2024-03-29T09:26:42Zhttp://repository.helmholtz-hzi.de/oai/requestoai:repository.helmholtz-hzi.de:10033/1240272019-08-30T11:26:42Zcom_10033_311624com_10033_6839com_10033_311308col_10033_311625col_10033_620721
00925njm 22002777a 4500
dc
Solbak, Sara M
author
Reksten, Tove R
author
Wray, Victor
author
Bruns, Karsten
author
Horvli, Ole
author
Raae, Arnt J
author
Henklein, Petra
author
Henklein, Peter
author
Röder, Rene
author
Mitzner, David
author
Schubert, Ulrich
author
Fossen, Torgils
author
2010
Cyclophilin A (CypA) represents a potential target for antiretroviral therapy since inhibition of CypA suppresses human immunodeficiency virus type 1 (HIV-1) replication, although the mechanism through which CypA modulates HIV-1 infectivity still remains unclear. The interaction of HIV-1 viral protein R (Vpr) with the human peptidyl prolyl isomerase CypA is known to occur in vitro and in vivo. However, the nature of the interaction of CypA with Pro-35 of N-terminal Vpr has remained undefined.
The intriguing cyclophilin A-HIV-1 Vpr interaction: prolyl cis/trans isomerisation catalysis and specific binding. 2010, 10:31 BMC Struct. Biol.
1472-6807
20920334
10.1186/1472-6807-10-31
http://hdl.handle.net/10033/124027
BMC structural biology
The intriguing cyclophilin A-HIV-1 Vpr interaction: prolyl cis/trans isomerisation catalysis and specific binding.
oai:repository.helmholtz-hzi.de:10033/2369892019-08-30T11:25:43Zcom_10033_311624com_10033_6839com_10033_620636col_10033_311625col_10033_620638
00925njm 22002777a 4500
dc
Göhler, Anna-Katharina
author
Kökpinar, Öznur
author
Schmidt-Heck, Wolfgang
author
Geffers, Robert
author
Guthke, Reinhard
author
Rinas, Ursula
author
Schuster, Stefan
author
Jahreis, Knut
author
Kaleta, Christoph
author
2011
The pyruvate dehydrogenase regulator protein (PdhR) of Escherichia coli acts as a transcriptional regulator in a pyruvate dependent manner to control central metabolic fluxes. However, the complete PdhR regulon has not yet been uncovered. To achieve an extended understanding of its gene regulatory network, we combined large-scale network inference and experimental verification of results obtained by a systems biology approach.
More than just a metabolic regulator--elucidation and validation of new targets of PdhR in Escherichia coli. 2011, 5:197 BMC Syst Biol
1752-0509
22168595
10.1186/1752-0509-5-197
http://hdl.handle.net/10033/236989
BMC systems biology
More than just a metabolic regulator--elucidation and validation of new targets of PdhR in Escherichia coli.
oai:repository.helmholtz-hzi.de:10033/2970562019-08-30T11:33:05Zcom_10033_311624com_10033_6839com_10033_311308com_10033_338554col_10033_621787col_10033_311625col_10033_620777
00925njm 22002777a 4500
dc
Khetarpal, Niyati
author
Poddar, Ankur
author
Nemani, Satish K
author
Dhar, Nisha
author
Patil, Aravind
author
Negi, Priyanka
author
Perween, Ashiya
author
Viswanathan, Ramaswamy
author
Lünsdorf, Heinrich
author
Tyagi, Poornima
author
Raut, Rajendra
author
Arora, Upasana
author
Jain, Swatantra K
author
Rinas, Ursula
author
Swaminathan, Sathyamangalam
author
Khanna, Navin
author
2013
Dengue is today the most significant of arboviral diseases. Novel tools are necessary to effectively address the problem of dengue. Virus-like particles (VLP) offer a versatile nanoscale platform for developing tools with potential biomedical applications. From the perspective of a potentially useful dengue-specific tool, the dengue virus envelope protein domain III (EDIII), endowed with serotype-specificity, host receptor recognition and the capacity to elicit virus-neutralizing antibodies, is an attractive candidate.
Dengue-specific subviral nanoparticles: design, creation and characterization. 2013, 11 (1):15 J Nanobiotechnology
1477-3155
23706089
10.1186/1477-3155-11-15
http://hdl.handle.net/10033/297056
Journal of nanobiotechnology
Dengue-specific subviral nanoparticles: design, creation and characterization.
oai:repository.helmholtz-hzi.de:10033/3116262019-08-30T11:27:16Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Lang, Elke
author
Reichenbach, Hans
author
2013-11
Ten species of the order Myxococcales with validly published names are devoid of living type strains. Four species of the genus Chondromyces are represented by dead herbarium samples as the type material. For a species of the genus Melittangium and two species of the genus Polyangium, no physical type material was assigned at the time of validation of the names or later on. In accordance with rule 18f of the International Code of Nomenclature of Bacteria the following type strains are designated for these species: strain Cm a14(T) ( = DSM 14605(T) = JCM 12615(T)) as the type strain of Chondromyces apiculatus, strain Cm c5(T) ( = DSM 14714(T) = JCM 12616(T)) as the type strain of Chondromyces crocatus, strain Sy t2(T) ( = DSM 14631(T) = JCM 12617(T)) as the type strain of Chondromyces lanuginosus, strain Cm p51(T) ( = DSM 14607(T) = JCM 12618(T)) as the type strain of Chondromyces pediculatus, strain Me b8(T) ( = DSM 14713(T) = JCM 12633(T)) as the type strain of Melittangium boletus, strain Pl s12(T) ( = DSM 14670(T) = JCM 12637(T)) as the type strain of Polyangium sorediatum and strain Pl sm5(T) ( = DSM 14734(T) = JCM 12638(T)) as the type strain of Polyangium spumosum. Furthermore, the type strains given for three species of the genera Cystobacter and Polyangium had been kept at one university institute and have been lost according to our investigations. In accordance with Rule 18c of the Bacteriological Code, we propose the following neotype strains: strain Cb fe18 ( = DSM 14716 = JCM 12624) as the neotype strain of Cystobacter ferrugineus, strain Cb m2 ( = DSM 14751 = JCM 12627) as the neotype strain of Cystobacter minus and strain Pl fu5 ( = DSM 14668 = JCM 12636) as the neotype strain of Polyangium fumosum. The proposals of the strains are based on the descriptions and strain proposals given in the respective chapters of Bergey's Manual of Systematic Bacteriology (2005).
Designation of type strains for seven species of the order Myxococcales and proposal for neotype strains of Cystobacter ferrugineus, Cystobacter minus and Polyangium fumosum. 2013, 63 (Pt 11):4354-60 Int. J. Syst. Evol. Microbiol.
1466-5034
24187023
10.1099/ijs.0.056440-0
http://hdl.handle.net/10033/311626
International journal of systematic and evolutionary microbiology
Designation of type strains for seven species of the order Myxococcales and proposal for neotype strains of Cystobacter ferrugineus, Cystobacter minus and Polyangium fumosum.
oai:repository.helmholtz-hzi.de:10033/3464082019-08-30T11:31:23Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Esatbeyoglu, Tuba
author
Wray, Victor
author
Winterhalter, Peter
author
2015-07-15
The main procyanidins, including dimeric B2 and B5, trimeric C1, tetrameric and pentameric procyanidins, were isolated from unroasted cocoa beans (Theobroma cacao L.) using various techniques of countercurrent chromatography, such as high-speed countercurrent chromatography (HSCCC), low-speed rotary countercurrent chromatography (LSRCCC) and spiral-coil LSRCCC. Furthermore, dimeric procyanidins B1 and B7, which are not present naturally in the analysed cocoa beans, were obtained after semisynthesis of cocoa bean polymers with (+)-catechin as nucleophile and separated by countercurrent chromatography. In this way, the isolation of dimeric procyanidin B1 in considerable amounts (500mg, purity>97%) was possible in a single run. This is the first report concerning the isolation and semisynthesis of dimeric to pentameric procyanidins from T. cacao by countercurrent chromatography. Additionally, the chemical structures of tetrameric (cinnamtannin A2) and pentameric procyanidins (cinnamtannin A3) were elucidated on the basis of (1)H NMR spectroscopy. Interflavanoid linkage was determined by NOE-correlations, for the first time.
Isolation of dimeric, trimeric, tetrameric and pentameric procyanidins from unroasted cocoa beans (Theobroma cacao L.) using countercurrent chromatography. 2015, 179:278-89 Food Chem
0308-8146
25722166
10.1016/j.foodchem.2015.01.130
http://hdl.handle.net/10033/346408
Food chemistry
Isolation of dimeric, trimeric, tetrameric and pentameric procyanidins from unroasted cocoa beans (Theobroma cacao L.) using countercurrent chromatography.
oai:repository.helmholtz-hzi.de:10033/5588392019-08-30T11:29:17Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Rasch, Janine
author
Theuerkorn, Martin
author
Ünal, Can
author
Heinsohn, Natascha
author
Tran, Stefan
author
Fischer, Gunter
author
Weiwad, Matthias
author
Steinert, Michael
author
2015
Macrophage infectivity potentiator (Mip) and Mip-like proteins are virulence factors in a wide range of pathogens including Legionella pneumophila. These proteins belong to the FK506 binding protein (FKBP) family of peptidyl-prolyl-cis/trans-isomerases (PPIases). In L. pneumophila, the PPIase activity of Mip is required for invasion of macrophages, transmigration through an in vitro lung-epithelial barrier, and full virulence in the guinea pig infection model. Additionally, Mip is a moonlighting protein that binds to collagen IV in the extracellular matrix. Here, we describe the development and synthesis of cycloheximide derivatives with adamantyl moieties as novel FKBP ligands, and analyze their effect on the viability of L. pneumophila and other bacteria. All compounds efficiently inhibited PPIase activity of the prototypic human FKBP12 as well as Mip with IC50-values as low as 180 nM and 1.7 μM, respectively. Five of these derivatives inhibited the growth of L. pneumophila at concentrations of 30-40 μM, but exhibited no effect on other tested bacterial species indicating a specific spectrum of antibacterial activity. The derivatives carrying a 3,5-dimethyladamantan-1-[yl]acetamide substitution (MT_30.32), and a 3-ethyladamantan-1-[yl]acetamide substitution (MT_30.51) had the strongest effects in PPIase- and liquid growth assays. MT_30.32 and MT_30.51 were also inhibitory in macrophage infection studies without being cytotoxic. Accordingly, by applying a combinatorial approach, we were able to generate novel, hybrid inhibitors consisting of cycloheximide and adamantane, two known FKBP inhibitors that interact with different parts of the PPIase domain, respectively. Interestingly, despite the proven Mip-inhibitory activity, the viability of a Mip-deficient strain was affected to the same degree as its wild type. Hence, we also propose that cycloheximide derivatives with adamantyl moieties are potent PPIase inhibitors with multiple targets in L. pneumophila.
