are under development, the utility of RSV entry inhibitors is challenged by a low resistance barrier and by single mutations causing cross-resistance against a wide spectrum of fusion inhibitor chemotypes. We developed a cell-based screening assay for discovery of compounds inhibiting infection with primary RSV isolates. Using this system, we identified labyrinthopeptin A1 and A2 (Laby A1/A2), lantibiotics isolated from Actinomadura namibiensis, as effective RSV cell entry inhibitors with IC50s of 0.39 μM and 4.97 μM, respectively, and with favourable therapeutic index (>200 and > 20, respectively). Both molecules were active against multiple RSV strains including primary isolates and their antiviral activity against RSV was confirmed in primary human airway cells ex vivo and a murine model in vivo. Laby A1/A2 were antiviral in prophylactic and therapeutic treatment regimens and displayed synergistic activity when applied in combination with each other. Mechanistic studies showed that Laby A1/A2 exert virolytic activity likely by binding to phosphatidylethanolamine moieties within the viral membrane and by disrupting virus particle membrane integrity. Probably due to its specific mode of action, Laby A1/A2 antiviral activity was not affected by common resistance mutations to known RSV entry inhibitors. Taken together, Laby A1/A2 represent promising candidates for development as RSV inhibitors. Moreover, the cell-based screening system with primary RSV isolates described here should be useful to identify further antiviral agents.
en
Elsevier
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Antiviral activity
Human respiratory syncytial virus (hRSV)
Labyrinthopeptin
Lanthipeptide
Virolytic
Virus entry
Labyrinthopeptins as virolytic inhibitors of respiratory syncytial virus cell entry.
Article
177
104774
Antiviral research
Netherlands
oai:repository.helmholtz-hzi.de:10033/6223172020-07-01T02:33:47Zcom_10033_620533com_10033_620589col_10033_620534col_10033_620590
Gunesch, Antonia P
Zapatero-Belinchon, Francisco J
Pinkert, Lukas
Steinmann, Eike
Manns, Michael P
Schneider, Gisbert
Pietschmann, Thomas
Brönstrup, Mark
von Hahn, Thomas
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.; TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2020-06-29T09:16:05Z
2020-06-29T09:16:05Z
2020-06-08
Antimicrob Agents Chemother. 2020;AAC.00143-20. doi:10.1128/AAC.00143-20.
32513799
http://hdl.handle.net/10033/622317
1098-6596
Antimicrobial agents and chemotherapy
Several cationic amphiphilic drugs (CADs) have been found to inhibit cell entry of filoviruses and other enveloped viruses. Structurally unrelated CADs may have antiviral activity, yet the underlying common mechanism and structure-activity relationship are incompletely understood.We aimed to understand how widespread antiviral activity is among CADs and which structural and physico-chemical properties are linked to entry inhibition.We measured inhibition of Marburg virus pseudoparticle (MARVpp) cell entry by 45 heterogeneous and mostly FDA-approved CADs and cytotoxicity in EA.hy926 cells. We analysed correlation of antiviral activity with four chemical properties: pKa, ClogP, molecular weight and distance between the basic group and hydrophobic ring structures. Additionally, we quantified drug-induced phospholipidosis (DIPL) of a CAD subset by flow cytometry. Structurally similar compounds (derivatives) and those with similar chemical properties but unrelated structure (analogues) to strong inhibitors were obtained by two in silico similarity search approaches and tested for antiviral activity. Overall 11 out of 45 (24 %) CADs inhibited MARVpp by 40 % or more. The strongest antiviral compounds were dronedarone, triparanol and quinacrine. Structure-activity relationship studies revealed highly significant correlations between antiviral activity, hydrophobicity (ClogP>4), and DIPL. Moreover, pKa and intra-molecular distance between hydrophobic and hydrophilic moieties correlated with antiviral activity, but to a lesser extent. We also showed that in contrast to analogues, derivatives had similar antiviral activity as the seed compound dronedarone. Overall, one quarter of CADs inhibits MARVpp entry in vitro and antiviral activity of CADs mostly relies on their hydrophobicity, yet is promoted by the individual structure.
en
ASM
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Filovirus antiviral activity of cationic amphiphilic drugs is associated with lipophilicity and ability to induce phospholipidosis.
Article
Antimicrobial agents and chemotherapy
United States
oai:repository.helmholtz-hzi.de:10033/6223472020-07-11T02:50:49Zcom_10033_620589col_10033_620590
Bartenschlager, Ralf
Baumert, Thomas F
Bukh, Jens
Houghton, Michael
Lemon, Stanley M
Lindenbach, Brett D
Lohmann, Volker
Moradpour, Darius
Pietschmann, Thomas
Rice, Charles M
Thimme, Robert
Wakita, Takaji
TwinCore, Zentrum für experimentelle und klinische Infektionsforchung GmbH, Feodor-Lynen-Str.7, 30625 Hannover, Germany.
