2024-03-28T23:03:14Zhttp://repository.helmholtz-hzi.de/oai/requestoai:repository.helmholtz-hzi.de:10033/784942019-08-30T11:30:58Zcom_10033_620589col_10033_620590
Low pH-dependent hepatitis C virus membrane fusion depends on E2 integrity, target lipid composition, and density of virus particles.
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.
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
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.
2009-08-25T13:22:55Z
2009-08-25T13:22:55Z
2009-06-26
Article
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
en
oai:repository.helmholtz-hzi.de:10033/1076192019-08-30T11:30:32Zcom_10033_620589col_10033_620590
Glucocorticosteroids increase cell entry by hepatitis C virus.
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.
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
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.
2010-07-14T09:43:57Z
2010-07-14T09:43:57Z
2010-05
Article
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
en
oai:repository.helmholtz-hzi.de:10033/1181942019-08-30T11:30:58Zcom_10033_620589col_10033_620590
How stable is the hepatitis C virus (HCV)? Environmental stability of HCV and its susceptibility to chemical biocides.
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.
Cell Line
Disinfectants
Hepacivirus
Humans
Microbial Viability
RNA, Viral
Reverse Transcriptase Polymerase Chain Reaction
Temperature
Time Factors
Transfection
Virus Inactivation
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.
2010-12-21T15:25:11Z
2010-12-21T15:25:11Z
2010-06-15
Article
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
en
oai:repository.helmholtz-hzi.de:10033/1463552019-08-30T11:27:46Zcom_10033_620589col_10033_620590
Completion of hepatitis C virus replication cycle in heterokaryons excludes dominant restrictions in human non-liver and mouse liver cell lines.
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.
Animals
Cell Fusion
Cell Line
HEK293 Cells
Hela Cells
Hepacivirus
Humans
Interferon-alpha
Mice
Models, Animal
Transfection
Virus Replication
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.
2011-10-21T13:45:31Z
2011-10-21T13:45:31Z
2011-04
Article
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
en
oai:repository.helmholtz-hzi.de:10033/2136952019-08-30T11:37:23Zcom_10033_620589col_10033_620590
The novel immunosuppressive protein kinase C inhibitor sotrastaurin has no pro-viral effects on the replication cycle of hepatitis B or C virus.
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.
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 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.
2012-03-01T10:28:46Z
2012-03-01T10:28:46Z
2011
Article
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
en
oai:repository.helmholtz-hzi.de:10033/2139892019-08-30T11:30:58Zcom_10033_620589col_10033_620590
Outbreak due to a Klebsiella pneumoniae strain harbouring KPC-2 and VIM-1 in a German university hospital, July 2010 to January 2011.
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
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
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.
2012-03-02T10:50:33Z
2012-03-02T10:50:33Z
2011
Article
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
en
oai:repository.helmholtz-hzi.de:10033/2164742019-08-30T11:32:41Zcom_10033_620589col_10033_620590
Inactivation and survival of hepatitis C virus on inanimate surfaces.
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.
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
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.
2012-03-23T14:33:51Z
2012-03-23T14:33:51Z
2011-12-15
Article
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
en
Archived with thanks to The Journal of infectious diseases
oai:repository.helmholtz-hzi.de:10033/2229982019-08-30T11:24:31Zcom_10033_620589col_10033_620590
Bile Acids Specifically Increase Hepatitis C Virus RNA-Replication.
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.
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.
2012-05-10T09:24:05Z
2012-05-10T09:24:05Z
2012
Article
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
en
Archived with thanks to PloS one
oai:repository.helmholtz-hzi.de:10033/2256022019-08-30T11:27:46Zcom_10033_620589col_10033_620590
A Plant-Derived Flavonoid Inhibits Entry of All HCV Genotypes Into Human Hepatocytes.
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.
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.
2012-05-23T14:24:09Z
2012-05-23T14:24:09Z
2012-03-27
Article
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
ENG
Archived with thanks to Gastroenterology
oai:repository.helmholtz-hzi.de:10033/2470122019-08-30T11:24:31Zcom_10033_620589col_10033_620590
MAP-Kinase Regulated Cytosolic Phospholipase A2 Activity Is Essential for Production of Infectious Hepatitis C Virus Particles.
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.
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.
2012-10-04T14:40:26Z
2012-10-04T14:40:26Z
2012-07
Article
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
en
Archived with thanks to PLoS pathogens
oai:repository.helmholtz-hzi.de:10033/2655132019-08-30T11:24:31Zcom_10033_620589col_10033_620590
Specific acquisition of functional CD59 but not CD46 or CD55 by hepatitis C virus.
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.
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.
2013-01-15T15:33:08Z
2013-01-15T15:33:08Z
2012
Article
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
en
Archived with thanks to PloS one
oai:repository.helmholtz-hzi.de:10033/2703122019-08-30T11:25:11Zcom_10033_620589col_10033_620590
Mutations that alter use of hepatitis C virus cell entry factors mediate escape from neutralizing antibodies.
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.
Antibodies, Neutralizing
Cell Line, Tumor
Hepacivirus
Hepatitis C
Humans
Male
Mutation
Transplants
Virus Internalization
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.
2013-02-25T10:38:41Z
2013-02-25T10:38:41Z
2012-07
Article
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
en
Archived with thanks to Gastroenterology
oai:repository.helmholtz-hzi.de:10033/2705122019-08-30T11:33:29Zcom_10033_620589col_10033_620590
Subcellular localization and function of an epitope-tagged p7 viroporin in hepatitis C virus-producing cells.
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.
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.
2013-02-26T15:43:22Z
2013-02-26T15:43:22Z
2013-02
Article
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
en
Archived with thanks to Journal of virology
oai:repository.helmholtz-hzi.de:10033/2830332019-08-30T11:35:39Zcom_10033_620589col_10033_620590
Anti-infective properties of epigallocatechin-3-gallate (EGCG), a component of green tea.
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
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.
2013-04-18T13:14:48Z
2013-04-18T13:14:48Z
2013-03
Article
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
en
Archived with thanks to British journal of pharmacology
oai:repository.helmholtz-hzi.de:10033/2884822019-08-30T11:33:55Zcom_10033_620589col_10033_620590
Hepatitis C virus NS5B polymerase primes innate immune signaling.
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
Article
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.)
oai:repository.helmholtz-hzi.de:10033/2930052019-08-30T11:24:31Zcom_10033_620589col_10033_620590
Hepatitis C Virus p7 is Critical for Capsid Assembly and Envelopment.
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.
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.
2013-05-29T10:36:29Z
2013-05-29T10:36:29Z
2013-05
Article
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
en
Archived with thanks to PLoS pathogens
oai:repository.helmholtz-hzi.de:10033/2944392019-08-30T11:33:05Zcom_10033_620589col_10033_620590
The postbinding activity of scavenger receptor class B type I mediates initiation of hepatitis C virus infection and viral dissemination.
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.
Animals
Antibodies, Monoclonal
Antigens, CD36
Cell Line
Cholesterol, HDL
Hepacivirus
Hepatitis C
Humans
Lipoproteins, HDL
Mice
Rats
Receptors, Lipoprotein
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.
2013-06-24T09:46:00Z
2013-06-24T09:46:00Z
2013-02
Article
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.)
en
Archived with thanks to Hepatology (Baltimore, Md.)
oai:repository.helmholtz-hzi.de:10033/2947722019-08-30T11:33:05Zcom_10033_620589col_10033_620590
In vitro activity of colistin as single agent and in combination with antifungals against filamentous fungi occurring in patients with cystic fibrosis.
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.
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.
2013-06-27T12:29:21Z
2013-06-27T12:29:21Z
2013-05
Article
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
en
Archived with thanks to Mycoses
oai:repository.helmholtz-hzi.de:10033/2959742019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Cell culture systems for hepatitis C virus.
Steinmann, Eike
Pietschmann, Thomas
Helmholtz Centre for Infection Research, Hannover, Germany.
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.
2013-07-15T11:00:42Z
2013-07-15T11:00:42Z
2013
Book chapter
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
en
Archived with thanks to Current topics in microbiology and immunology
oai:repository.helmholtz-hzi.de:10033/2968372019-08-30T11:25:11Zcom_10033_620589col_10033_620590
Characterization of the inhibition of hepatitis C virus entry by in vitro-generated and patient-derived oxidized low-density lipoprotein.
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.
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
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.
2013-07-23T09:33:06Z
2013-07-23T09:33:06Z
2013-05
Article
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.)
en
Archived with thanks to Hepatology (Baltimore, Md.)
oai:repository.helmholtz-hzi.de:10033/2980672019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Thermostability of seven hepatitis C virus genotypes in vitro and in vivo.
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.
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.
2013-08-13T09:48:31Z
2013-08-13T09:48:31Z
2013-07
Article
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
en
Archived with thanks to Journal of viral hepatitis
oai:repository.helmholtz-hzi.de:10033/2990862019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Entry and replication of recombinant hepatitis C viruses in cell culture.
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.
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.
2013-08-19T12:21:26Z
2013-08-19T12:21:26Z
2013-02
Article
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.)
en
Archived with thanks to Methods (San Diego, Calif.)
oai:repository.helmholtz-hzi.de:10033/3054922019-08-30T11:27:16Zcom_10033_620589col_10033_620590
Mannan binding lectin-associated serine protease 1 is induced by hepatitis C virus infection and activates human hepatic stellate cells.
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.
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.
2013-11-18T14:13:32Z
2013-11-18T14:13:32Z
2013-11
Article
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
en
Archived with thanks to Clinical and experimental immunology
oai:repository.helmholtz-hzi.de:10033/3054932019-08-30T11:34:19Zcom_10033_620589col_10033_620590
Stability and transmission of hepatitis C virus in different anesthetic agents.