Novel Cycloheximide Derivatives Targeting the Moonlighting Protein Mip Exhibit Specific Antimicrobial Activity Against Legionella pneumophila. 2015, 3:41 Front Bioeng Biotechnol
2296-4185
25870856
10.3389/fbioe.2015.00041
http://hdl.handle.net/10033/558839
Frontiers in bioengineering and biotechnology
Novel Cycloheximide Derivatives Targeting the Moonlighting Protein Mip Exhibit Specific Antimicrobial Activity Against Legionella pneumophila.
oai:repository.helmholtz-hzi.de:10033/5588272019-08-30T11:27:16Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Xiong, Qiuhong
author
Ünal, Can
author
Matthias, Jan
author
Steinert, Michael
author
Eichinger, Ludwig
author
2015-04
Macroautophagy is a highly conserved intracellular bulk degradation system of all eukaryotic cells. It is governed by a large number of autophagy proteins (ATGs) and is crucial for many cellular processes. Here, we describe the phenotypes of Dictyostelium discoideum ATG16(-) and ATG9(-)/16(-) cells and compare them to the previously reported ATG9(-) mutant. ATG16 deficiency caused an increase in the expression of several core autophagy genes, among them atg9 and the two atg8 paralogues. The single and double ATG9 and ATG16 knock-out mutants had complex phenotypes and displayed severe and comparable defects in pinocytosis and phagocytosis. Uptake of Legionella pneumophila was reduced. In addition, ATG9(-) and ATG16(-) cells had dramatic defects in autophagy, development and proteasomal activity which were much more severe in the ATG9(-)/16(-) double mutant. Mutant cells showed an increase in poly-ubiquitinated proteins and contained large ubiquitin-positive protein aggregates which partially co-localized with ATG16-GFP in ATG9(-)/16(-) cells. The more severe autophagic, developmental and proteasomal phenotypes of ATG9(-)/16(-) cells imply that ATG9 and ATG16 probably function in parallel in autophagy and have in addition autophagy-independent functions in further cellular processes.
The phenotypes of ATG9, ATG16 and ATG9/16 knock-out mutants imply autophagy-dependent and -independent functions. 2015, 5 (4):150008 Open Biol
2046-2441
25878144
10.1098/rsob.150008
http://hdl.handle.net/10033/558827
Open biology
The phenotypes of ATG9, ATG16 and ATG9/16 knock-out mutants imply autophagy-dependent and -independent functions.
oai:repository.helmholtz-hzi.de:10033/5840772019-08-30T11:34:48Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Ünal, Can M
author
Steinert, Michael
author
2015-10
FK506-binding proteins (FKBPs) contain a domain with peptidyl-prolyl-cis/trans-isomerase (PPIase) activity and bind the immunosuppressive drugs FK506 and rapamycin. FKBPs belong to the immunophilin family and are found in eukaryotes and bacteria.
FKBPs in bacterial infections. 2015, 1850 (10):2096-102 Biochim. Biophys. Acta
0006-3002
25529296
10.1016/j.bbagen.2014.12.018
http://hdl.handle.net/10033/584077
Biochimica et biophysica acta
FKBPs in bacterial infections.
oai:repository.helmholtz-hzi.de:10033/6138722019-08-30T11:33:30Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Rueda, Fabián
author
Céspedes, María Virtudes
author
Sánchez-Chardi, Alejandro
author
Seras-Franzoso, Joaquin
author
Pesarrodona, Mireia
author
Ferrer-Miralles, Neus
author
Vázquez, Esther
author
Rinas, Ursula
author
Unzueta, Ugutz
author
Mamat, Uwe
author
Mangues, Ramón
author
García-Fruitós, Elena
author
Villaverde, Antonio
author
2016
Production of recombinant drugs in process-friendly endotoxin-free bacterial factories targets to a lessened complexity of the purification process combined with minimized biological hazards during product application. The development of nanostructured recombinant materials in innovative nanomedical activities expands such a need beyond plain functional polypeptides to complex protein assemblies. While Escherichia coli has been recently modified for the production of endotoxin-free proteins, no data has been so far recorded regarding how the system performs in the fabrication of smart nanostructured materials.
Structural and functional features of self-assembling protein nanoparticles produced in endotoxin-free Escherichia coli. 2016, 15 (1):59 Microb. Cell Fact.
1475-2859
27059706
10.1186/s12934-016-0457-z
http://hdl.handle.net/10033/613872
Microbial cell factories
Structural and functional features of self-assembling protein nanoparticles produced in endotoxin-free Escherichia coli.
oai:repository.helmholtz-hzi.de:10033/6208562019-08-30T11:31:23Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Schmid, Ursula
author
Stenzel, Werner
author
Koschel, Josephin
author
Raptaki, Maria
author
Wang, Xu
author
Naumann, Michael
author
Matuschewski, Kai
author
Schlüter, Dirk
author
Nishanth, Gopala
author
2017
Cerebral malaria is a severe complication of human malaria and may lead to death of Plasmodium falciparum-infected individuals. Cerebral malaria is associated with sequestration of parasitized red blood cells within the cerebral microvasculature resulting in damage of the blood-brain barrier and brain pathology. Although CD8(+) T cells have been implicated in the development of murine experimental cerebral malaria (ECM), several other studies have shown that CD8(+) T cells confer protection against blood-stage infections. Since the role of host deubiquitinating enzymes (DUBs) in malaria is yet unknown, we investigated how the DUB cylindromatosis (CYLD), an important inhibitor of several cellular signaling pathways, influences the outcome of ECM. Upon infection with Plasmodium berghei ANKA (PbA) sporozoites or PbA-infected red blood cells, at least 90% of Cyld(-/-) mice survived the infection, whereas all congenic C57BL/6 mice displayed signatures of ECM, impaired parasite control, and disruption of the blood-brain barrier integrity. Cyld deficiency prevented brain pathology, including hemorrhagic lesions, enhanced activation of astrocytes and microglia, infiltration of CD8(+) T cells, and apoptosis of endothelial cells. Furthermore, PbA-specific CD8(+) T cell responses were augmented in the blood of Cyld(-/-) mice with increased production of interferon-γ and granzyme B and elevated activation of protein kinase C-θ and nuclear factor "kappa light-chain enhancer" of activated B cells. Importantly, accumulation of CD8(+) T cells in the brain of Cyld(-/-) mice was significantly reduced compared to C57BL/6 mice. Bone marrow chimera experiments showed that the absence of ECM signatures in infected Cyld(-/-) mice could be attributed to hematopoietic and radioresistant parenchymal cells, most likely endothelial cells that did not undergo apoptosis. Together, we were able to show that host deubiqutinating enzymes play an important role in ECM and that CYLD promotes ECM supporting it as a potential therapeutic target for adjunct therapy to prevent cerebral complications of severe malaria.
The Deubiquitinating Enzyme Cylindromatosis Dampens CD8(+) T Cell Responses and Is a Critical Factor for Experimental Cerebral Malaria and Blood-Brain Barrier Damage. 2017, 8:27 Front Immunol
28203236
10.3389/fimmu.2017.00027
http://hdl.handle.net/10033/620856
Frontiers in immunology
The Deubiquitinating Enzyme Cylindromatosis Dampens CD8(+) T Cell Responses and Is a Critical Factor for Experimental Cerebral Malaria and Blood-Brain Barrier Damage.
oai:repository.helmholtz-hzi.de:10033/6208942019-08-30T11:37:23Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Hoppe, Julia
author
Ünal, Can M
author
Thiem, Stefanie
author
Grimpe, Louisa
author
Goldmann, Torsten
author
Gaßler, Nikolaus
author
Richter, Matthias
author
Shevchuk, Olga
author
Steinert, Michael
author
2017
Legionnaires' disease is an acute fibrinopurulent pneumonia. During infection Legionella pneumophila adheres to the alveolar lining and replicates intracellularly within recruited macrophages. Here we provide a sequence and domain composition analysis of the L. pneumophila PilY1 protein, which has a high homology to PilY1 of Pseudomonas aeruginosa. PilY1 proteins of both pathogens contain a von Willebrand factor A (vWFa) and a C-terminal PilY domain. Using cellular fractionation, we assigned the L. pneumophila PilY1 as an outer membrane protein that is only expressed during the transmissive stationary growth phase. PilY1 contributes to infection of human lung tissue explants (HLTEs). A detailed analysis using THP-1 macrophages and A549 lung epithelial cells revealed that this contribution is due to multiple effects depending on host cell type. Deletion of PilY1 resulted in a lower replication rate in THP-1 macrophages but not in A549 cells. Further on, adhesion to THP-1 macrophages and A549 epithelial cells was decreased. Additionally, the invasion into non-phagocytic A549 epithelial cells was drastically reduced when PilY1 was absent. Complementation variants of a PilY1-negative mutant revealed that the C-terminal PilY domain is essential for restoring the wild type phenotype in adhesion, while the putatively mechanosensitive vWFa domain facilitates invasion into non-phagocytic cells. Since PilY1 also promotes twitching motility of L. pneumophila, we discuss the putative contribution of this newly described virulence factor for bacterial dissemination within infected lung tissue.