2020-07-10T09:38:19Z
2020-07-10T09:38:19Z
2018-03-02
Virus Res. 2018;248:53-62. doi:10.1016/j.virusres.2018.02.016.
29477639
10.1016/j.virusres.2018.02.016
http://hdl.handle.net/10033/622347
1872-7492
Virus research
The development and clinical implementation of direct-acting antivirals (DAAs) has revolutionized the treatment of chronic hepatitis C. Infection with any hepatitis C virus (HCV) genotype can now be eliminated in more than 95% of patients with short courses of all-oral, well-tolerated drugs, even in those with advanced liver disease and liver transplant recipients. DAAs have proven so successful that some now consider HCV amenable to eradication, and continued research on the virus of little remaining medical relevance. However, given 400,000 HCV-related deaths annually important challenges remain, including identifying those who are infected, providing access to treatment and reducing its costs. Moreover, HCV infection rarely induces sterilizing immunity, and those who have been cured with DAAs remain at risk for reinfection. Thus, it is very unlikely that global eradication and elimination of the cancer risk associated with HCV infection can be achieved without a vaccine, yet research in that direction receives little attention. Further, over the past two decades HCV research has spearheaded numerous fundamental discoveries in the fields of molecular and cell biology, immunology and microbiology. It will continue to do so, given the unique opportunities afforded by the reagents and knowledge base that have been generated in the development and clinical application of DAAs. Considering these critical challenges and new opportunities, we conclude that funding for HCV research must be sustained.
en
Elsevier
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Direct acting antiviral therapy
HCV research funding
HCV vaccine
Immune reconstitution
Critical challenges and emerging opportunities in hepatitis C virus research in an era of potent antiviral therapy: Considerations for scientists and funding agencies.
Article
Other
248
53
62
Virus research
Netherlands
oai:repository.helmholtz-hzi.de:10033/6223542020-07-22T02:29:43Zcom_10033_620589col_10033_620590
Haid, Sibylle
Grethe, Christina
Bankwitz, Dorothea
Grunwald, Thomas
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2020-07-21T09:34:39Z
2020-07-21T09:34:39Z
2015-12-30
J Virol. 2015;90(6):3065-3073. Published 2015 Dec 30. doi:10.1128/JVI.03074-15.
26719246
10.1128/JVI.03074-15
http://hdl.handle.net/10033/622354
1098-5514
Journal of virology
Lentiviral budding is governed by group-specific antigens (Gag proteins) and proceeds in the absence of cognate viral envelope proteins, which has been exploited to create pseudotypes incorporating envelope proteins from nonlentiviral families. Here, we report the generation of infectious lentiviral pseudoparticles incorporating human respiratory syncytial virus (hRSV) F protein alone (hRSV-Fpp) or carrying SH, G, and F proteins (hRSV-SH/G/Fpp). These particles recapitulate key infection steps of authentic hRSV particles, including utilization of glycosaminoglycans and low-pH-independent cell entry. Moreover, hRSV pseudoparticles (hRSVpp) can faithfully reproduce phenotypic resistance to a small-molecule fusion inhibitor in clinical development (BMS-433771) and a licensed therapeutic F protein-targeting antibody (palivizumab). Inoculation of several human cell lines from lung and liver revealed more than 30-fold differences in susceptibility to hRSVpp infection, suggesting differential expression of hRSV entry cofactors and/or restriction factors between these cell types. Moreover, we observed cell-type-dependent functional differences between hRSVpp carrying solely F protein or SH, G, and F proteins with regard to utilization of glycosaminoglycans. Using hRSVpp, we identified penta-O-galloyl-β-d-glucose (PGG) as a novel hRSV cell entry inhibitor. Moreover, we show that PGG also inhibits cell entry of hRSVpp carrying F proteins resistant to BMS-433771 or palivizumab. This work sheds new light on the mechanisms of hRSV cell entry, including possible strategies for antiviral intervention. Moreover, hRSVpp should prove valuable to dissect hRSV envelope protein functions, including the interaction with cell entry factors.
Importance:
Lentiviral pseudotypes are highly useful to specifically dissect the functions of viral and host factors in cell entry, which have been exploited for numerous viruses. Here, we successfully created hRSVpp and show that they faithfully recapitulate key characteristics of parental hRSV cell entry. Importantly, hRSVpp accurately mirror hRSV resistance to small-molecule fusion inhibitors and clinically approved therapeutic antibodies. Moreover, we observed highly different susceptibilities of cell lines to hRSVpp infection and also differences between hRSVpp types (with F protein alone or with SH, G, and F proteins) in regard to cell entry. This indicates differential expression of host factors determining hRSV cell entry between these cell lines and highlights the fact that the hRSVpp system is useful to explore the functional properties of hRSV envelope protein combinations. Therefore, this system will be highly useful to study hRSV cell entry and host factor usage and to explore antiviral strategies targeting hRSV cell entry.
en
ASM
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Identification of a Human Respiratory Syncytial Virus Cell Entry Inhibitor by Using a Novel Lentiviral Pseudotype System.