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
article
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
oai:repository.helmholtz-hzi.de:10033/3057032019-08-30T11:25:11Zcom_10033_620589col_10033_620590
An alpaca nanobody inhibits hepatitis C virus entry and cell-to-cell transmission.
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.
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.
2013-11-22T15:45:21Z
2013-11-22T15:45:21Z
2013-09
Article
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.)
en
Archived with thanks to Hepatology (Baltimore, Md.)
oai:repository.helmholtz-hzi.de:10033/3234302019-08-30T11:24:31Zcom_10033_620589col_10033_620590
Hepatitis C virus hypervariable region 1 variants presented on hepatitis B virus capsid-like particles induce cross-neutralizing antibodies.
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
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.
2014-07-18T13:24:25Z
2014-07-18T13:24:25Z
2014
Article
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
en
Archived with thanks to PloS one
oai:repository.helmholtz-hzi.de:10033/3254832019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Molecular basis of interferon resistance in hepatitis C virus.
Perales, Celia
Beach, Nathan M
Sheldon, Julie
Domingo, Esteban
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.
2014-08-27T12:51:47Z
2014-08-27T12:51:47Z
2014-06-23
Article
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
ENG
Archived with thanks to Current opinion in virology
oai:repository.helmholtz-hzi.de:10033/3258982019-08-30T11:24:24Zcom_10033_620589col_10033_620590
The HCV Life Cycle: In vitro Tissue Culture Systems and Therapeutic Targets
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
Article
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
http://www.karger.com?doi=10.1159/000360830
Archived with thanks to Digestive Diseases
Karger
oai:repository.helmholtz-hzi.de:10033/3261732019-08-30T11:34:20Zcom_10033_620589col_10033_620590
Development of a high-throughput pyrosequencing assay for monitoring temporal evolution and resistance associated variant emergence in the Hepatitis C virus protease coding-region
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
Article
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
oai:repository.helmholtz-hzi.de:10033/3328652019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Adaptation of Stenotrophomonas maltophilia in cystic fibrosis: molecular diversity, mutation frequency and antibiotic resistance.
Vidigal, P G
Dittmer, S
Steinmann, E
Buer, J
Rath, P-M
Steinmann, J
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.
2014-10-17T13:07:04Z
2014-10-17T13:07:04Z
2014-07
Article
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
en
Archived with thanks to International journal of medical microbiology : IJMM
oai:repository.helmholtz-hzi.de:10033/3330302019-08-30T11:33:30Zcom_10033_620589col_10033_620590
Value of multiplex PCR using cerebrospinal fluid for the diagnosis of ventriculostomy-related meningitis in neurosurgery patients.
Rath, P-M
Schoch, B
Adamzik, M
Steinmann, E
Buer, J
Steinmann, J
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).
2014-10-21T14:16:32Z
2014-10-21T14:16:32Z
2014-08
Article
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
en
Archived with thanks to Infection
oai:repository.helmholtz-hzi.de:10033/3370492019-08-30T11:27:16Zcom_10033_620589col_10033_620590
Hepatitis E in Germany--an under-reported infectious disease.
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.
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.
2014-12-11T09:23:59Z
2014-12-11T09:23:59Z
2014-09-01
Article
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
en
oai:repository.helmholtz-hzi.de:10033/3454092019-08-30T11:27:16Zcom_10033_620589col_10033_620590
The green tea polyphenol, epigallocatechin-3-gallate, inhibits hepatitis C virus entry.
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
Catechin
Cell Line, Tumor
Cells, Cultured
Hepacivirus
Hepatitis C
Humans
Tea
Virus Attachment
Virus Internalization
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.
2015-02-26T12:17:23Z
2015-02-26T12:17:23Z
2011-12
Article
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.)
en
oai:repository.helmholtz-hzi.de:10033/3464842019-08-30T11:33:30Zcom_10033_620589col_10033_620596
Mechanisms of methods for hepatitis C virus inactivation.
Pfaender, Stephanie
Brinkmann, Janine
Todt, Daniel
Riebesehl, Nina
Steinmann, Joerg
Steinmann, Jochen
Pietschmann, Thomas
Steinmann, Eike
Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research,Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
Virus inactivation by chemical disinfectants is an important instrument for infection control in medical settings, but the mechanisms involved are poorly understood. In this study, we systematically investigated the effects of several antiviral treatments on hepatitis C virus (HCV) particles as model for enveloped viruses. Studies were performed with authentic cell culture-derived viruses, and the influence of chemical disinfectants, heat, and UV treatment on HCV was analyzed by the determination of infectious particles in a limiting-dilution assay, by quantitative reverse transcription-PCR, by core enzyme-linked immunosorbent assay, and by proteolytic protection assay. All different inactivation methods resulted in a loss of HCV infectivity by targeting different parts of the virus particle. Alcohols such as ethanol and 2-propanol did not affect the viral RNA genome integrity but disrupted the viral envelope membrane in a capsid protection assay. Heat and UV treatment of HCV particles resulted in direct damage of the viral genome since transfection of viral particle-associated RNA into permissive cells did not initiate RNA replication. In addition, heat incubation at 80°C disrupted the HCV envelope, rendering the viral capsid susceptible to proteolytic digest. This study demonstrated the molecular processes of viral inactivation of an enveloped virus and should facilitate the development of effective disinfection strategies in infection control not only against HCV but also against other enveloped viruses.
2015-03-10T16:11:55Z
2015-03-10T16:11:55Z
2015-03-01
Article
Mechanisms of methods for hepatitis C virus inactivation. 2015, 81 (5):1616-21 Appl. Environ. Microbiol.
1098-5336
25527548
10.1128/AEM.03580-14
http://hdl.handle.net/10033/346484
Applied and environmental microbiology
en
oai:repository.helmholtz-hzi.de:10033/3465632019-08-30T11:34:22Zcom_10033_620589col_10033_620596
The impact of hepatitis E in the liver transplant setting.
Behrendt, Patrick
Steinmann, Eike
Manns, Michael P
Wedemeyer, Heiner
Hepatitis E virus (HEV) infection has been identified as a cause of graft hepatitis in liver transplant recipients. The true frequency and clinical importance of HEV infections after liver transplantations is a matter of debate. It is proposed that consumption of HEV-contaminated undercooked meat is a main source for HEV infections in developed countries--which might also account for some hepatitis E cases after organ transplantation. However, HEV is also transmitted by transfusion of blood products, likely representing a previously underestimated risk particularly for patients in the transplant setting. HEV infection can take chronic courses in immunocompromised individuals, associated in some cases with rapid progression to cirrhosis within 1-2 years of infection. Diagnosis in transplanted patients is based on HEV RNA testing as antibody assays are not sensitive enough. Selection of immunosuppressive drugs is important as different compounds may influence viral replication and the course of liver disease. Ribavirin has antiviral activity against HEV and should be administered for at least three months in chronically infected individuals; however, treatment failure may occur. HEV infections have also been linked to a variety of extrahepatic manifestations both during and after resolution of infection. In this review we summarize the emerging data on hepatitis E with a particular focus on the importance of HEV infections for liver transplant recipients.
2015-03-12T15:28:20Z
2015-03-12T15:28:20Z
2014-12
Article
The impact of hepatitis E in the liver transplant setting. 2014, 61 (6):1418-29 J. Hepatol.
1600-0641
25195557
10.1016/j.jhep.2014.08.047
http://hdl.handle.net/10033/346563
Journal of hepatology
en
oai:repository.helmholtz-hzi.de:10033/5504862019-08-30T11:25:11Zcom_10033_620589col_10033_620590
Adaptation of hepatitis C virus to mouse CD81 permits infection of mouse cells in the absence of human entry factors.
Bitzegeio, Julia
Bankwitz, Dorothea
Hueging, Kathrin
Haid, Sibylle
Brohm, Christiane
Zeisel, Mirjam B
Herrmann, Eva
Iken, Marcus
Ott, Michael
Baumert, Thomas F
Pietschmann, Thomas
Animals
Antigens, CD
Antigens, CD81
Claudin-1
Hepacivirus
Membrane Proteins
Mice
Receptors, Virus
Scavenger Receptors, Class B
Viral Envelope Proteins
Virus Internalization
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.
2015-04-23T08:12:48Z
2015-04-23T08:12:48Z
2010
Article
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
en
oai:repository.helmholtz-hzi.de:10033/5523172019-08-30T11:25:11Zcom_10033_620589col_10033_620590
Hepatitis C virus p7-a viroporin crucial for virus assembly and an emerging target for antiviral therapy.
Steinmann, Eike
Pietschmann, Thomas
Division of Experimental Virology, Centre for Experimental and Clinical Infection Research, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
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.
2015-05-05T13:52:17Z
2015-05-05T13:52:17Z
2010-09
Article
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
en
oai:repository.helmholtz-hzi.de:10033/5541782019-08-30T11:33:29Zcom_10033_620589col_10033_620590
Control of hepatitis C virus replication in mouse liver-derived cells by MAVS-dependent production of type I and type III interferons.
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.
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.
2015-05-19T15:19:36Z
2015-05-19T15:19:36Z
2015-04
Article
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
en
oai:repository.helmholtz-hzi.de:10033/5795742019-08-30T11:34:48Zcom_10033_620589col_10033_620590
A molecular tweezer antagonizes seminal amyloids and HIV infection.
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.
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.
2015-10-12T13:38:58Z
2015-10-12T13:38:58Z
2015
Article
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
en
oai:repository.helmholtz-hzi.de:10033/5928332019-08-30T11:31:23Zcom_10033_620589col_10033_620596
Host cell mTORC1 is required for HCV RNA replication.