PilY1 Promotes Legionella pneumophila Infection of Human Lung Tissue Explants and Contributes to Bacterial Adhesion, Host Cell Invasion, and Twitching Motility. 2017, 7:63 Front Cell Infect Microbiol
2235-2988
28326293
10.3389/fcimb.2017.00063
http://hdl.handle.net/10033/620894
Frontiers in cellular and infection microbiology
PilY1 Promotes Legionella pneumophila Infection of Human Lung Tissue Explants and Contributes to Bacterial Adhesion, Host Cell Invasion, and Twitching Motility.
oai:repository.helmholtz-hzi.de:10033/6209302019-08-30T11:29:47Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Just, Sissy
author
Nishanth, Gopala
author
Buchbinder, Jörn H
author
Wang, Xu
author
Naumann, Michael
author
Lavrik, Inna
author
Schlüter, Dirk
author
2016-12-22
The ubiquitin-modifying enzyme A20, an important negative feedback regulator of NF-κB, impairs the expansion of tumor-specific CD8(+) T cells but augments the proliferation of autoimmune CD4(+) T cells. To study the T cell-specific function of A20 in bacterial infection, we infected T cell-specific A20 knockout (CD4-Cre A20(fl/fl)) and control mice with Listeria monocytogenes. A20-deficient pathogen-specific CD8(+) T cells expanded stronger resulting in improved pathogen control at day 7 p.i. Imaging flow cytometry revealed that A20-deficient Listeria-specific CD8(+) T cells underwent increased apoptosis and necroptosis resulting in reduced numbers of memory CD8(+) T cells. In contrast, the primary CD4(+) T cell response was A20-independent. Upon secondary infection, the increase and function of pathogen-specific CD8(+) T cells, as well as pathogen control were significantly impaired in CD4-Cre A20(fl/fl) mice. In vitro, apoptosis and necroptosis of Listeria-specific A20-deficient CD8(+) T cells were strongly induced as demonstrated by increased caspase-3/7 activity, RIPK1/RIPK3 complex formation and more morphologically apoptotic and necroptotic CD8(+) T cells. In vitro, A20 limited CD95L and TNF-induced caspase3/7 activation. In conclusion, T cell-specific A20 limited the expansion but reduced apoptosis and necroptosis of Listeria-specific CD8(+) T cells, resulting in an impaired pathogen control in primary but improved clearance in secondary infection.
A20 Curtails Primary but Augments Secondary CD8(+) T Cell Responses in Intracellular Bacterial Infection. 2016, 6:39796 Sci Rep
2045-2322
28004776
10.1038/srep39796
http://hdl.handle.net/10033/620930
Scientific reports
A20 Curtails Primary but Augments Secondary CD8(+) T Cell Responses in Intracellular Bacterial Infection.
oai:repository.helmholtz-hzi.de:10033/6209722019-08-30T11:32:16Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Muruganandam, Gopinath
author
Raasakka, Arne
author
Myllykoski, Matti
author
Kursula, Inari
author
Kursula, Petri
author
2017-05-16
Eukaryotic tRNA splicing is an essential process in the transformation of a primary tRNA transcript into a mature functional tRNA molecule. 5'-phosphate ligation involves two steps: a healing reaction catalyzed by polynucleotide kinase (PNK) in association with cyclic phosphodiesterase (CPDase), and a sealing reaction catalyzed by an RNA ligase. The enzymes that catalyze tRNA healing in yeast and higher eukaryotes are homologous to the members of the 2H phosphoesterase superfamily, in particular to the vertebrate myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase).
Structural similarities and functional differences clarify evolutionary relationships between tRNA healing enzymes and the myelin enzyme CNPase. 2017, 18 (1):7 BMC Biochem.
1471-2091
28511668
10.1186/s12858-017-0084-2
http://hdl.handle.net/10033/620972
BMC biochemistry
Structural similarities and functional differences clarify evolutionary relationships between tRNA healing enzymes and the myelin enzyme CNPase.
oai:repository.helmholtz-hzi.de:10033/6209802019-08-30T11:33:29Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Lim, Michelle C C
author
Maubach, Gunter
author
Sokolova, Olga
author
Feige, Michael H
author
Diezko, Rolf
author
Buchbinder, Jörn
author
Backert, Steffen
author
Schlüter, Dirk
author
Lavrik, Inna N
author
Naumann, Michael
author
2017-06-02
The human pathogen Helicobacter pylori infects more than half of the world's population and is a paradigm for persistent yet asymptomatic infection but increases the risk for chronic gastritis and gastric adenocarcinoma. For successful colonization, H. pylori needs to subvert the host cell death response, which serves to confine pathogen infection by killing infected cells and preventing malignant transformation. Infection of gastric epithelial cells by H. pylori provokes direct and fast activation of the proinflammatory and survival factor NF-κB, which regulates target genes, such as CXCL8, BIRC3 and TNFAIP3. However, it is not known how H. pylori exploits NF-κB activation and suppresses the inflammatory response and host apoptotic cell death, in order to avert the innate immune response and avoid cell loss, and thereby enhance colonization to establish long-term infection. Here we assign for the first time that H. pylori and also Campylobacter jejuni-induced ubiquitin-editing enzyme A20 bifunctionally terminates NF-κB activity and negatively regulates apoptotic cell death. Mechanistically, we show that the deubiquitinylase activity of A20 counteracts cullin3-mediated K63-linked ubiquitinylation of procaspase-8, therefore restricting the activity of caspase-8. Interestingly, another inducible NF-κB target gene, the scaffold protein p62, ameliorates the interaction of A20 with procaspase-8. In conclusion, pathogen-induced de novo synthesis of A20 regulates the shut-off of the survival factor NF-κB but, on the other hand, also impedes caspase-8-dependent apoptotic cell death so as to promote the persistence of pathogens.Cell Death and Differentiation advance online publication, 2 June 2017; doi:10.1038/cdd.2017.89.
Pathogen-induced ubiquitin-editing enzyme A20 bifunctionally shuts off NF-κB and caspase-8-dependent apoptotic cell death. 2017 Cell Death Differ.
1476-5403
28574503
10.1038/cdd.2017.89
http://hdl.handle.net/10033/620980
Cell death and differentiation
Pathogen-induced ubiquitin-editing enzyme A20 bifunctionally shuts off NF-κB and caspase-8-dependent apoptotic cell death.
oai:repository.helmholtz-hzi.de:10033/6209842019-08-30T11:31:23Zcom_10033_311624com_10033_6839com_10033_620626com_10033_311308com_10033_338554col_10033_621787col_10033_311625col_10033_620721col_10033_620629
00925njm 22002777a 4500
dc
Rahim, Muhammad Imran
author
Babbar, Anshu
author
Lienenklaus, Stefan
author
Pils, Marina
author
Rohde, M
author
2017-06-01
Biomaterial-associated Pseudomonas aeruginosa biofilm infections constitute cascade of host immune reactions ultimately leading towards implant failure. Due to lack of relevant in vivo biofilm models, majority of the studies report host immune responses against free living or planktonic bacteria while bacteria in clinical situations live more frequently as biofilm communities than as single cells. Present study investigated host immune responses against biomaterial-associated P. aeruginosa biofilms in a clinically relevant mouse model. Previously, we reported metallic magnesium, a prospective biodegradable implant, to be permissive for bacterial biofilms in vivo even though it exhibits antibacterial properties in vitro. Therefore, magnesium was employed as biomaterial to investigate in vivo biofilm formation and associated host immune responses by using two P. aeruginosa strains and two mouse strains. P. aeruginosa formed biofilms on subcutaneously implanted magnesium discs. Non-invasive in vivo imaging indicated transient inflammatory responses at control sites whereas robust prolonged interferon-β (IFN-β) expression was observed from biofilms in a transgenic animal reporter. Further, immunohistology and electron microscopic results showed that bacterial biofilms were located in two dimensions immediately on the implant surface and at a short distance in the adjacent tissue. These biofilms were surrounded by inflammatory cells (mainly polymorphonuclear cells) as compared to controls. Interestingly, even though the number of live bacteria in various organs remained below detectable levels, splenomegaly indicated systemic inflammatory processes. Overall, these findings confirmed the resistance of biofilm infections in vivo to potentially antibacterial properties of magnesium degradation products. In vivo imaging and histology indicated the induction of both, local and systemic host inflammatory responses against P. aeruginosa biofilms. Even though the innate host immune defenses could not eliminate the local infection for up to two weeks, there was no apparent systemic bacteremia and all animals investigated survived the infection.
Degradable magnesium implant-associated infections by bacterial biofilms induce robust localized and systemic inflammatory reactions in a mouse model. 2017 Biomed Mater
1748-605X
28569671
10.1088/1748-605X/aa7667
http://hdl.handle.net/10033/620984
Biomedical materials (Bristol, England)
Degradable magnesium implant-associated infections by bacterial biofilms induce robust localized and systemic inflammatory reactions in a mouse model.
oai:repository.helmholtz-hzi.de:10033/6210242019-08-30T11:35:10Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Debarry, Jennifer
author
Heinz, D
author
Manns, M P
author
2017-07
[Individualized infection medicine : Challenges and opportunities]. 2017, 58 (7):647-649 Internist (Berl)
1432-1289
28589213
10.1007/s00108-017-0267-3
http://hdl.handle.net/10033/621024
Der Internist
[Individualized infection medicine : Challenges and opportunities].
oai:repository.helmholtz-hzi.de:10033/6210342019-08-30T11:36:32Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Kuhn, Philipp
author
Thiem, Stefanie
author
Steinert, Michael
author
Purvis, Duncan
author
Lugmayr, Veronika
author
Treutlein, Ulrich
author
Plobner, Lutz
author
Leiser, Robert-Matthias
author
Hust, Michael
author
Dübel, Stefan
author
2017-07-19
Legionella are Gram-negative bacteria that are ubiquitously present in natural and man-made water reservoirs. When humans inhale aerosolized water contaminated with Legionella, alveolar macrophages can be infected, which may lead to a life-threatening pneumonia called Legionnaires' disease. Due to the universal distribution of Legionella in water and their potential threat to human health, the Legionella concentration in water for human use must be strictly monitored, which is difficult since the standard detection still relies on lengthy cultivation and analysis of bacterial morphology. In this study, an antibody against L. pneumophila has been generated from the naïve human HAL antibody libraries by phage-display for the first time. The panning was performed on whole bacterial cells in order to select antibodies that bind specifically to the cell surface of untreated Legionella. The bacterial cell wall component lipopolysaccharide (LPS) was identified as the target structure. Specific binding to the important pathogenic L. pneumophila strains Corby, Philadelphia-1 and Knoxville was observed, while no binding was detected to seven members of the families Enterobacteriaceae, Pseudomonadaceae or Clostridiaceae. Production of this antibody in the recombinant scFv-Fc format using either a murine or a human Fc part allowed the set-up of a sandwich-ELISA for detection of Legionella cells. The scFv-Fc construct proved to be very stable, even when stored for several weeks at elevated temperatures. A sensitivity limit of 4,000 cells was achieved. The scFv-Fc antibody pair was integrated on a biosensor, demonstrating the specific and fast detection of L. pneumophila on a portable device. With this system, 10,000 Legionella cells were detected within 35 min. Combined with a water filtration/concentration system, this antibody may be developed into a promising reagent for rapid on-site Legionella monitoring.