Article
Other
90
6
3065
73
Journal of virology
United States
oai:repository.helmholtz-hzi.de:10033/6223622020-07-25T01:29:34Zcom_10033_620589col_10033_620590
Vieyres, Gabrielle
Reichert, Isabelle
Carpentier, Arnaud
Vondran, Florian W R
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2020-07-24T13:22:48Z
2020-07-24T13:22:48Z
2020-06-15
PLoS Pathog. 2020;16(6):e1008554. Published 2020 Jun 15. doi:10.1371/journal.ppat.1008554.
32542055
10.1371/journal.ppat.1008554
http://hdl.handle.net/10033/622362
1553-7374
PLoS pathogens
Lipid droplets are essential cellular organelles for storage of fatty acids and triglycerides. The hepatitis C virus (HCV) translocates several of its proteins onto their surface and uses them for production of infectious progeny. We recently reported that the lipid droplet-associated α/β hydrolase domain-containing protein 5 (ABHD5/CGI-58) participates in HCV assembly by mobilizing lipid droplet-associated lipids. However, ABHD5 itself has no lipase activity and it remained unclear how ABHD5 mediates lipolysis critical for HCV assembly. Here, we identify adipose triglyceride lipase (ATGL) as ABHD5 effector and new host factor involved in the hepatic lipid droplet degradation as well as in HCV and lipoprotein morphogenesis. Modulation of ATGL protein expression and lipase activity controlled lipid droplet lipolysis and virus production. ABHD4 is a paralog of ABHD5 unable to activate ATGL or support HCV assembly and lipid droplet lipolysis. Grafting ABHD5 residues critical for activation of ATGL onto ABHD4 restored the interaction between lipase and co-lipase and bestowed the pro-viral and lipolytic functions onto the engineered protein. Congruently, mutation of the predicted ABHD5 protein interface to ATGL ablated ABHD5 functions in lipid droplet lipolysis and HCV assembly. Interestingly, minor alleles of ABHD5 and ATGL associated with neutral lipid storage diseases in human, are also impaired in lipid droplet lipolysis and their pro-viral functions. Collectively, these results show that ABHD5 cooperates with ATGL to mobilize triglycerides for HCV infectious virus production. Moreover, viral manipulation of lipid droplet homeostasis via the ABHD5-ATGL axis, akin to natural genetic variation in these proteins, emerges as a possible mechanism by which chronic HCV infection causes liver steatosis.
en
PLOS
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
The ATGL lipase cooperates with ABHD5 to mobilize lipids for hepatitis C virus assembly.
Article
16
6
e1008554
PLoS pathogens
United States
oai:repository.helmholtz-hzi.de:10033/6223692020-07-30T02:31:41Zcom_10033_620589col_10033_620590
Armando, Federico
Gambini, Matteo
Corradi, Attilio
Becker, Kathrin
Marek, Katarzyna
Pfankuche, Vanessa Maria
Mergani, Ahmed Elmonastir
Brogden, Graham
de Buhr, Nicole
Von Köckritz-Blickwede, Maren
Naim, Hassan Y
Baumgärtner, Wolfgang
Puff, Christina
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2020-07-29T09:36:45Z
2020-07-29T09:36:45Z
2020-07-06
J Cell Mol Med. 2020;10.1111/jcmm.15585. doi:10.1111/jcmm.15585.
32627957
10.1111/jcmm.15585
http://hdl.handle.net/10033/622369
1582-4934
Journal of cellular and molecular medicine
Sarcomas especially of histiocytic origin often possess a poor prognosis and response to conventional therapies. Interestingly, tumours undergoing mesenchymal to epithelial transition (MET) are often associated with a favourable clinical outcome. This process is characterized by an increased expression of epithelial markers leading to a decreased invasion and metastatic rate. Based on the failure of conventional therapies, viral oncolysis might represent a promising alternative with canine distemper virus (CDV) as a possible candidate. This study hypothesizes that a CDV infection of canine histiocytic sarcoma cells (DH82 cells) triggers the MET process leading to a decreased cellular motility. Immunofluorescence and immunoblotting were used to investigate the expression of epithelial and mesenchymal markers followed by scratch assay and an invasion assay as functional confirmation. Furthermore, microarray data were analysed for genes associated with the MET process, invasion and angiogenesis. CDV-infected cells exhibited an increased expression of epithelial markers such as E-cadherin and cytokeratin 8 compared to controls, indicating a MET process. This was accompanied by a reduced cell motility and invasiveness. Summarized, these results suggest that CDV infection of DH82 cells triggers the MET process by an increased expression of epithelial markers resulting in a decreased cell motility in vitro.
en
Blackwell Publishing
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
3D modelling
DH82
canine distemper virus
canine histiocytic sarcoma
mesenchymal to epithelial transition
viral oncolysis
Mesenchymal to epithelial transition driven by canine distemper virus infection of canine histiocytic sarcoma cells contributes to a reduced cell motility in vitro.
Article
Journal of cellular and molecular medicine
England
dim///col_10033_620590/100