Stöhr, Stefanie
Costa, Rui
Sandmann, Lisa
Westhaus, Sandra
Pfaender, Stephanie
Anggakusuma
Dazert, Eva
Meuleman, Philip
Vondran, Florian W R
Manns, Michael P
Steinmann, Eike
von Hahn, Thomas
Ciesek, Sandra
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.
Chronically HCV-infected orthotopic liver transplantation (OLT) recipients appear to have improved outcomes when their immunosuppressive regimen includes a mammalian target of rapamycin (mTOR) inhibitor. The mechanism underlying this observation is unknown.
2016-01-05T13:52:18Z
2016-01-05T13:52:18Z
2015-08-14
Article
Host cell mTORC1 is required for HCV RNA replication. 2015: Gut
1468-3288
26276683
10.1136/gutjnl-2014-308971
http://hdl.handle.net/10033/592833
Gut
ENG
oai:repository.helmholtz-hzi.de:10033/5932842019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Cell culture-derived HCV cannot infect synovial fibroblasts.
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.
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.
2016-01-11T15:44:25Z
2016-01-11T15:44:25Z
2015
Article
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
en
oai:repository.helmholtz-hzi.de:10033/5963442019-08-30T11:35:14Zcom_10033_620589col_10033_620590
Flunarizine prevents hepatitis C virus membrane fusion in a genotype-dependent manner by targeting the potential fusion peptide within E1.
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.
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.
2016-02-16T14:38:17Z
2016-02-16T14:38:17Z
2016-01
Article
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.)
en
oai:repository.helmholtz-hzi.de:10033/6011702019-08-30T11:33:29Zcom_10033_620589col_10033_620596
Development and virucidal activity of a novel alcohol-based hand disinfectant supplemented with urea and citric acid.
Ionidis, Georgios
Hübscher, Judith
Jack, Thomas
Becker, Britta
Bischoff, Birte
Todt, Daniel
Hodasa, Veronika
Brill, Florian H H
Steinmann, Eike
Steinmann, Jochen
TWINCORE Centre for Experimental and Clinical Infection Research, Feodor-Lynen-Str. 7, 30625, Hannover, Germany.
Hand disinfectants are important for the prevention of virus transmission in the health care system and environment. The development of broad antiviral spectrum hand disinfectants with activity against enveloped and non-enveloped viruses is limited due to a small number of permissible active ingredients able to inactivate viruses.
2016-03-11T14:26:38Z
2016-03-11T14:26:38Z
2016
Article
Development and virucidal activity of a novel alcohol-based hand disinfectant supplemented with urea and citric acid. 2016, 16 (1):77 BMC Infect. Dis.
1471-2334
26864562
10.1186/s12879-016-1410-9
http://hdl.handle.net/10033/601170
BMC infectious diseases
en
oai:repository.helmholtz-hzi.de:10033/6012232019-08-30T11:35:13Zcom_10033_620589col_10033_620590
Distinct Escape Pathway by Hepatitis C Virus Genotype 1a from a Dominant CD8+ T Cell Response by Selection of Altered Epitope Processing.
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.
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.
2016-03-11T15:24:56Z
2016-03-11T15:24:56Z
2015
Article
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
en
oai:repository.helmholtz-hzi.de:10033/6034852019-08-30T11:34:48Zcom_10033_620589col_10033_620596
Natural reservoirs for homologs of hepatitis C virus.
Pfaender, Stephanie
Brown, Richard Jp
Pietschmann, Thomas
Steinmann, Eike
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.
Hepatitis C virus is considered a major public health problem, infecting 2%-3% of the human population. Hepatitis C virus infection causes acute and chronic liver disease, including chronic hepatitis, cirrhosis and hepatocellular carcinoma. In fact, hepatitis C virus infection is the most frequent indication for liver transplantation and a vaccine is not available. Hepatitis C virus displays a narrow host species tropism, naturally infecting only humans, although chimpanzees are also susceptible to experimental infection. To date, there is no evidence for an animal reservoir of viruses closely related to hepatitis C virus which may have crossed the species barrier to cause disease in humans and resulted in the current pandemic. In fact, due to this restricted host range, a robust immunocompetent small animal model is still lacking, hampering mechanistic analysis of virus pathogenesis, immune control and prophylactic vaccine development. Recently, several studies discovered new viruses related to hepatitis C virus, belonging to the hepaci- and pegivirus genera, in small wild mammals (rodents and bats) and domesticated animals which live in close contact with humans (dogs and horses). Genetic and biological characterization of these newly discovered hepatitis C virus-like viruses infecting different mammals will contribute to our understanding of the origins of hepatitis C virus in humans and enhance our ability to study pathogenesis and immune responses using tractable animal models. In this review article, we start with an introduction on the genetic diversity of hepatitis C virus and then focus on the newly discovered viruses closely related to hepatitis C virus. Finally, we discuss possible theories about the origin of this important viral human pathogen.
2016-03-22T12:34:06Z
2016-03-22T12:34:06Z
2014-03
Article
Natural reservoirs for homologs of hepatitis C virus. 2014, 3 (3):e21 Emerg Microbes Infect
2222-1751
26038514
10.1038/emi.2014.19
http://hdl.handle.net/10033/603485
Emerging microbes & infections
en
oai:repository.helmholtz-hzi.de:10033/6062562019-08-30T11:33:05Zcom_10033_620589col_10033_620590
Distinct Escape Pathway by Hepatitis C Virus Genotype 1a from a Dominant CD8+ T Cell Response by Selection of Altered Epitope Processing.
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.
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.
2016-04-21T08:58:39Z
2016-04-21T08:58:39Z
2015
Article
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
en
oai:repository.helmholtz-hzi.de:10033/6051022019-08-30T11:33:05Zcom_10033_620589col_10033_620590
A Diverse Panel of Hepatitis C Virus Glycoproteins for Use in Vaccine Research Reveals Extremes of Monoclonal Antibody Neutralization Resistance.
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.
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.
2016-04-12T14:35:11Z
2016-04-12T14:35:11Z
2015
Article
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
en
oai:repository.helmholtz-hzi.de:10033/6072262019-08-30T11:34:48Zcom_10033_620589col_10033_620596
Frequent presence of hepaci and pegiviruses in commercial equine serum pools.
Postel, Alexander
Cavalleri, Jessika-M V
Pfaender, Stephanie
Walter, Stephanie
Steinmann, E
Fischer, Nicole
Feige, Karsten
Haas, Ludwig
Becher, Paul
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.
Novel viruses belonging to the genera Hepacivirus and Pegivirus have recently been discovered in horses and other animal species. Viral genomes of non-primate hepaciviruses (NPHV), equine pegivirus 1 (EPgV 1) and Theiler's disease associated virus (TDAV) were detected in a horse serum routinely used for cell culture propagation in our laboratory. Therefore, a study was carried out to further investigate the presence of these human Hepatitis C virus (HCV) related viruses in equine serum based products used in veterinary medicine and for research and to characterize the viral genomes. Without exception all commercially available equine sera purchased for cell culture propagation (n=6) were tested positive for NPHV, EPgV 1 or TDAV genomes by qRT-PCR. Molecular analyses of one single commercial horse serum from Europe confirmed multiple viral genomes, including two TDAV genomes significantly different from the only published TDAV sequence. Furthermore, multiple batches of horse serum pools (n=35) collected for manufacturing of biological products turned out to be positive for NPHV and EPgV 1 genomes. Nevertheless, the final commercial products (n=9) were exclusively tested qRT-PCR negative. Field samples (n=119) obtained from two premises located in the same region as the donor horses were analyzed, demonstrating the frequent presence of NPHV and EPgV 1, but the absence of TDAV genomes. The presence of NPHV, EPgV 1 and TDAV in commercial equine sera and serum based products could have considerable consequences for biosecurity and may also bias the outcome of research studies conducted with related viruses.
2016-04-27T14:27:42Z
2016-04-27T14:27:42Z
2016-01-15
Article
Frequent presence of hepaci and pegiviruses in commercial equine serum pools. 2016, 182:8-14 Vet. Microbiol.
1873-2542
26711022
10.1016/j.vetmic.2015.10.032
http://hdl.handle.net/10033/607226
Veterinary microbiology
en
oai:repository.helmholtz-hzi.de:10033/6077362019-08-30T11:33:05Zcom_10033_620589col_10033_620590
ABHD5/CGI-58, the Chanarin-Dorfman Syndrome Protein, Mobilises Lipid Stores for Hepatitis C Virus Production.
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.
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.
2016-05-03T13:15:16Z
2016-05-03T13:15:16Z
2016-04
Article
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
en
oai:repository.helmholtz-hzi.de:10033/6090552019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Hepatic differentiation of human pluripotent stem cells in miniaturized format suitable for high-throughput screen.
Carpentier, Arnaud
Nimgaonkar, Ila
Chu, Virginia
Xia, Yuchen
Hu, Zongyi
Liang, T Jake
Twincore, Institute for Experimental Virology, Hannover, Germany.
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.
2016-05-11T14:20:35Z
2016-05-11T14:20:35Z
2016-03-29
Article
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
ENG
oai:repository.helmholtz-hzi.de:10033/6112382019-08-30T11:24:31Zcom_10033_620589col_10033_620595
Cellular Antiviral Factors that Target Particle Infectivity of HIV-1.
Goffinet, Christine
TWINCORE, Zentrum für Experimentelle und Klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
In the past decade, the identification and characterization of antiviral genes with the ability to interfere with virus replication has established cell-intrinsic innate immunity as a third line of antiviral defense in addition to adaptive and classical innate immunity. Understanding how cellular factors have evolved to inhibit HIV-1 reveals particularly vulnerable points of the viral replication cycle. Many, but not all, antiviral proteins share type I interferon-upregulated expression and sensitivity to viral counteraction or evasion measures. Whereas well-established restriction factors interfere with early post-entry steps and release of HIV-1, recent research has revealed a diverse set of proteins that reduce the infectious quality of released particles using individual, to date poorly understood modes of action. These include induction of paucity of mature glycoproteins in nascent virions or self-incorporation into the virus particle, resulting in poor infectiousness of the virion and impaired spread of the infection.