Human Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity. 2017, 26 (1):29-38 Hum Antibodies
1875-869X
28582852
10.3233/HAB-170318
http://hdl.handle.net/10033/621034
Human antibodies
Human Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity.
oai:repository.helmholtz-hzi.de:10033/6210512019-08-30T11:34:46Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Manns, Michael P
author
Huber, Petra
author
Jaeckel, Elmar
author
2017-08-09
28427110
10.1055/s-0043-101946
http://hdl.handle.net/10033/621051
Sonderforschungsbereich SFB 738: Optimierung konventioneller und innovativer Transplantate
oai:repository.helmholtz-hzi.de:10033/6210592019-08-30T11:34:22Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Link, Alexander
author
Langner, Cosima
author
Schirrmeister, Wiebke
author
Habendorf, Wiebke
author
Weigt, Jochen
author
Venerito, Marino
author
Tammer, Ina
author
Schlüter, Dirk
author
Schlaermann, Philipp
author
Meyer, Thomas F
author
Wex, Thomas
author
Malfertheiner, Peter
author
2017-07-14
To evaluate the frequency of Helicobacter pylori (H. pylori) CagA antibodies in H. pylori infected subjects and to identify potential histopathological and bacterial factors related to H. pylori CagA-immune response.
Helicobacter pylori vacA genotype is a predominant determinant of immune response to Helicobacter pylori CagA. 2017, 23 (26):4712-4723 World J. Gastroenterol.
2219-2840
28765692
10.3748/wjg.v23.i26.4712
http://hdl.handle.net/10033/621059
World journal of gastroenterology
Helicobacter pylori vacA genotype is a predominant determinant of immune response to Helicobacter pylori CagA.
oai:repository.helmholtz-hzi.de:10033/6210632019-08-30T11:36:05Zcom_10033_311624com_10033_6839com_10033_620636col_10033_311625col_10033_620638
00925njm 22002777a 4500
dc
Swierzy, Izabela J
author
Händel, Ulrike
author
Kaever, Alexander
author
Jarek, Michael
author
Scharfe, Maren
author
Schlüter, Dirk
author
Lüder, Carsten G K
author
2017-08-03
The apicomplexan parasite Toxoplasma gondii infects various cell types in avian and mammalian hosts including humans. Infection of immunocompetent hosts is mostly asymptomatic or benign, but leads to development of largely dormant bradyzoites that persist predominantly within neurons and muscle cells. Here we have analyzed the impact of the host cell type on the co-transcriptomes of host and parasite using high-throughput RNA sequencing. Murine cortical neurons and astrocytes, skeletal muscle cells (SkMCs) and fibroblasts differed by more than 16,200 differentially expressed genes (DEGs) before and after infection with T. gondii. However, only a few hundred of them were regulated by infection and these largely diverged in neurons, SkMCs, astrocytes and fibroblasts indicating host cell type-specific transcriptional responses after infection. The heterogeneous transcriptomes of host cells before and during infection coincided with ~5,400 DEGs in T. gondii residing in different cell types. Finally, we identified gene clusters in both T. gondii and its host, which correlated with the predominant parasite persistence in neurons or SkMCs as compared to astrocytes or fibroblasts. Thus, heterogeneous expression profiles of different host cell types and the parasites' ability to adapting to them may govern the parasite-host cell interaction during toxoplasmosis.
Divergent co-transcriptomes of different host cells infected with Toxoplasma gondii reveal cell type-specific host-parasite interactions. 2017, 7 (1):7229 Sci Rep
2045-2322
28775382
10.1038/s41598-017-07838-w
http://hdl.handle.net/10033/621063
Scientific reports
Divergent co-transcriptomes of different host cells infected with Toxoplasma gondii reveal cell type-specific host-parasite interactions.
oai:repository.helmholtz-hzi.de:10033/6210682019-08-30T11:27:16Zcom_10033_311624com_10033_6839com_10033_620652col_10033_620672col_10033_311625
00925njm 22002777a 4500
dc
Bhushal, Sudeep
author
Wolfsmüller, Markus
author
Selvakumar, Tharini A
author
Kemper, Lucas
author
Wirth, Dagmar
author
Hornef, Mathias W
author
Hauser, Hansjörg
author
Köster, Mario
author
2017
Type I and type III interferons (IFNs) are crucial components of the first-line antiviral host response. While specific receptors for both IFN types exist, intracellular signaling shares the same Jak-STAT pathway. Due to its receptor expression, IFN-λ responsiveness is restricted mainly to epithelial cells. Here, we display IFN-stimulated gene induction at the single cell level to comparatively analyze the activities of both IFN types in intestinal epithelial cells and mini-gut organoids. Initially, we noticed that the response to both types of IFNs at low concentrations is based on a single cell decision-making determining the total cell intrinsic antiviral activity. We identified histone deacetylase (HDAC) activity as a crucial restriction factor controlling the cell frequency of IFN-stimulated gene (ISG) induction upon IFN-λ but not IFN-β stimulation. Consistently, HDAC blockade confers antiviral activity to an elsewise non-responding subpopulation. Second, in contrast to the type I IFN system, polarization of intestinal epithelial cells strongly enhances their ability to respond to IFN-λ signaling and raises the kinetics of gene induction. Finally, we show that ISG induction in mini-gut organoids by low amounts of IFN is characterized by a scattered heterogeneous responsiveness of the epithelial cells and HDAC activity fine-tunes exclusively IFN-λ activity. This study provides a comprehensive description of the differential response to type I and type III IFNs and demonstrates that cell polarization in gut epithelial cells specifically increases IFN-λ activity.
Cell Polarization and Epigenetic Status Shape the Heterogeneous Response to Type III Interferons in Intestinal Epithelial Cells. 2017, 8:671 Front Immunol
1664-3224
28659914
10.3389/fimmu.2017.00671
http://hdl.handle.net/10033/621068
Frontiers in immunology
Cell Polarization and Epigenetic Status Shape the Heterogeneous Response to Type III Interferons in Intestinal Epithelial Cells.
oai:repository.helmholtz-hzi.de:10033/6211072019-08-30T11:27:16Zcom_10033_311624com_10033_6839com_10033_311308col_10033_311625col_10033_559591
00925njm 22002777a 4500
dc
Dolan, Stephen K
author
Bock, Tobias
author
Hering, Vanessa
author
Owens, Rebecca A
author
Jones, Gary W
author
Blankenfeldt, Wulf
author
Doyle, Sean
author
2017-02
Gliotoxin is an epipolythiodioxopiperazine (ETP) class toxin, contains a disulfide bridge that mediates its toxic effects via redox cycling and is produced by the opportunistic fungal pathogen Aspergillus fumigatus Self-resistance against gliotoxin is effected by the gliotoxin oxidase GliT, and attenuation of gliotoxin biosynthesis is catalysed by gliotoxin S-methyltransferase GtmA. Here we describe the X-ray crystal structures of GtmA-apo (1.66 Å), GtmA complexed to S-adenosylhomocysteine (1.33 Å) and GtmA complexed to S-adenosylmethionine (2.28 Å), providing mechanistic insights into this important biotransformation. We further reveal that simultaneous elimination of the ability of A. fumigatus to dissipate highly reactive dithiol gliotoxin, via deletion of GliT and GtmA, results in the most significant hypersensitivity to exogenous gliotoxin observed to date. Indeed, quantitative proteomic analysis of ΔgliT::ΔgtmA reveals an uncontrolled over-activation of the gli-cluster upon gliotoxin exposure. The data presented herein reveal, for the first time, the extreme risk associated with intracellular dithiol gliotoxin biosynthesis-in the absence of an efficient dismutation capacity. Significantly, a previously concealed protective role for GtmA and functionality of ETP bis-thiomethylation as an ancestral protection strategy against dithiol compounds is now evident.
Structural, mechanistic and functional insight into gliotoxin bis-thiomethylation in Aspergillus fumigatus. 2017, 7 (2) Open Biol
2046-2441
28179499
10.1098/rsob.160292
http://hdl.handle.net/10033/621107
Open biology
Structural, mechanistic and functional insight into gliotoxin bis-thiomethylation in Aspergillus fumigatus.
oai:repository.helmholtz-hzi.de:10033/6211202019-08-30T11:34:22Zcom_10033_311624com_10033_6839com_10033_620652col_10033_620672col_10033_311625
00925njm 22002777a 4500
dc
Cebula, Marcin
author
Riehn, Mathias
author
Hillebrand, Upneet
author
Kratzer, Ramona F
author
Kreppel, Florian
author
Koutsoumpli, Georgia
author
Daemen, Toos
author
Hauser, Hansjoerg
author
Wirth, Dagmar
author
2017-07-14
Immune defense against hepatotropic viruses such as hepatitis B (HBV) and hepatitis C (HCV) poses a major challenge for therapeutic approaches. Intrahepatic cytotoxic CD8 T cells that are crucial for an immune response against these viruses often become exhausted resulting in chronic infection. We elucidated the T cell response upon therapeutic vaccination in inducible transgenic mouse models in which variable percentages of antigen-expressing hepatocytes can be adjusted, providing mosaic antigen distribution and reflecting the varying viral antigen loads observed in patients. Vaccination-induced endogenous CD8 T cells could eliminate low antigen loads in liver but were functionally impaired if confronted with elevated antigen loads. Strikingly, only by conditioning the liver environment with TLR9 ligand prior and early after peripheral vaccination, successful immunization against high intrahepatic antigen density with its elimination was achieved. Moreover, TLR9 immunomodulation was also indispensable for functional memory recall after high frequency antigen challenge. Together, the results indicate that TLR9-mediated conditioning of liver environment during therapeutic vaccination or antigen reoccurrence is crucial for an efficacious intrahepatic T cell response.