2016-05-31T13:07:52Z
2016-05-31T13:07:52Z
2016
Article
Cellular Antiviral Factors that Target Particle Infectivity of HIV-1. 2016, 14 (3):211-6 Curr. HIV Res.
1873-4251
26674651
http://hdl.handle.net/10033/611238
Current HIV research
en
oai:repository.helmholtz-hzi.de:10033/6116972019-08-30T11:33:30Zcom_10033_620589col_10033_620590
Several Human Liver Cell Expressed Apolipoproteins Complement HCV Virus Production with Varying Efficacy Conferring Differential Specific Infectivity to Released Viruses.
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.
Apolipoproteins
Cell Line
Hepacivirus
Hepatocytes
Humans
Virulence
Virus Replication
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.
2016-06-03T09:22:58Z
2016-06-03T09:22:58Z
2015
Article
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
en
oai:repository.helmholtz-hzi.de:10033/6140652019-08-30T11:27:46Zcom_10033_620589col_10033_620596
Echinocandin resistance and population structure of invasive Candida glabrata isolates from two university hospitals in Germany and Austria.
Klotz, Ulrike
Schmidt, Dirk
Willinger, Birgit
Steinmann, Eike
Buer, Jan
Rath, Peter-Michael
Steinmann, Joerg
Twincore Centre for Experimental and Clinical Infection Research, Hannover, Germany.
Echinocandin resistance in Candida glabrata is emerging and is associated with the presence of FKS mutations. In this study, we analysed the antifungal susceptibility, presence of FKS mutations and clonality of C. glabrata blood culture isolates from two hospitals in Germany and Austria. Susceptibility testing of 64 C. glabrata bloodstream isolates from two university hospitals was performed with broth microdilution method according to EUCAST. In addition, all isolates were screened for FKS mutations. Molecular fingerprinting was performed by microsatellite PCR with three separate primer pairs and semiautomated repetitive sequenced-based PCR (rep-PCR). One C. glabrata isolate from Germany (1.5%) was echinocandin resistant, with a corresponding mutation in FKS2 gene hot spot 1. The discriminatory power of microsatellite PCR was higher than that of rep-PCR (Simpson Index of 0.94 vs. 0.88); microsatellite PCR created 31 separate genotypes, whereas rep-PCR created 17. Predominant genotypes or clusters of isolates from Germany and Austria were present, with no epidemiological evidence of nosocomial transmissions. Although we found a low incidence of echinocandin resistance in C. glabrata in our settings, further surveillance projects in central Europe are warranted for monitoring future epidemiological trends. The genetic population structure of C. glabrata demonstrates overrepresented geographical clusters.
2016-06-22T12:04:00Z
2016-06-22T12:04:00Z
2016-05
Article
Echinocandin resistance and population structure of invasive Candida glabrata isolates from two university hospitals in Germany and Austria. 2016, 59 (5):312-8 Mycoses
1439-0507
26806376
10.1111/myc.12472
http://hdl.handle.net/10033/614065
Mycoses
en
oai:repository.helmholtz-hzi.de:10033/6158132019-08-30T11:34:22Zcom_10033_620589col_10033_620590
Decoding protein networks during virus entry by quantitative proteomics.
Gerold, Gisa
Bruening, Janina
Pietschmann, Thomas
TWINCORE, Centre for Experimental and Clinical Infection Research, Feodor-Lynen-Str. 7 30625 Hannover, Germany.
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.
2016-07-08T14:31:39Z
2016-07-08T14:31:39Z
2016-06-15
Article
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
en
info:eu-repo/grantAgreement/EC/FP7/281473
openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6169242019-08-30T11:35:13Zcom_10033_620589col_10033_620590
Hepatitis C Virus Stimulates Murine CD8α-Like Dendritic Cells to Produce Type I Interferon in a TRIF-Dependent Manner.
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.
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.
2016-07-14T08:05:16Z
2016-07-14T08:05:16Z
2016-07
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6190352019-08-30T11:35:13Zcom_10033_620589col_10033_620596
Extra-hepatic replication and infection of hepatitis E virus in neuronal-derived cells.
Drave, S A
Debing, Y
Walter, S
Todt, D
Engelmann, M
Friesland, M
Wedemeyer, H
Neyts, J
Behrendt, P
Steinmann, E
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.
Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and a member of the genus Orthohepevirus in the family Hepeviridae. Infection usually leads to acute hepatitis that can become fulminant, particularly among pregnant women and in patients with preexisting liver disease, or may evolve to a chronic state, especially in immunosuppressed individuals. HEV has been shown to produce a range of extra-hepatic manifestations including aplastic anaemia, acute thyroiditis, glomerulonephritis as well as neurological disorders such as Guillain-Barré syndrome, neuralgic amyotrophy and encephalitis. The pathogenesis of these neurological injuries remains largely unknown, and it is also uncertain whether or not HEV can directly infect neuronal cells. In this study, we investigated whether HEV is capable of completing the viral life cycle in human neuronal-derived cell lines such as neuroepithelioma (SK-N-MC), desmoplastic cerebellar medulloblastoma (DAOY), glioblastoma multiforme (DBTRG), glioblastoma astrocytoma (U-373 MG) and oligodendrocytic (M03.13) cells. Following transfection of these cells with HEV Gaussia luciferase reporter virus, all tested cell lines supported HEV RNA replication. Furthermore, extra- and intracellular viral capsid was detected by an HEV antigen ELISA as a marker for virus assembly and release. Permissiveness for HEV cell entry could be demonstrated for the oligodendrocytic cell line M03.13. In conclusion, these results indicate that HEV tropism is not restricted to the liver and HEV can potentially complete the full viral life cycle in neuronal-derived tissues explaining neurologic disorders during HEV infection.
2016-08-30T08:51:02Z
2016-08-30T08:51:02Z
2016-07
Article
Extra-hepatic replication and infection of hepatitis E virus in neuronal-derived cells. 2016, 23 (7):512-21 J. Viral Hepat.
1365-2893
26891712
10.1111/jvh.12515
http://hdl.handle.net/10033/619035
Journal of viral hepatitis
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6205442019-08-30T11:36:04Zcom_10033_620589col_10033_620590
Hepacivirus NS3/4A proteases interfere with MAVS signalling of their cognate animal hosts and also with human MAVS: implications for zoonotic transmission.
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.
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.
2016-10-07T14:33:24Z
2016-10-07T14:33:24Z
2016-09-21
Article
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
ENG
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6205492019-08-30T11:35:39Zcom_10033_620589col_10033_620590
Ebola Virus Disease Is Characterized by Poor Activation and Reduced Levels of Circulating CD16+ Monocytes.
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.
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.
2016-10-12T13:56:47Z
2016-10-12T13:56:47Z
2016-10-15
Article
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
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/
oai:repository.helmholtz-hzi.de:10033/6205532019-08-30T11:33:53Zcom_10033_620589col_10033_620590
Hepatitis C virus plays hide and seek with neutralizing antibodies.
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
Article
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/
oai:repository.helmholtz-hzi.de:10033/6205662019-08-30T11:30:58Zcom_10033_620589col_10033_620596
Emergence of linezolid- and vancomycin-resistant Enterococcus faecium in a department for hematologic stem cell transplantation.
Krull, M
Klare, I
Ross, B
Trenschel, R
Beelen, D W
Todt, D
Steinmann, E
Buer, J
Rath, P-M
Steinmann, J
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.
Prevalence of vancomycin-resistant enterococci has increased in Germany. Here, we report the cluster of linezolid- and vancomycin-resistant Enterococcus faecium (LVRE) in a German department for hematologic stem cell transplantation (HSCT).
2016-11-02T09:30:57Z
2016-11-02T09:30:57Z
2016
Article
Emergence of linezolid- and vancomycin-resistant Enterococcus faecium in a department for hematologic stem cell transplantation. 2016, 5:31 Antimicrob Resist Infect Control
27688876
10.1186/s13756-016-0131-6
http://hdl.handle.net/10033/620566
Antimicrobial resistance and infection control
ENG
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6205832019-08-30T11:26:13Zcom_10033_620589col_10033_620590
Unique human immune signature of Ebola virus disease in Guinea.
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.
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
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.
2016-11-17T12:19:23Z
2016-11-17T12:19:23Z
2016-05-05
Article
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
ENG
: info:eu-repo/grantAgreement/H2020/666100
openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6205842019-08-30T11:35:13Zcom_10033_620589col_10033_620596
Mutagenic Effects of Ribavirin on Hepatitis E Virus-Viral Extinction versus Selection of Fitness-Enhancing Mutations.
Todt, Daniel
Walter, Stephanie
Brown, Richard J P
Steinmann, Eike
TWINCORE, Centre for experimental and clinical infection research GmbH, Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Hepatitis E virus (HEV), an important agent of viral hepatitis worldwide, can cause severe courses of infection in pregnant women and immunosuppressed patients. To date, HEV infections can only be treated with ribavirin (RBV). Major drawbacks of this therapy are that RBV is not approved for administration to pregnant women and that the virus can acquire mutations, which render the intra-host population less sensitive or even resistant to RBV. One of the proposed modes of action of RBV is a direct mutagenic effect on viral genomes, inducing mismatches and subsequent nucleotide substitutions. These transition events can drive the already error-prone viral replication beyond an error threshold, causing viral population extinction. In contrast, the expanded heterogeneous viral population can facilitate selection of mutant viruses with enhanced replication fitness. Emergence of these mutant viruses can lead to therapeutic failure. Consequently, the onset of RBV treatment in chronically HEV-infected individuals can result in two divergent outcomes: viral extinction versus selection of fitness-enhanced viruses. Following an overview of RNA viruses treated with RBV in clinics and a summary of the different antiviral modes of action of this drug, we focus on the mutagenic effect of RBV on HEV intrahost populations, and how HEV is able to overcome lethal mutagenesis.
2016-11-17T15:30:34Z
2016-11-17T15:30:34Z
2016-10-13
Article
Mutagenic Effects of Ribavirin on Hepatitis E Virus-Viral Extinction versus Selection of Fitness-Enhancing Mutations. 2016, 8 (10) Viruses
1999-4915
27754363
10.3390/v8100283
http://hdl.handle.net/10033/620584
Viruses
ENG
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6206242019-08-30T11:26:42Zcom_10033_620589col_10033_620596
Inactivation of HCV and HIV by microwave: a novel approach for prevention of virus transmission among people who inject drugs.
Siddharta, Anindya
Pfaender, Stephanie
Malassa, Angelina
Doerrbecker, Juliane
Anggakusuma
Engelmann, Michael
Nugraha, Boya
Steinmann, Joerg
Todt, Daniel
Vondran, Florian W R
Mateu-Gelabert, Pedro
Goffinet, Christine
Steinmann, Eike
Hepatitis C virus (HCV) and human immunodeficiency virus (HIV-1) transmissions among people who inject drugs (PWID) continue to pose a challenging global health problem. Here, we aimed to analyse a universally applicable inactivation procedure, namely microwave irradiation, as a safe and effective method to reduce the risk of viral transmission. The exposure of HCV from different genotypes to microwave irradiation resulted in a significant reduction of viral infectivity. Furthermore, microwave irradiation reduced viral infectivity of HIV-1 and of HCV/HIV-1 suspensions indicating that this inactivation may be effective at preventing co-infections. To translate microwave irradiation as prevention method to used drug preparation equipment, we could further show that HCV as well as HIV-1 infectivity could be abrogated in syringes and filters. This study demonstrates the power of microwave irradiation for the reduction of viral transmission and establishment of this safety strategy could help reduce the transmission of blood-borne viruses.
2016-12-05T15:14:17Z
2016-12-05T15:14:17Z
2016-11-18
Article
Inactivation of HCV and HIV by microwave: a novel approach for prevention of virus transmission among people who inject drugs. 2016, 6:36619 Sci Rep
2045-2322
27857152
10.1038/srep36619
http://hdl.handle.net/10033/620624
Scientific reports
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6206302019-08-30T11:34:48Zcom_10033_620589col_10033_620596
Inactivation of Zika virus in human breast milk by prolonged storage or pasteurization.
Pfaender, Stephanie
Vielle, Nathalie J
Ebert, Nadine
Steinmann, Eike
Alves, Marco P
Thiel, Volker
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.
Zika virus infection during pregnancy poses a serious risk for pregnant women as it can cause severe birth defects. Even though the virus is mainly transmitted via mosquitos, human-to-human transmission has been described. Infectious viral particles have been detected in breast milk of infected women which raised concerns regarding the safety of breastfeeding in areas of Zika virus transmission or in case of a suspected or confirmed Zika virus infection. In this study, we show that Zika virus is effectively inactivated in human breast milk after prolonged storage or upon pasteurization of milk.
2016-12-06T10:05:13Z
2016-12-06T10:05:13Z
2016-11-23
Article
Inactivation of Zika virus in human breast milk by prolonged storage or pasteurization. 2016, 228:58-60 Virus Res.
1872-7492
27889615
10.1016/j.virusres.2016.11.025
http://hdl.handle.net/10033/620630
Virus research
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6206642019-08-30T11:37:23Zcom_10033_620589col_10033_620596
Acute and chronic infections with nonprimate hepacivirus in young horses.
Gather, Theresa
Walter, Stephanie
Pfaender, Stephanie
Todt, Daniel
Feige, Karsten
Steinmann, Eike
Cavalleri, Jessika M V
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.
The recently discovered nonprimate hepacivirus (NPHV) naturally infects horses and is the closest known homolog of hepatitis C virus to date. Within a follow-up study acute field infections were monitored in four young Thoroughbred horses until the ages of 12-13 months. Serum samples were analyzed for the presence of NPHV RNA and anti-NPHV NS3 antibodies and liver specific parameters were evaluated. The four young horses were not able to clear infection, but remained chronically infected for the entire monitored time period despite the presence of NPHV specific antibodies.
2016-12-14T09:20:28Z
2016-12-14T09:20:28Z
2016-09-22
Article
Acute and chronic infections with nonprimate hepacivirus in young horses. 2016, 47 (1):97 Vet. Res.
1297-9716
27659317
10.1186/s13567-016-0381-6
http://hdl.handle.net/10033/620664
Veterinary research
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6206902019-08-30T11:35:10Zcom_10033_620589col_10033_620596
Apolipoprotein E polymorphisms and their protective effect on hepatitis E virus replication.
Weller, Romy
Todt, Daniel
Engelmann, Michael
Friesland, Martina
Wedemeyer, Heiner
Pietschmann, Thomas
Steinmann, Eike
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.
2017-01-10T12:53:21Z
2017-01-10T12:53:21Z
2016-12
Other
Apolipoprotein E polymorphisms and their protective effect on hepatitis E virus replication. 2016, 64 (6):2274-2276 Hepatology
1527-3350
27541341
10.1002/hep.28788
http://hdl.handle.net/10033/620690
Hepatology (Baltimore, Md.)
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6208042019-08-30T11:36:32Zcom_10033_620589col_10033_620596
In vivo evidence for ribavirin-induced mutagenesis of the hepatitis E virus genome.
Todt, Daniel
Gisa, Anett
Radonic, Aleksandar
Nitsche, Andreas
Behrendt, Patrick
Suneetha, Pothakamuri Venkata
Pischke, Sven
Bremer, Birgit
Brown, Richard J P
Manns, Michael P
Cornberg, Markus
Bock, C Thomas
Steinmann, Eike
Wedemeyer, Heiner
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.
Hepatitis E virus (HEV) infection can take chronic courses in immunocompromised patients potentially leading to liver cirrhosis and liver failure. Ribavirin (RBV) is currently the only treatment option for many patients, but treatment failure can occur which has been associated with the appearance of a distinct HEV polymerase mutant (G1634R). Here, we performed a detailed analysis of HEV viral intrahost evolution during chronic hepatitis E infections.
2017-02-03T09:50:39Z
2017-02-03T09:50:39Z
2016-10
Article
In vivo evidence for ribavirin-induced mutagenesis of the hepatitis E virus genome. 2016, 65 (10):1733-43 Gut
1468-3288
27222534
10.1136/gutjnl-2015-311000
http://hdl.handle.net/10033/620804
Gut
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6208502019-08-30T11:34:47Zcom_10033_620589col_10033_620596
Successful retreatment of a patient with chronic hepatitis C genotype 2k/1b virus with ombitasvir/paritaprevir/ritonavir plus dasabuvir.
Todt, Daniel
Schlevogt, Bernhard
Deterding, Katja
Grundhoff, Adam
Manns, Michael P
Wedemeyer, Heiner
Fischer, Nicole
Cornberg, Markus
Steinmann, Eike
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.
2017-03-07T09:40:41Z
2017-03-07T09:40:41Z
2017-01-18
Article
Successful retreatment of a patient with chronic hepatitis C genotype 2k/1b virus with ombitasvir/paritaprevir/ritonavir plus dasabuvir. 2017 J. Antimicrob. Chemother.
1460-2091
28100444
10.1093/jac/dkw572
http://hdl.handle.net/10033/620850
The Journal of antimicrobial chemotherapy
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6208522019-08-30T11:35:14Zcom_10033_620589col_10033_620596
Biofilm formation of the black yeast-like fungus Exophiala dermatitidis and its susceptibility to antiinfective agents.
Kirchhoff, Lisa
Olsowski, Maike
Zilmans, Katrin
Dittmer, Silke
Haase, Gerhard
Sedlacek, Ludwig
Steinmann, Eike
Buer, Jan
Rath, Peter-Michael
Steinmann, Joerg
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.
Various fungi have the ability to colonize surfaces and to form biofilms. Fungal biofilm-associated infections are frequently refractory to targeted treatment because of resistance to antifungal drugs. One fungus that frequently colonises the respiratory tract of cystic fibrosis (CF) patients is the opportunistic black yeast-like fungus Exophiala dermatitidis. We investigated the biofilm-forming ability of E. dermatitidis and its susceptibility to various antiinfective agents and natural compounds. We tested 58 E. dermatitidis isolates with a biofilm assay based on crystal violet staining. In addition, we used three isolates to examine the antibiofilm activity of voriconazole, micafungin, colistin, farnesol, and the plant derivatives 1,2,3,4,6-penta-O-galloyl-b-D-glucopyranose (PGG) and epigallocatechin-3-gallate (EGCG) with an XTT reduction assay. We analysed the effect of the agents on cell to surface adhesion, biofilm formation, and the mature biofilm. The biofilms were also investigated by confocal laser scan microscopy. We found that E. dermatitidis builds biofilm in a strain-specific manner. Invasive E. dermatitidis isolates form most biomass in biofilm. The antiinfective agents and the natural compounds exhibited poor antibiofilm activity. The greatest impact of the compounds was detected when they were added prior cell adhesion. These findings suggest that prevention may be more effective than treatment of biofilm-associated E. dermatitidis infections.