TLR9-Mediated Conditioning of Liver Environment Is Essential for Successful Intrahepatic Immunotherapy and Effective Memory Recall. 2017 Mol. Ther.
1525-0024
28716576
10.1016/j.ymthe.2017.06.018
http://hdl.handle.net/10033/621120
Molecular therapy : the journal of the American Society of Gene Therapy
TLR9-Mediated Conditioning of Liver Environment Is Essential for Successful Intrahepatic Immunotherapy and Effective Memory Recall.
oai:repository.helmholtz-hzi.de:10033/6211282019-08-30T11:34:48Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Nonnenmacher, Yannic
author
Palorini, Roberta
author
d'Herouël, Aymeric Fouquier
author
Krämer, Lisa
author
Neumann-Schaal, Meina
author
Chiaradonna, Ferdinando
author
Skupin, Alexander
author
Wegner, Andre
author
Hiller, Karsten
author
2017-09
To date, it is well-established that mitochondrial dysfunction does not only play a vital role in cancer but also in other pathological conditions such as neurodegenerative diseases and inflammation. An important tool for the analysis of cellular metabolism is the application of stable isotope labeled substrates, which allow for the tracing of atoms throughout metabolic networks. While such analyses yield very detailed information about intracellular fluxes, the determination of compartment specific fluxes is far more challenging. Most approaches for the deconvolution of compartmented metabolism use computational models whereas experimental methods are rare. Here, we developed an experimental setup based on selective permeabilization of the cytosolic membrane that allows for the administration of stable isotope labeled substrates directly to mitochondria. We demonstrate how this approach can be used to infer metabolic changes in mitochondria induced by either chemical or genetic perturbations and give an outlook on its potential applications.
Analysis of mitochondrial metabolism in situ: Combining stable isotope labeling with selective permeabilization. 2017, 43 (Pt B):147-155 Metab. Eng.
1096-7184
27988388
10.1016/j.ymben.2016.12.005
http://hdl.handle.net/10033/621128
Metabolic engineering
Analysis of mitochondrial metabolism in situ: Combining stable isotope labeling with selective permeabilization.
oai:repository.helmholtz-hzi.de:10033/6211302019-08-30T11:28:51Zcom_10033_311624com_10033_6839com_10033_620652col_10033_620672col_10033_311625
00925njm 22002777a 4500
dc
Gödecke, Natascha
author
Zha, Lisha
author
Spencer, Shawal
author
Behme, Sara
author
Riemer, Pamela
author
Rehli, Michael
author
Hauser, Hansjörg
author
Wirth, Dagmar
author
2017-09-19
Faithful expression of transgenes in cell cultures and mice is often challenged by locus dependent epigenetic silencing. We investigated silencing of Tet-controlled expression cassettes within the mouse ROSA26 locus. We observed pronounced DNA methylation of the Tet promoter concomitant with loss of expression in mES cells as well as in differentiated cells and transgenic animals. Strikingly, the ROSA26 promoter remains active and methylation free indicating that this silencing mechanism specifically affects the transgene, but does not spread to the host's chromosomal neighborhood. To reactivate Tet cassettes a synthetic fusion protein was constructed and expressed in silenced cells. This protein includes the enzymatic domains of ten eleven translocation methylcytosine dioxygenase 1 (TET-1) as well as the Tet repressor DNA binding domain. Expression of the synthetic fusion protein and Doxycycline treatment allowed targeted demethylation of the Tet promoter in the ROSA26 locus and in another genomic site, rescuing transgene expression in cells and transgenic mice. Thus, inducible, reversible and site-specific epigenetic modulation is a promising strategy for reactivation of silenced transgene expression, independent of the integration site.
Controlled re-activation of epigenetically silenced Tet promoter-driven transgene expression by targeted demethylation. 2017, 45 (16):e147 Nucleic Acids Res.
1362-4962
28934472
10.1093/nar/gkx601
http://hdl.handle.net/10033/621130
Nucleic acids research
Controlled re-activation of epigenetically silenced Tet promoter-driven transgene expression by targeted demethylation.
oai:repository.helmholtz-hzi.de:10033/6211362019-08-30T11:27:16Zcom_10033_311624com_10033_6839com_10033_620618col_10033_311625col_10033_620619
00925njm 22002777a 4500
dc
Rox, Katharina
author
Jansen, Rolf
author
Loof, Torsten G.
author
Gillen, Christine M.
author
Bernecker, Steffen
author
Walker, Mark J.
author
Chhatwal, Gursharan Singh
author
Müller, Rolf
author
2017-09-18
In contrast to mild infections of Group A Streptococcus (GAS) invasive infections of GAS still pose a serious health hazard: GAS disseminates from sterile sites into the blood stream or deep tissues and causes sepsis or necrotizing fasciitis. In this case antibiotics do not provide an effective cure as the bacteria are capable to hide from them very quickly. Therefore, new remedies are urgently needed. Starting from a myxobacterial natural products screening campaign, we identified two fatty acids isolated from myxobacteria, linoleic and palmitoleic acid, specifically blocking streptokinase-mediated activation of plasminogen and thereby preventing streptococci from hijacking the host’s plasminogen/plasmin system. This activity is not inherited by other fatty acids such as oleic acid and is not attributable to the killing of streptococci. Moreover, both fatty acids are superior in their inhibitory properties compared to two clinically used drugs (tranexamic or ε-amino caproic acid) as they show 500–1000 fold lower IC50 values. Using a humanized plasminogen mouse model mimicking the clinical situation of a local GAS infection that becomes systemic, we demonstrate that these fatty acids ameliorate invasive GAS infection significantly. Consequently, linoleic and palmitoleic acid are possible new options to combat GAS invasive diseases.
Linoleic and palmitoleic acid block streptokinase-mediated plasminogen activation and reduce severity of invasive group A streptococcal infection 2017, 7 (1) Scientific Reports
2045-2322
10.1038/s41598-017-11276-z
http://hdl.handle.net/10033/621136
Scientific Reports
Linoleic and palmitoleic acid block streptokinase-mediated plasminogen activation and reduce severity of invasive group A streptococcal infection
oai:repository.helmholtz-hzi.de:10033/6211622019-08-30T11:28:51Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Debarry, Jennifer
author
Cornberg, Markus
author
Manns, Michael P
author
2017-01
Despite an available vaccine and efficient treatment for hepatitis B virus (HBV) infection, chronic HBV infection still remains a major global threat, and one of the top 20 causes of human mortality worldwide. One of the major challenges in controlling HBV infection is the high number of undiagnosed chronic carriers and the lack of access to prophylaxis and treatment in several parts of the world. We discuss relevant barriers that need to be overcome to achieve global control of HBV infection and make eradication possible. Most important, vaccination must be scaled-up to lower the risk of vertical transmission and decrease the number of new infections, and comprehensive screening programs must be linked to care to obtain a better rate of diagnosis and treatment. This can probably only be achieved if sustainable funding is available. We therefore emphasize the importance of making the management of viral hepatitis a global health priority.
Challenges in warranting access to prophylaxis and therapy for hepatitis B virus infection. 2017, 37 Suppl 1:67-72 Liver Int.
1478-3231
28052625
10.1111/liv.13320
http://hdl.handle.net/10033/621162
Liver international : official journal of the International Association for the Study of the Liver
Challenges in warranting access to prophylaxis and therapy for hepatitis B virus infection.
oai:repository.helmholtz-hzi.de:10033/6211682019-08-30T11:34:43Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Manns, Michael P
author
2017-10
Hepatitis D virus in Africa: several unmet needs. 2017, 5 (10):e953-e954 Lancet Glob Health
2214-109X
28911754
10.1016/S2214-109X(17)30345-5
http://hdl.handle.net/10033/621168
The Lancet. Global health
Hepatitis D virus in Africa: several unmet needs.
oai:repository.helmholtz-hzi.de:10033/6212192019-08-30T11:35:36Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Nishanth, G
author
Wolleschak, D
author
Fahldieck, C
author
Fischer, T
author
Mullally, A
author
Perner, F
author
Schnöder, T M
author
Just, S
author
Heidel, F H
author
Schlüter, D
author
2017-04
Gain of function in Jak2V617F-positive T-cells. 2017, 31 (4):1000-1003 Leukemia
1476-5551
28074070
10.1038/leu.2017.6
http://hdl.handle.net/10033/621219
Leukemia
Gain of function in Jak2V617F-positive T-cells.
oai:repository.helmholtz-hzi.de:10033/6212372019-08-30T11:35:10Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Manns, Michael P
author
2016-12
Future Organization of Clinical Research in Germany: The Road to the "German Centre for Digestive Health" (GCDH). 2016, 54 (12):1293-1295 Z Gastroenterol
1439-7803
27936478
10.1055/s-0042-118288
http://hdl.handle.net/10033/621237
Zeitschrift fur Gastroenterologie
Future Organization of Clinical Research in Germany: The Road to the "German Centre for Digestive Health" (GCDH).
oai:repository.helmholtz-hzi.de:10033/6212382019-08-30T11:35:10Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Manns, Michael P
author
Jaeckel, Elmar
author
Taubert, Richard
author
2017-11-08
Budesonide in Autoimmune Hepatitis: The Right Drug at the Right Time for the Right Patient. 2017 Clin. Gastroenterol. Hepatol.