2017-03-08T10:30:17Z
2017-03-08T10:30:17Z
2017-02-17
Article
Biofilm formation of the black yeast-like fungus Exophiala dermatitidis and its susceptibility to antiinfective agents. 2017, 7:42886 Sci Rep
2045-2322
28211475
10.1038/srep42886
http://hdl.handle.net/10033/620852
Scientific reports
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6208592019-08-30T11:29:17Zcom_10033_620589col_10033_620596
Prevention strategies for blood-borne viruses-in the Era of vaccines, direct acting antivirals and antiretroviral therapy.
Pfaender, Stephanie
von Hahn, Thomas
Steinmann, Joerg
Ciesek, Sandra
Steinmann, Eike
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.
Blood-borne viruses, such as hepatitis B virus, hepatitis C virus, human immunodeficiency virus, and the facultative blood-borne hepatitis E virus, are considered a major public health problem given that they are accountable for millions of deaths each year. Treatment options, including effective vaccine design, development of antiviral strategies and the implementation of antiretroviral therapy have improved substantially over the last couple of years and contribute to successful treatment and prevention of these infectious diseases. In this review, we summarise the current knowledge and concepts in prevention of transmission of these blood-borne viruses.
2017-03-15T09:04:49Z
2017-03-15T09:04:49Z
2016-09
Article
Prevention strategies for blood-borne viruses-in the Era of vaccines, direct acting antivirals and antiretroviral therapy. 2016, 26 (5):330-9 Rev. Med. Virol.
1099-1654
27185010
10.1002/rmv.1890
http://hdl.handle.net/10033/620859
Reviews in medical virology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6209012019-08-30T11:33:29Zcom_10033_620589col_10033_620590
Clinically Approved Ion Channel Inhibitors Close Gates for Hepatitis C Virus and Open Doors for Drug Repurposing in Infectious Viral Diseases.
Pietschmann, Thomas
TwinCore, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen Str. 7, 30625 Hannover, Germany.
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.
2017-04-21T10:35:55Z
2017-04-21T10:35:55Z
2017-01-15
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6209362019-08-30T11:33:57Zcom_10033_620589col_10033_620590
Different features of Vδ2 T and NK cells in fatal and non-fatal human Ebola infections.
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.
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.
2017-06-09T08:50:46Z
2017-06-09T08:50:46Z
2017-05-30
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6209732019-08-30T11:34:48Zcom_10033_620589col_10033_620590col_10033_620596
Virucidal Activity of World Health Organization-Recommended Formulations Against Enveloped Viruses, Including Zika, Ebola, and Emerging Coronaviruses.
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.
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.
2017-06-22T09:40:09Z
2017-06-22T09:40:09Z
2017-03-15
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6209812019-08-30T11:34:48Zcom_10033_620589col_10033_620590
Maturation of secreted HCV particles by incorporation of secreted ApoE protects from antibodies by enhancing infectivity.
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.
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.
2017-06-26T14:37:58Z
2017-06-26T14:37:58Z
2017-04-22
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6210092019-08-30T11:28:51Zcom_10033_620589com_10033_620636col_10033_620590col_10033_620596col_10033_620638
Tracking HCV protease population diversity during transmission and susceptibility of founder populations to antiviral therapy.
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.
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.
2017-07-12T14:20:02Z
2017-07-12T14:20:02Z
2017-03
Article
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
en
info:eu-repo/grantAgreement/EC/FP7/305600
openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6210102019-08-30T11:36:32Zcom_10033_620601com_10033_620589col_10033_620602col_10033_620590col_10033_620590
Immune protection against reinfection with nonprimate hepacivirus.
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.
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.
2017-07-13T11:52:32Z
2017-07-13T11:52:32Z
2017-03-21
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6210772019-08-30T11:36:33Zcom_10033_620589com_10033_311308col_10033_620721col_10033_620590col_10033_620596
Assessment of cross-species transmission of hepatitis C virus-related non-primate hepacivirus in a population of humans at high risk of exposure.
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.
Adult
Agricultural Workers' Diseases
Animals
Female
Hepacivirus
Hepatitis C
Horse Diseases
Horses
Humans
Male
Middle Aged
Occupational Exposure
Phylogeny
Zoonoses
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.
2017-08-29T08:45:06Z
2017-08-29T08:45:06Z
2015-09
Article
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
en
info:eu-repo/grantAgreement/EC/FP7/281473
openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6211322019-08-30T11:33:51Zcom_10033_620589col_10033_620595
[cGAS, an antiviral weapon: role in the recognition of HIV-1 transmitted from cell to cell].
Ducroux, Aurélie
Goffinet, Christine
TwinCore, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
2017-10-10T13:09:31Z
2017-10-10T13:09:31Z
2017-10-10
Article
[cGAS, an antiviral weapon: role in the recognition of HIV-1 transmitted from cell to cell]., 33 (8-9):732-734 Med Sci (Paris)
1958-5381
28945560
10.1051/medsci/20173308015
http://hdl.handle.net/10033/621132
Medecine sciences : M/S
french
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6211592019-08-30T11:27:46Zcom_10033_620589col_10033_620596
Differential Infection Patterns and Recent Evolutionary Origins of Equine Hepaciviruses in Donkeys.
Walter, Stephanie
Rasche, Andrea
Moreira-Soto, Andrés
Pfaender, Stephanie
Bletsa, Magda
Corman, Victor Max
Aguilar-Setien, Alvaro
García-Lacy, Fernando
Hans, Aymeric
Todt, Daniel
Schuler, Gerhard
Shnaiderman-Torban, Anat
Steinman, Amir
Roncoroni, Cristina
Veneziano, Vincenzo
Rusenova, Nikolina
Sandev, Nikolay
Rusenov, Anton
Zapryanova, Dimitrinka
García-Bocanegra, Ignacio
Jores, Joerg
Carluccio, Augusto
Veronesi, Maria Cristina
Cavalleri, Jessika M V
Drosten, Christian
Lemey, Philippe
Steinmann, Eike
Drexler, Jan Felix
TwinCore, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str.7, 30625 Hannover, Germany.
Acute Disease
Animals
Antibodies, Viral
Biological Evolution
Equidae
Europe
Genetic Variation
Genome, Viral
Hepacivirus
Hepatitis C
Horses
Host Specificity
Humans
Israel
Kenya
Latin America
Phylogeny
Sequence Analysis, DNA
Seroepidemiologic Studies
The hepatitis C virus (HCV) is a major human pathogen. Genetically related viruses in animals suggest a zoonotic origin of HCV. The closest relative of HCV is found in horses (termed equine hepacivirus [EqHV]). However, low EqHV genetic diversity implies relatively recent acquisition of EqHV by horses, making a derivation of HCV from EqHV unlikely. To unravel the EqHV evolutionary history within equid sister species, we analyzed 829 donkeys and 53 mules sampled in nine European, Asian, African, and American countries by molecular and serologic tools for EqHV infection. Antibodies were found in 278 animals (31.5%), and viral RNA was found in 3 animals (0.3%), all of which were simultaneously seropositive. A low RNA prevalence in spite of high seroprevalence suggests a predominance of acute infection, a possible difference from the mostly chronic hepacivirus infection pattern seen in horses and humans. Limitation of transmission due to short courses of infection may explain the existence of entirely seronegative groups of animals. Donkey and horse EqHV strains were paraphyletic and 97.5 to 98.2% identical in their translated polyprotein sequences, making virus/host cospeciation unlikely. Evolutionary reconstructions supported host switches of EqHV between horses and donkeys without the involvement of adaptive evolution. Global admixture of donkey and horse hepaciviruses was compatible with anthropogenic alterations of EqHV ecology. In summary, our findings do not support EqHV as the origin of the significantly more diversified HCV. Identification of a host system with predominantly acute hepacivirus infection may enable new insights into the chronic infection pattern associated with HCV.
2017-11-03T15:05:53Z
2017-11-03T15:05:53Z
2017-01-01
Article
Differential Infection Patterns and Recent Evolutionary Origins of Equine Hepaciviruses in Donkeys. 2017, 91 (1) J. Virol.
1098-5514
27795428
10.1128/JVI.01711-16
http://hdl.handle.net/10033/621159
Journal of virology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6211862019-08-30T11:32:41Zcom_10033_620589col_10033_620596
Virucidal efficacy of peracetic acid for instrument disinfection.
Becker, Britta
Brill, Florian H H
Todt, Daniel
Steinmann, Eike
Lenz, Johannes
Paulmann, Dajana
Bischoff, Birte
Steinmann, Jochen
TwinCore, Zentrum für experimentelle und klinische Infektionsforschng GmbH, Feodor-Lynen-Str.7, 30625 Hannover, Germany.
Various peracetic-acid (PAA)-based products for processing flexible endoscopes on the market are often based on a two-component system including a cleaning step before the addition of PAA as disinfectant. The peracetic acid concentrations in these formulations from different manufacturers are ranging from 400 to 1500 ppm (part per million). These products are used at temperatures between 20 °C and 37 °C. Since information on the virus-inactivating properties of peracetic acid at different concentrations and temperature is missing, it was the aim of the study to evaluate peracetic acid solutions against test viruses using the quantitative suspension test, EN 14476. In addition, further studies were performed with the recently established European pre norm (prEN 17111:2017) describing a carrier assay for simulating practical conditions using frosted glass.
2017-11-29T09:28:17Z
2017-11-29T09:28:17Z
2017
Article
Virucidal efficacy of peracetic acid for instrument disinfection. 2017, 6:114 Antimicrob Resist Infect Control
2047-2994
29177047
10.1186/s13756-017-0271-3
http://hdl.handle.net/10033/621186
Antimicrobial resistance and infection control
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6212062019-08-30T11:26:13Zcom_10033_620589col_10033_620595
Potent and reversible lentiviral vector restriction in murine induced pluripotent stem cells.