1542-7714
29128475
10.1016/j.cgh.2017.11.003
http://hdl.handle.net/10033/621238
Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association
Budesonide in Autoimmune Hepatitis: The Right Drug at the Right Time for the Right Patient.
oai:repository.helmholtz-hzi.de:10033/6213602019-08-30T11:33:57Zcom_10033_620533com_10033_311624com_10033_6839com_10033_338554col_10033_621787col_10033_620534col_10033_311625
00925njm 22002777a 4500
dc
Rahim, Muhammad Imran
author
Weizbauer, Andreas
author
Evertz, Florian
author
Hoffmann, Andrea
author
Rohde, M
author
Glasmacher, Birgit
author
Windhagen, Henning
author
Gross, Gerhard
author
Seitz, Jan-Marten
author
Mueller, Peter P
author
2017
Magnesium alloys are presently under investigation as promising biodegradable implant materials with osteoconductive properties. To study the molecular mechanisms involved, the potential contribution of soluble magnesium corrosion products to the stimulation of osteoblastic cell differentiation was examined. However, no evidence for the stimulation of osteoblast differentiation could be obtained when cultured mesenchymal precursor cells were differentiated in the presence of metallic magnesium or in cell culture medium containing elevated magnesium ion levels. Similarly, in soft tissue no bone induction by metallic magnesium or by the corrosion product magnesium hydroxide could be observed in a mouse model. Motivated by the comparatively rapid accumulation solid corrosion products physicochemical processes were examined as an alternative mechanism to explain the stimulation of bone growth by magnesium-based implants. During exposure to physiological solutions a structured corrosion coat formed on magnesium whereby the elements calcium and phosphate were enriched in the outermost layer which could play a role in the established biocompatible behavior of magnesium implants. When magnesium pins were inserted into avital bones, corrosion lead to increases in the pull out force, suggesting that the expanding corrosion layer was interlocking with the surrounding bone. Since mechanical stress is a well-established inducer of bone growth, volume increases caused by the rapid accumulation of corrosion products and the resulting force development could be a key mechanism and provide an explanation for the observed stimulatory effects of magnesium-based implants in hard tissue. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 697-709, 2017.
Differential magnesium implant corrosion coat formation and contribution to bone bonding. 2017, 105 (3):697-709 J Biomed Mater Res A
1552-4965
27770566
10.1002/jbm.a.35943
http://hdl.handle.net/10033/621360
Journal of biomedical materials research. Part A
Differential magnesium implant corrosion coat formation and contribution to bone bonding.
oai:repository.helmholtz-hzi.de:10033/6214092019-08-30T11:32:37Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Strunz, Benedikt
author
Hengst, Julia
author
Deterding, Katja
author
Manns, Michael P
author
Cornberg, Markus
author
Ljunggren, Hans-Gustaf
author
Wedemeyer, Heiner
author
Björkström, Niklas K
author
2018-06-11
Diversity is a central requirement for the immune system's capacity to adequately clear a variety of different infections. As such, natural killer (NK) cells represent a highly diverse population of innate lymphocytes important in the early response against viruses. Yet, the extent to which a chronic pathogen affects NK cell diversity is largely unknown. Here we study NK cell functional diversification in chronic hepatitis C virus (HCV) infection. High-dimensional flow cytometer assays combined with stochastic neighbor embedding analysis reveal that chronic HCV infection induces functional imprinting on human NK cells that is largely irreversible and persists long after successful interventional clearance of the virus. Furthermore, HCV infection increases inter-individual, but decreases intra-individual, NK cell diversity. Taken together, our results provide insights into how the history of infections affects human NK cell diversity.
2041-1723
29891939
10.1038/s41467-018-04685-9
http://hdl.handle.net/10033/621409
Chronic hepatitis C virus infection irreversibly impacts human natural killer cell repertoire diversity.
oai:repository.helmholtz-hzi.de:10033/6214222019-08-30T11:33:27Zcom_10033_311624com_10033_6839com_10033_620636col_10033_311625col_10033_620638
00925njm 22002777a 4500
dc
Kremling, Andreas
author
Goehler, Anna
author
Jahreis, Knut
author
Nees, Markus
author
Auerbach, Benedikt
author
Schmidt-Heck, Wolfgang
author
Kökpinar, Oznur
author
Geffers, Robert
author
Rinas, Ursula
author
Bettenbrock, Katja
author
2012-11-12
Metabolism and signalling are tightly coupled in bacteria. Combining several theoretical approaches, a core model is presented that describes transcriptional and allosteric control of glycolysis in Escherichia coli. Experimental data based on microarrays, signalling components and extracellular metabolites are used to estimate kinetic parameters. A newly designed strain was used that adjusts the incoming glucose flux into the system and allows a kinetic analysis. Based on the results, prediction for intracelluar metabolite concentrations over a broad range of the growth rate could be performed and compared with data from literature.
2218-1989
24957765
10.3390/metabo2040844
http://hdl.handle.net/10033/621422
Analysis and Design of Stimulus Response Curves of E. coli.
oai:repository.helmholtz-hzi.de:10033/6214272018-07-19T17:28:27Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Strunz, Benedikt
author
Hengst, Julia
author
Deterding, Katja
author
Manns, Michael P
author
Cornberg, Markus
author
Ljunggren, Hans-Gustaf
author
Wedemeyer, Heiner
author
Björkström, Niklas K
author
2018-06-11
Diversity is a central requirement for the immune system's capacity to adequately clear a variety of different infections. As such, natural killer (NK) cells represent a highly diverse population of innate lymphocytes important in the early response against viruses. Yet, the extent to which a chronic pathogen affects NK cell diversity is largely unknown. Here we study NK cell functional diversification in chronic hepatitis C virus (HCV) infection. High-dimensional flow cytometer assays combined with stochastic neighbor embedding analysis reveal that chronic HCV infection induces functional imprinting on human NK cells that is largely irreversible and persists long after successful interventional clearance of the virus. Furthermore, HCV infection increases inter-individual, but decreases intra-individual, NK cell diversity. Taken together, our results provide insights into how the history of infections affects human NK cell diversity.
2041-1723
29891939
10.1038/s41467-018-04685-9
http://hdl.handle.net/10033/621427
Chronic hepatitis C virus infection irreversibly impacts human natural killer cell repertoire diversity.
oai:repository.helmholtz-hzi.de:10033/6214392019-08-30T11:34:45Zcom_10033_311624com_10033_6839com_10033_620601com_10033_620652col_10033_620672col_10033_311625col_10033_620602
00925njm 22002777a 4500
dc
Selvakumar, Tharini A
author
Bhushal, Sudeep
author
Kalinke, Ulrich
author
Wirth, Dagmar
author
Hauser, Hansjörg
author
Köster, Mario
author
Hornef, Mathias W
author
2017-01-01
Type I (α and β) and type III (λ) interferons (IFNs) induce the expression of a large set of antiviral effector molecules
1664-3224
29085367
10.3389/fimmu.2017.01302
http://hdl.handle.net/10033/621439
gastrointestinal tract
interferon-lambda
interleukin 28 receptor
intestinal epithelium
transcription
Identification of a Predominantly Interferon-λ-Induced Transcriptional Profile in Murine Intestinal Epithelial Cells.
oai:repository.helmholtz-hzi.de:10033/6214402019-08-30T11:35:10Zcom_10033_311624com_10033_6839com_10033_311308col_10033_311625col_10033_620721
00925njm 22002777a 4500
dc
Quaas, Bastian
author
Burmeister, Laura
author
Li, Zhaopeng
author
NIMTZ, MANFRED
author
Hoffmann, Andrea
author
Rinas, Ursula
author
13595113
10.1016/j.procbio.2018.02.001
http://hdl.handle.net/10033/621440
Properties of dimeric, disulfide-linked rhBMP-2 recovered from E. coli derived inclusion bodies by mild extraction or chaotropic solubilization and subsequent refolding
oai:repository.helmholtz-hzi.de:10033/6214872019-08-30T11:27:45Zcom_10033_311624com_10033_6839com_10033_311308col_10033_311625col_10033_620721
00925njm 22002777a 4500
dc
Roth, Gustavo
author
Vanz, Ana Letícia
author
Lünsdorf, Heinrich
author
Nimtz, Manfred
author
Rinas, Ursula
author
2018-08-09
Secretory recombinant protein production with Pichia (syn. Komagataella) pastoris is commonly associated with the induction of an unfolded protein response (UPR) usually apparent through increased intracellular levels of endoplasmic reticulum (ER) resident chaperones such as Kar2/Bip. During methanol-induced secretory production of an insulin precursor (IP) under industrially relevant fed-batch conditions the initially high level of intracellular Kar2/Bip after batch growth on glycerol unexpectedly declined in the following methanol fed-batch phase misleadingly suggesting that IP production had a low impact on UPR activation. Analysis of the protein production independent level of Kar2/Bip revealed that high Kar2/Bip levels were reached in the exponential growth phase of glycerol batch cultures followed by a strong decline of Kar2/Bip during entry into stationary phase. Ultra-structural cell morphology studies revealed autophagic processes (e.g. ER phagy) at the end of the glycerol batch phase most likely responsible for the degradation of ER resident chaperones such as Kar2/Bip. The pre-induction level of Kar2/Bip did not affect the IP secretion efficiency in the subsequent methanol-induced IP production phase. During growth on methanol intracellular Kar2/Bip levels declined in IP producing and non-producing host cells. However, extracellular accumulation of Kar2/Bip was observed in IP-producing cultures but not in non-producing controls. Most importantly, the majority of the extracellular Kar2/Bip accumulated in the culture supernatant of IP producing cells as truncated protein (approx. 35 kDa). Rapid growth leads to higher basal levels of the major UPR marker protein Kar2/Bip independent of recombinant protein production. Entry into stationary phase or slower growth on poorer substrate, e.g. methanol, leads to a lower basal Kar2/Bip level. Methanol-induced secretory IP production elicits a strong UPR activation which counteracts the reduced UPR during slow growth on methanol. The major ER chaperone Kar2/Bip is found together with recombinant IP in the culture medium where full-length Kar2/Bip accumulates in addition to large amounts of truncated Kar2/Bip. Thus, for judging UPR activating properties of the produced protein it is important to additionally analyze the medium not only for intact Kar2/Bip but also for truncated versions of this UPR reporter protein.
1475-2859
30092809
10.1186/s12934-018-0970-3
http://hdl.handle.net/10033/621487
Autophagy
Kar2/Bip
Pichia pastoris
Unfolded protein response
Fate of the UPR marker protein Kar2/Bip and autophagic processes in fed-batch cultures of secretory insulin precursor producing Pichia pastoris.
oai:repository.helmholtz-hzi.de:10033/6215192019-08-30T11:29:45Zcom_10033_311624com_10033_6839com_10033_620636col_10033_311625col_10033_620637
00925njm 22002777a 4500
dc
Müsken, Mathias
author
Pawar, Vinay
author
Schwebs, Timo
author
Bähre, Heike
author
Felgner, Sebastian
author
Weiss, Siegfried
author
Häussler, Susanne
author
2018-10-08
Biofilm-residing bacteria embedded in an extracellular matrix are protected from diverse
physico-chemical insults. In addition to the general recalcitrance of biofilm-bacteria, high
bacterial loads in biofilm-associated infections significantly diminishes the efficacy of
antimicrobials due to a low per-cell antibiotic concentration. Accordingly, present antimicrobial
treatment protocols, that have been established to serve the eradication of acute infections, fail
to clear biofilm-associated chronic infections. In the present study, we applied automated
confocal microscopy on Pseudomonas aeruginosa to monitor dynamic killing of biofilm-grown
bacteria by tobramycin and colistin in real-time. We revealed that the time required for
surviving bacteria to repopulate the biofilm could be taken as measure for effectiveness of the
antimicrobial treatment. It depends on the: i) nature and concentration of the antibiotic, ii)
duration of antibiotic treatment; iii) application as mono or combination therapy and iv) time
intervals of drug administration. The vicious cycle of killing and repopulation of biofilm
bacteria could also be broken in an in vivo model system by applying successive antibiotic
dosages with time intervals that do not allow full reconstitution of the biofilm communities.
Treatment regimens that consider the important aspects of antimicrobial killing kinetics bear
the potential to improve control of biofilm regrowth. This is an important and underestimated
factor that is bound to ensure sustainable treatment success of chronic infections.
1098-6596
30297365
10.1128/AAC.01635-18
http://hdl.handle.net/10033/621519
Breaking the vicious cycle of antibiotic killing and regrowth of biofilm-residing .
oai:repository.helmholtz-hzi.de:10033/6215452019-08-30T11:30:26Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Meiser, Johannes
author
Kraemer, Lisa
author
Jaeger, Christian
author
Madry, Henning
author
Link, Andreas
author
Lepper, Philipp M
author
Hiller, Karsten
author
Schneider, Jochen G
author
2018-08-14
Itaconic acid is produced by mammalian leukocytes upon pro-inflammatory activation. It appears to inhibit bacterial growth and to rewire the metabolism of the host cell by inhibiting succinate dehydrogenase. Yet, it is unknown whether itaconic acid acts only intracellularly, locally in a paracrine fashion, or whether it is even secreted from the inflammatory cells at meaningful levels in peripheral blood of patients with severe inflammation or sepsis. The aim of this study was to determine the release rate of itaconic acid from pro-inflammatory activated macrophages
1949-2553
30181801
10.18632/oncotarget.25956
http://hdl.handle.net/10033/621545
Immunology
biomarker
inflammation
itaconic acid
metabolism
sepsis
Itaconic acid indicates cellular but not systemic immune system activation.
oai:repository.helmholtz-hzi.de:10033/6215672019-08-30T11:29:43Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Broglia, Laura
author
Materne, Solange
author
Lécrivain, Anne-Laure
author
Hahnke, Karin
author
Le Rhun, Anaïs
author
Charpentier, Emmanuelle
author
2018-01-01
Endoribonuclease Y (RNase Y) is a crucial regulator of virulence in Gram-positive bacteria. In the human pathogen Streptococcus pyogenes, RNase Y is required for the expression of the major secreted virulence factor streptococcal pyrogenic exotoxin B (SpeB), but the mechanism involved in this regulation remains elusive. Here, we demonstrate that the 5' untranslated region of speB mRNA is processed by several RNases including RNase Y. In particular, we identify two RNase Y cleavage sites located downstream of a guanosine (G) residue. To assess whether this nucleotide is required for RNase Y activity in vivo, we mutated it and demonstrate that the presence of this G residue is essential for the processing of the speB mRNA 5' UTR by RNase Y. Although RNase Y directly targets and processes speB, we show that RNase Y-mediated regulation of speB expression occurs primarily at the transcriptional level and independently of the processing in the speB mRNA 5' UTR. To conclude, we demonstrate for the first time that RNase Y processing of an mRNA target requires the presence of a G. We also provide new insights on the speB 5' UTR and on the role of RNase Y in speB regulation.
1555-8584
30290721
10.1080/15476286.2018.1532253
http://hdl.handle.net/10033/621567
5′ untranslated region
RNase Y
Streptococcus pyogenes
speB
transcriptional regulation
virulence
RNase Y-mediated regulation of the streptococcal pyrogenic exotoxin B.
oai:repository.helmholtz-hzi.de:10033/6215702019-08-30T11:29:43Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Jupa, K.L.
author
Manns, M.P.
author
Jäckel, E
author
Epidemiologie
Die nicht-alkoholische Fettlebererkrankung
(»non-alcoholic fatty liver disease«, NAFLD) ist
mittlerweile eine der häufigsten Lebererkrankung weltweit mit einer mittleren Prävalenz von
20% weltweit und bis zu 30% in Europa [1]. Als
Hauptrisikofaktoren gelten einerseits verschiedene Umweltfaktoren, insbesondere Bewegungsmangel und falsche Ernährung sowie andererseits
die verschiedenen Aspekte des metabolischen
Syndroms: Adipositas, Fettstoffwechselstörungen sowie Insulinresistenz und Diabetes mellitus Typ 2. Aus diesem Grund wird die Fettleber
mittlerweile auch als hepatische Komponente
des metabolischen Syndroms bezeichnet. Neben den Umweltfaktoren konnte auch gezeigt
werden, dass eine genetische Prädisposition im
Sinne von einem Adiponutrin Polymorphismus
(»patatin-like phospholipase domain containing
3«, PNPLA3 [2, 3] zu einem gehäuften Auftreten
einer Fettlebererkrankung führen kann.
0494464X
http://hdl.handle.net/10033/621570
Tägliche Praxis
Die nicht-alkoholische Fettlebererkrankung : Die Rolle der Leber im metabolischen Syndrom
oai:repository.helmholtz-hzi.de:10033/6215842019-08-30T11:30:27Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Taubert, R.
author
Manns, M. P.
author
Gegliederte Zusammenfassung
Hintergrund: Die Autoimmunhepatitis (AIH) ist zwar eine seltene Erkrankung zeigt
aber wie andere Autoimmunerkrankungen in der westlichen Welt eine ansteigende
Inzidenz und hat unbehandelt einen schlechten natürlichen Verlauf.
Ziel der Arbeit (Fragestellung): Darstellung des aktuellen Kenntnisstands zur
Pathogenese, Diagnostik und Behandlung der AIH.
Material und Methoden: Zusammenfassung der gültigen nationalen sowie
internationalen Leitlinien und exemplarischer aktuell publizierter Studien.
Ergebnisse und Diskussion: Die Therapie der AIH aus Prednisolon +/- Azathioprin
war beginnend in den 1960 Jahren die erste medikamentöse Therapie einer
Lebererkrankung, die die Lebenserwartung nachweislich verbessern konnte. Seit
2011 Jahren ist zusätzlich Budesonid für AIH-Patienten ohne Leberzirrhose als
alternatives Steroid mit weniger systemischen Nebenwirkungen zugelassen.
Abgesehen davon hat sich die initiale Erstlinientherapie der AIH in den letzten 40
Jahren nicht grundlegend verändert. Das Therapieziel der kompletten biochemischen
Remission wird bei ca. 70-80% der Patienten erreicht. Bei hohen Rückfallraten trotz
lang anhaltender biochemischer und histologischer Remission ist bei den meisten
Patienten eine lebenslange Therapie notwendig. Bisherige Zweitlinientherapien
beruhen vor allem auf retrospektiven Studienergebnissen und daher fehlen
einheitliche Empfehlungen zur Zweitlinientherapie von den internationalen
Fachgesellschaften. Ebenso ist keine Zweitlinientherapie von den
Zulassungsbehörden wie FDA oder EMA zugelassen.
1861-9681
1861-969X
10.1007/s11377-018-0251-z
http://hdl.handle.net/10033/621584
Autoimmune hepatitis: From the initial therapy to the differentiated approach [Autoimmunhepatitis: Von der ersten Therapie zum differenzierten Vorgehen]
oai:repository.helmholtz-hzi.de:10033/6216122019-08-30T11:30:28Zcom_10033_311624com_10033_6839com_10033_311308col_10033_311625col_10033_620721
00925njm 22002777a 4500
dc
Brennecke, Anne-Margarete
author
Düber, Sandra
author
Roy, Bishnudeo
author
Thomsen, Irene
author
Garbe, Annette I
author
Klawonn, Frank
author
Pabst, Oliver
author
Kretschmer, Karsten
author
Weiss, Siegfried
author
2018-01-01
We employed the B-Indu-Rag1 model in which the coding exon of recombination-activating gene 1 (Rag1) is inactivated by inversion. It is flanked by inverted loxP sites. Accordingly, B cell development is stopped at the pro/pre B-I cell precursor stage. A B cell-specific Cre recombinase fused to a mutated estrogen receptor allows the induction of RAG1 function and B cell development by application of Tamoxifen. Since Rag1 function is recovered in a non-self-renewing precursor cell, only single waves of development can be induced. Using this system, we could determine that B cells minimally require 5 days to undergo development from pro/preB-I cells to the large and 6 days to the small preB-II cell stage. First immature transitional (T) 1 and T2 B cells could be detected in the bone marrow at day 6 and day 7, respectively, while their appearance in the spleen took one additional day. We also tested a contribution of adult bone marrow to the pool of B-1 cells. Sublethally irradiated syngeneic WT mice were adoptively transferred with bone marrow of B-Indu-Rag1 mice and B cell development was induced after 6 weeks. A significant portion of donor derived B-1 cells could be detected in such adult mice. Finally, early VH gene usage was tested after induction of B cell development. During the earliest time points the VH genes proximal to D/J were found to be predominantly rearranged. At later time points, the large family of the most distal VH prevailed.