Geis, Franziska K
Galla, Melanie
Hoffmann, Dirk
Kuehle, Johannes
Zychlinski, Daniela
Maetzig, Tobias
Schott, Juliane W
Schwarzer, Adrian
Goffinet, Christine
Goff, Stephen P
Schambach, Axel
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
Retroviral vectors are derived from wild-type retroviruses, can be used to study retrovirus-host interactions and are effective tools in gene and cell therapy. However, numerous cell types are resistant or less permissive to retrovirus infection due to the presence of active defense mechanisms, or the absence of important cellular host co-factors. In contrast to multipotent stem cells, pluripotent stem cells (PSC) have potential to differentiate into all three germ layers. Much remains to be elucidated in the field of anti-viral immunity in stem cells, especially in PSC.
2017-12-13T15:14:28Z
2017-12-13T15:14:28Z
2017-05-31
Article
Potent and reversible lentiviral vector restriction in murine induced pluripotent stem cells. 2017, 14 (1):34 Retrovirology
1742-4690
28569216
10.1186/s12977-017-0358-1
http://hdl.handle.net/10033/621206
Retrovirology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6212292019-08-30T11:33:57Zcom_10033_620589col_10033_620590
The Role of Type III Interferons in Hepatitis C Virus Infection and Therapy.
Bruening, Janina
Weigel, Bettina
Gerold, Gisa
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
Animals
Antiviral Agents
Hepacivirus
Hepatitis C
Host-Pathogen Interactions
Humans
Immunity, Innate
Interferons
Interleukins
Liver
Signal Transduction
Virus Replication
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.
2018-01-09T15:25:20Z
2018-01-09T15:25:20Z
2017
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6212572019-08-30T11:28:23Zcom_10033_620589col_10033_620590
Protein Interactions during the Flavivirus and Hepacivirus Life Cycle.
Gerold, Gisa
Bruening, Janina
Weigel, Bettina
Pietschmann, Thomas
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
Flavivirus
Flavivirus Infections
Genome, Viral
Hepacivirus
Hepatitis C
Host-Pathogen Interactions
Humans
Protein Interaction Maps
Virus Internalization
Virus Replication
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.
2018-01-29T13:53:38Z
2018-01-29T13:53:38Z
2017-04
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6212612019-08-30T11:35:13Zcom_10033_620589col_10033_620590
Axl can serve as entry factor for Lassa virus depending on the functional glycosylation of dystroglycan.
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.
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.
2018-01-30T15:17:34Z
2018-01-30T15:17:34Z
2017-12-13
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6213052019-08-30T11:26:13Zcom_10033_620636com_10033_620589col_10033_620665col_10033_620590
Type I Interferons Interfere with the Capacity of mRNA Lipoplex Vaccines to Elicit Cytolytic T Cell Responses.
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.
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
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.
2018-03-06T09:44:42Z
2018-03-06T09:44:42Z
2016-11
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6213282019-08-30T11:34:22Zcom_10033_620857com_10033_620589com_10033_620618col_10033_620858col_10033_620622col_10033_620596
Two New Cyathane Diterpenoids from Mycelial Cultures of the Medicinal Mushroom Hericium erinaceus and the Rare Species, Hericium flagellum.
Rupcic, Zeljka
Rascher, Monique
Kanaki, Sae
Köster, Reinhard W
Stadler, Marc
Wittstein, Kathrin
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Basidiomycetes of the genusHericiumare among the most praised medicinal and edible mushrooms, which are known to produce secondary metabolites with the potential to treat neurodegenerative diseases. This activity has been attributed to the discovery of various terpenoids that can stimulate the production of nerve growth factor (NGF) or (as established more recently) brain-derived neurotrophic factor (BDNF) in cell-based bioassays. The present study reports on the metabolite profiles of a Lion's Mane mushroom (Hericium erinaceus) strain and a strain of the rare species,Hericium flagellum(synonymH. alpestre). While we observed highly similar metabolite profiles between the two strains that were examined, we isolated two previously undescribed metabolites, given the trivial names erinacines Z1 and Z2. Their chemical structures were elucidated by means of nuclear magnetic resonance (NMR) spectroscopy and high resolution mass spectrometry. Along with six further, previously identified cyathane diterpenes, the novel erinacines were tested for neurotrophin inducing effects. We found that erinacines act onBDNF, which is a neurotrophic factor that has been reported recently by us to be induced by the corallocins, but as well onNGFexpression, which is consistent with the literature.
2018-03-21T11:47:18Z
2018-03-21T11:47:18Z
2018-03-06
Article
Two New Cyathane Diterpenoids from Mycelial Cultures of the Medicinal Mushroom Hericium erinaceus and the Rare Species, Hericium flagellum. 2018, 19 (3) Int J Mol Sci
1422-0067
29509661
10.3390/ijms19030740
http://hdl.handle.net/10033/621328
International journal of molecular sciences
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6213292019-08-30T11:31:23Zcom_10033_620857com_10033_620589com_10033_620618col_10033_620858col_10033_620622col_10033_620596
Six Heterocyclic Metabolites from the Myxobacterium Labilithrix luteola.
Mulwa, Lucky S
Jansen, Rolf
Praditya, Dimas F
Mohr, Kathrin I
Wink, Joachim
Steinmann, Eike
Stadler, Marc
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Two new secondary metabolites, labindole A [2-methyl-3-(2-nitroethyl)-3H-indole] (1) and labindole B [2-methyl-3-(2-nitrovinyl)-3H-indole] (2), were isolated from the myxobacteriumLabilithrixluteola(DSM 27648T). Additionally, four metabolites3,4,5and6already known from other sources were obtained. Their structures were elucidated from high resolution electrospray ionisation mass spectrometry (HRESIMS) and 1D and 2D nuclear magnetic resonance (NMR) spectroscopy data and their relative configuration was assigned based on nuclear Overhauser effect (NOE) and vicinal ¹H-NMR coupling data. The compounds where tested for biological activities; labindoles A (1) and B (2) exhibited significant activity against Hepatitis C Virus, 9H-carbazole (3), 3-chloro-9H-carbazole (4) and 4-hydroxymethyl-quinoline (5) showed antifungal activities. Moreover, compound3had weak to moderate antibacterial activities, while labindoles A (1) and B (2) were devoid of significant antifungal and antibacterial effects.
2018-03-21T15:35:12Z
2018-03-21T15:35:12Z
2018-02-28
Article
Six Heterocyclic Metabolites from the Myxobacterium Labilithrix luteola. 2018, 23 (3) Molecules
1420-3049
29495640
10.3390/molecules23030542
http://hdl.handle.net/10033/621329
Molecules (Basel, Switzerland)
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6213532020-01-08T14:12:14Zcom_10033_620589col_10033_620590
Molecular characteristics and successful management of a respiratory syncytial virus outbreak among pediatric patients with hemato-oncological disease.
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.
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.
2018-04-13T09:35:32Z
2018-04-13T09:35:32Z
2018
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6213692019-08-30T11:25:43Zcom_10033_620589col_10033_620590
CD81 Receptor Regions outside the Large Extracellular Loop Determine Hepatitis C Virus Entry into Hepatoma Cells.
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.
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.
2018-05-09T12:50:19Z
2018-05-09T12:50:19Z
2018
Article
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
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6214342019-08-30T11:34:19Zcom_10033_620589col_10033_620590
Glycomics and Proteomics Approaches to Investigate Early Adenovirus-Host Cell Interactions.
Lasswitz, Lisa
Chandra, Naresh
Arnberg, Niklas
Gerold, Gisa
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
adenovirus
glycomis
host cell interactions
proteomics
virus entry
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.
2018-08-02T08:12:33Z
2018-08-02T08:12:33Z
2018-06-22
Article
1089-8638
29746851
10.1016/j.jmb.2018.04.039
http://hdl.handle.net/10033/621434
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Journal of molecular biology
oai:repository.helmholtz-hzi.de:10033/6214832019-08-30T11:32:38Zcom_10033_620589col_10033_620590col_10033_620596
Pentagalloylglucose, a highly bioavailable polyphenolic compound present in Cortex moutan, efficiently blocks hepatitis C virus entry.
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.
Antivirals
Bioavailability
Cortex moutan
Entry inhibitor
Hepatitis C virus
Natural compounds
Penta-O-Galloyl-Glucose
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
2018-09-17T11:40:52Z
2018-09-17T11:40:52Z
2017-01-01
Article
1872-9096
28923507
10.1016/j.antiviral.2017.09.006
http://hdl.handle.net/10033/621483
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Antiviral research
oai:repository.helmholtz-hzi.de:10033/6214522019-08-30T11:28:48Zcom_10033_620589col_10033_620596
Environmental Stability and Infectivity of Hepatitis C Virus (HCV) in Different Human Body Fluids.
Pfaender, Stephanie
Helfritz, Fabian A
Siddharta, Anindya
Todt, Daniel
Behrendt, Patrick
Heyden, Julia
Riebesehl, Nina
Willmann, Wiebke
Steinmann, Joerg
Münch, Jan
Ciesek, Sandra
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
cerebrospinal fluid
contact lens solution
hepatitis C virus
infectivity
saliva
semen
tear
Hepatitis C virus (HCV) is a hepatotropic, blood-borne virus, but in up to one-third of infections of the transmission route remained unidentified. Viral genome copies of HCV have been identified in several body fluids, however, non-parental transmission upon exposure to contaminated body fluids seems to be rare. Several body fluids, e.g., tears and saliva, are renowned for their antimicrobial and antiviral properties, nevertheless, HCV stability has never been systematically analyzed in those fluids.
2018-08-27T08:26:48Z
2018-08-27T08:26:48Z
2018-01-01
Article
1664-302X
29636728
10.3389/fmicb.2018.00504
http://hdl.handle.net/10033/621452
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Frontiers in microbiology
oai:repository.helmholtz-hzi.de:10033/6214812019-08-30T11:29:14Zcom_10033_620589col_10033_620590
Hepatitis C virus enters liver cells using the CD81 receptor complex proteins calpain-5 and CBLB.
Bruening, Janina
Lasswitz, Lisa
Banse, Pia
Kahl, Sina
Marinach, Carine
Vondran, Florian W
Kaderali, Lars
Silvie, Olivier
Pietschmann, Thomas
Meissner, Felix
Gerold, Gisa
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.
2018-09-17T07:08:53Z
2018-09-17T07:08:53Z
2018-07-01
Article
1553-7374
30024968
10.1371/journal.ppat.1007111
http://hdl.handle.net/10033/621481
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
PLoS pathogens
oai:repository.helmholtz-hzi.de:10033/6214782019-08-30T11:27:14Zcom_10033_620589col_10033_620596
Virucidal efficacy of a sonicated hydrogen peroxide system (trophon EPR) following European and German test methods.
Becker, Britta
Bischoff, Birte
Brill, Florian H H
Steinmann, Eike
Steinmann, Jochen
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
disinfection
human papillomaviruses
trophon® EPR
ultrasound probes
virucidal efficacy
The virucidal efficacy of an automated ultrasound probe disinfector (trophon® EPR) was evaluated in a three step procedure according to European and German test methods. This system uses sonicated hydrogen peroxide mist (35%) at elevated temperature (50°C) in a closed chamber with control of all parameters within a 7 minute cycle.
Methods: In the first step of examination, the peroxide solution was tested in a quantitative suspension assay according to the Guideline of Deutsche Vereinigung zur Bekämpfung der Viruskrankheiten (DVV) e.V. and Robert Koch-Institute (RKI) and in parallel with the European Norm EN 14476 with all test viruses creating a virucidal claim.
In the second step, the virucidal efficacy of the hydrogen peroxide solution was evaluated in a hard surface carrier test according to the Guideline of DVV with adenovirus, murine norovirus and parvovirus simulating practical conditions.
Finally, the efficacy was evaluated by the automated system using stainless steel carriers inoculated with test virus and positioned at different levels inside the chamber.
2018-09-13T13:38:52Z
2018-09-13T13:38:52Z
2017-01-01
Article
2196-5226
28149707
10.3205/dgkh000287
http://hdl.handle.net/10033/621478
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
GMS hygiene and infection control
oai:repository.helmholtz-hzi.de:10033/6214862018-09-19T01:29:06Zcom_10033_620589col_10033_620596
Virucidal efficacy of a sonicated hydrogen peroxide system (trophon EPR) following European and German test methods.
Becker, Britta
Bischoff, Birte
Brill, Florian H H
Steinmann, Eike
Steinmann, Jochen
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
disinfection
human papillomaviruses
trophon® EPR
ultrasound probes
virucidal efficacy
The virucidal efficacy of an automated ultrasound probe disinfector (trophon® EPR) was evaluated in a three step procedure according to European and German test methods. This system uses sonicated hydrogen peroxide mist (35%) at elevated temperature (50°C) in a closed chamber with control of all parameters within a 7 minute cycle.
Methods: In the first step of examination, the peroxide solution was tested in a quantitative suspension assay according to the Guideline of Deutsche Vereinigung zur Bekämpfung der Viruskrankheiten (DVV) e.V. and Robert Koch-Institute (RKI) and in parallel with the European Norm EN 14476 with all test viruses creating a virucidal claim.
In the second step, the virucidal efficacy of the hydrogen peroxide solution was evaluated in a hard surface carrier test according to the Guideline of DVV with adenovirus, murine norovirus and parvovirus simulating practical conditions.
Finally, the efficacy was evaluated by the automated system using stainless steel carriers inoculated with test virus and positioned at different levels inside the chamber.
Results: A ≥4 log10 reduction of virus titre was demonstrated with all methods including carrier tests with murine norovirus, adenovirus, and parvovirus using the automated device.
Conclusion: The automated device is able to inactivate test viruses of German and European norms and can therefore claim efficacy against human pathogenic enveloped and non-enveloped viruses. This includes human papillomaviruses which form part of the complete virucidal claim.
2018-09-18T12:32:47Z
2018-09-18T12:32:47Z
2017-01-01
Article
2196-5226
28149707
10.3205/dgkh000287
http://hdl.handle.net/10033/621486
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
GMS hygiene and infection control
oai:repository.helmholtz-hzi.de:10033/6215132019-08-30T11:29:45Zcom_10033_620589col_10033_620596
Prevalence and characterization of azole-resistant Aspergillus fumigatus in patients with cystic fibrosis: a prospective multicentre study in Germany.
Seufert, R
Sedlacek, L
Kahl, B
Hogardt, M
Hamprecht, A
Haase, G
Gunzer, F
Haas, A
Grauling-Halama, S
MacKenzie, C R
Essig, A
Stehling, F
Sutharsan, S
Dittmer, S
Killengray, D
Schmidt, D
Eskandarian, N
Steinmann, E
Buer, J
Hagen, F
Meis, J F
Rath, P-M
Steinmann, J
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
Aspergillus fumigatus is the most prevalent filamentous fungus in the respiratory tract of patients with cystic fibrosis (CF). The aim of this prospective multicentre study was to investigate the prevalence of azole-resistant A. fumigatus (ARAF) in respiratory secretions from CF patients across Germany and to characterize ARAF isolates by phenotypic and molecular methods. Twelve tertiary care centres from Germany participated in the study. In total, 2888 A. fumigatus isolates from 961 CF patients were screened for ARAF by using azole-containing agar plates. Antifungal susceptibility testing of isolates was performed by broth microdilution according to EUCAST guidelines. Analysis of mutations mediating resistance was performed using PCR and sequencing of the cyp51A gene. Furthermore, genotyping by microsatellite PCR was performed. Of a total of 2888 A. fumigatus isolates, 101 isolates from 51 CF patients were found to be azole resistant (prevalence per patient 5.3%). The Essen centre had the highest prevalence (9.1%) followed by Munich (7.8%), Münster (6.0%) and Hannover (5.2%). Most ARAF isolates (n = 89) carried the TR34/L98H mutation followed by eight G54E/R, one TR46/Y121F/T289A and one F219S mutation. In two isolates no mutation was found. Genotyping results showed no major clustering. Forty-five percent of CF patients with ARAF had previously received azole therapy. This is the first multicentre study analysing the prevalence of ARAF isolates in German CF patients. Because of a resistance rate of up to 9%, susceptibility testing of A. fumigatus isolates from CF patients receiving antifungal treatment should be part of standard diagnostic work-up.
2018-10-11T07:11:20Z
2018-10-11T07:11:20Z
2018-08-01
Article
1460-2091
29684150
10.1093/jac/dky147
http://hdl.handle.net/10033/621513
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
The Journal of antimicrobial chemotherapy
oai:repository.helmholtz-hzi.de:10033/6215182019-08-30T11:29:45Zcom_10033_620589col_10033_620590col_10033_620596
The natural compound silvestrol inhibits hepatitis E virus (HEV) replication in vitro and in vivo.
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.
Antiviral activity
Hepatitis E virus (HEV)
Host target
Humanized mice
Replication
Silvestrol
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.
2018-10-12T10:49:42Z
2018-10-12T10:49:42Z
2018-09-01
Article
1872-9096
30036559
10.1016/j.antiviral.2018.07.010
http://hdl.handle.net/10033/621518
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Antiviral research
oai:repository.helmholtz-hzi.de:10033/6215492019-08-30T11:32:11Zcom_10033_620589col_10033_620590
The Small-Compound Inhibitor K22 Displays Broad Antiviral Activity against Different Members of the Family Flaviviridae and Offers Potential as a Panviral Inhibitor.
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.
Flaviviridae
K22
antiviral
flavivirus
hepacivirus
panviral inhibitor
pestivirus
The virus family
2018-11-09T10:16:10Z
2018-11-09T10:16:10Z
2018-11-01
Article
1098-6596
30181371
10.1128/AAC.01206-18
http://hdl.handle.net/10033/621549
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Antimicrobial agents and chemotherapy
oai:repository.helmholtz-hzi.de:10033/6216062019-08-30T11:31:43Zcom_10033_620589col_10033_620590
Kaposi's Sarcoma-Associated Herpesvirus Nonstructural Membrane Protein pK15 Recruits the Class II Phosphatidylinositol 3-Kinase PI3K-C2α To Activate Productive Viral Replication.
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.
KSHV-K15
PI3K-C2α
lytic replication
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.
2018-12-05T13:45:30Z
2018-12-05T13:45:30Z
2018-09-01
Article
1098-5514
29950425
10.1128/JVI.00544-18
http://hdl.handle.net/10033/621606
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Journal of virology
oai:repository.helmholtz-hzi.de:10033/6215522018-11-11T08:36:34Zcom_10033_620589col_10033_620595col_10033_620596
Susceptibility of Chikungunya Virus to Inactivation by Heat and Commercially and World Health Organization-Recommended Biocides.
Franz, Sergej
Friesland, Martina
Passos, Vânia
Todt, Daniel
Simmons, Graham
Goffinet, Christine
Steinmann, Eike
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Despite increasing clinical relevance of Chikungunya virus (CHIKV) infection, caused by a rapidly emerging pathogen, recommended guidelines for its inactivation do not exist. In this study, we investigated the susceptibility of CHIKV to inactivation by heat and commercially available hand, surface, and World Health Organization-recommended disinfectants to define CHIKV prevention protocols for healthcare systems.
2018-11-09T15:18:14Z
2018-11-09T15:18:14Z
2018-09-22
Article
1537-6613
29917109
10.1093/infdis/jiy359
http://hdl.handle.net/10033/621552
Attribution-NonCommercial-ShareAlike 3.0 United States
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
The Journal of infectious diseases
oai_dc///com_10033_620589/100