1664-3224
30429851
10.3389/fimmu.2018.02483
http://hdl.handle.net/10033/621612
B cell development
B-2/B-1a/B-1b
CSR
RAG
T-dependent/-independent
VH usage
antibodies
bone marrow
Induced B Cell Development in Adult Mice.
oai:repository.helmholtz-hzi.de:10033/6216152020-08-12T13:54:07Zcom_10033_311624com_10033_6839com_10033_620626com_10033_620636col_10033_311625col_10033_620629col_10033_620637
00925njm 22002777a 4500
dc
Lorenz, Anne
author
Preuße, Matthias
author
Bruchmann, Sebastian
author
Pawar, Vinay
author
Grahl, Nora
author
Pils, Marina C
author
Nolan, Laura M
author
Filloux, Alain
author
Weiss, Siegfried
author
Häussler, Susanne
author
2018-11-08
Pseudomonas aeruginosa is an environmental microorganism and a causative agent of diverse acute and chronic, biofilm-associated infections. Advancing research-based knowledge on its adaptation to conditions within the human host is bound to reveal novel strategies and targets for therapeutic intervention. Here, we investigated the traits that P. aeruginosa PA14 as well as a virulence attenuated ΔlasR mutant need to survive in selected murine infection models. Experimentally, the genetic programs that the bacteria use to adapt to biofilm-associated versus acute infections were dissected by passaging transposon mutant libraries through mouse lungs (acute) or mouse tumours (biofilm-infection). Adaptive metabolic changes of P. aeruginosa were generally required during both infection processes. Counter-selection against flagella expression was observed during acute lung infections. Obviously, avoidance of flagella-mediated activation of host immunity is advantageous for the wildtype bacteria. For the ΔlasR mutant, loss of flagella did not confer a selective advantage. Apparently, other pathogenesis mechanisms are active in this virulence attenuated strain. In contrast, the infective process of P. aeruginosa in the chronic biofilm model apparently required expression of flagellin. Together, our findings imply that the host immune reactions against the infectious agent are very decisive for acuteness and duration of the infectious disease. They direct disease outcome.
1462-2920
30411474
10.1111/1462-2920.14468
http://hdl.handle.net/10033/621615
Importance of flagella in acute and chronic Pseudomonas aeruginosa infections.
oai:repository.helmholtz-hzi.de:10033/6216172019-08-30T11:30:28Zcom_10033_311624com_10033_6839com_10033_620589col_10033_311625col_10033_620596
00925njm 22002777a 4500
dc
Khera, Tanvi
author
Behrendt, Patrick
author
Bankwitz, Dorothea
author
Brown, Richard J P
author
Todt, Daniel
author
Doepke, Mandy
author
Ghafoor Khan, Abdul
author
Schulze, Kai
author
Law, John
author
Logan, Michael
author
Hockman, Darren
author
Wong, Jason Alexander Ji-Xhin
author
Dold, Leona
author
Gonzalez-Motos, Victor
author
Spengler, Ulrich
author
Viejo-Borbolla, Abel
author
Ströh, Luisa
author
Krey, Thomas
author
Tarr, Alexander W
author
Steinmann, Eike
author
Manns, Michael P
author
Klein, Florian
author
Guzman, Carlos A
author
Marcotrigiano, Joseph
author
Houghton, Michael
author
Pietschmann, Thomas
author
2018-11-12
Induction of cross-reactive antibodies targeting conserved epitopes of the envelope proteins E1E2 is a key requirement for an HCV vaccine. Conserved epitopes like the viral CD81-binding site are targeted by rare broadly neutralizing antibodies. However, these viral segments are occluded by variable regions and glycans. We aimed to identify antigens exposing conserved epitopes and to characterize their immunogenicity. We created HCV variants with mutated glycosylation sites and/or hypervariable region 1 (HVR1). Exposure of the CD81 binding site and conserved epitopes was quantified by soluble CD81 and antibody interaction and neutralization assays. E2 or E1-E2 heterodimers with mutations causing epitope exposure were used to immunize mice. Vaccine-induced antibodies were examined and compared with patient-derived antibodies. Mutant viruses bound soluble CD81 and antibodies targeting the CD81 binding site with enhanced efficacy. Mice immunized with E2 or E1E2 heterodimers incorporating these modifications mounted strong, cross-binding, and non-interfering antibodies. E2-induced antibodies neutralized the autologous virus but they were not cross-neutralizing. Viruses lacking the HVR1 and selected glycosylation sites expose the CD81 binding site and cross-neutralization antibody epitopes. Recombinant E2 proteins carrying these modifications induce strong cross-binding but not cross-neutralizing antibodies.
1600-0641
30439392
10.1016/j.jhep.2018.11.003
http://hdl.handle.net/10033/621617
Antibodies
Glycoproteins
HCV
Immunogen
Recombinant proteins
Functional and immunogenic characterization of diverse HCV glycoprotein E2 variants.
oai:repository.helmholtz-hzi.de:10033/6216212019-08-30T11:33:02Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Vogel, Katrin
author
Pierau, Mandy
author
Arra, Aditya
author
Lampe, Karen
author
Schlueter, Dirk
author
Arens, Christoph
author
Brunner-Weinzierl, Monika C
author
2018-11-15
The origin of human T-cell responses against fungal pathogens early in life is not clearly understood. Here, we show that antifungal T-cell responses are vigorously initiated within the first years of life against lysates and peptides of Candida albicans or Aspergillus fumigatus, presented by autologous monocytes. The neonatal responding T-cell pool consists of 20 different TCR-V
2045-2322
30442915
10.1038/s41598-018-35161-5
http://hdl.handle.net/10033/621621
Developmental induction of human T-cell responses against Candida albicans and Aspergillus fumigatus.
oai:repository.helmholtz-hzi.de:10033/6216292019-08-30T11:28:47Zcom_10033_311624com_10033_6839com_10033_620652com_10033_338554col_10033_621787col_10033_620672col_10033_311625
00925njm 22002777a 4500
dc
Ullah, Sami
author
Seidel, Katja
author
Türkkan, Sibel
author
Warwas, Dawid Peter
author
Dubich, Tatyana
author
Rohde, Manfred
author
Hauser, Hansjörg
author
Behrens, Peter
author
Kirschning, Andreas
author
Köster, Mario
author
Wirth, Dagmar
author
2018-12-23
Targeted delivery of drugs is a major challenge in treatment of diverse diseases. Systemically administered drugs demand high doses and are accompanied by poor selectivity and side effects on non-target cells. Here, we introduce a new principle for targeted drug delivery. It is based on macrophages as transporters for nanoparticle-coupled drugs as well as controlled release of drugs by hyperthermia mediated disruption of the cargo cells and simultaneous deliberation of nanoparticle-linked drugs. Hyperthermia is induced by an alternating electromagnetic field (AMF) that induces heat from silica-coated superparamagnetic iron oxide nanoparticles (SPIONs). We show proof-of-principle of controlled release by the simultaneous disruption of the cargo cells and the controlled, AMF induced release of a toxin, which was covalently linked to silica-coated SPIONs via a thermo-sensitive linker. Cells that had not been loaded with SPIONs remain unaffected. Moreover, in a 3D co-culture model we demonstrate specific killing of associated tumour cells when employing a ratio as low as 1:40 (SPION-loaded macrophage: tumour cells). Overall, our results demonstrate that AMF induced drug release from macrophage-entrapped nanoparticles is tightly controlled and may be an attractive novel strategy for targeted drug release.
1873-4995
30586597
10.1016/j.jconrel.2018.12.040
http://hdl.handle.net/10033/621629
Cell based drug delivery
Controlled drug delivery, 3D tumour model
Hyperthermia
Macrophages
Magnetic silica nanoparticles
Macrophage entrapped silica coated superparamagnetic iron oxide particles for controlled drug release in a 3D cancer model.
oai:repository.helmholtz-hzi.de:10033/6216712019-08-30T11:30:30Zcom_10033_311624com_10033_6839com_10033_620857com_10033_620652com_10033_620618col_10033_620672col_10033_311625col_10033_620858col_10033_620619
00925njm 22002777a 4500
dc
Dubich, Tatyana
author
Lieske, Anna
author
Santag, Susann
author
Beauclair, Guillaume
author
Rückert, Jessica
author
Herrmann, Jennifer
author
Gorges, Jan
author
Büsche, Guntram
author
Kazmaier, Uli
author
Hauser, Hansjörg
author
Stadler, Marc
author
Schulz, Thomas F
author
Wirth, Dagmar
author
2019-01-04
Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent of Kaposi's sarcoma (KS), a tumor of endothelial origin predominantly affecting immunosuppressed individuals. Up to date, vaccines and targeted therapies are not available. Screening and identification of anti-viral compounds are compromised by the lack of scalable cell culture systems reflecting properties of virus-transformed cells in patients. Further, the strict specificity of the virus for humans limits the development of in vivo models. In this study, we exploited a conditionally immortalized human endothelial cell line for establishment of in vitro 2D and 3D KSHV latency models and the generation of KS-like xenograft tumors in mice. Importantly, the invasive properties and tumor formation could be completely reverted by purging KSHV from the cells, confirming that tumor formation is dependent on the continued presence of KSHV, rather than being a consequence of irreversible transformation of the infected cells. Upon testing a library of 260 natural metabolites, we selected the compounds that induced viral loss or reduced the invasiveness of infected cells in 2D and 3D endothelial cell culture systems. The efficacy of selected compounds against KSHV-induced tumor formation was verified in the xenograft model. Together, this study shows that the combined use of anti-viral and anti-tumor assays based on the same cell line is predictive for tumor reduction in vivo and therefore allows faithful selection of novel drug candidates against Kaposi's sarcoma. KEY MESSAGES: Novel 2D, 3D, and xenograft mouse models mimic the consequences of KSHV infection. KSHV-induced tumorigenesis can be reverted upon purging the cells from the virus. A 3D invasiveness assay is predictive for tumor reduction in vivo. Chondramid B, epothilone B, and pretubulysin D diminish KS-like lesions in vivo.
1432-1440
30610257
10.1007/s00109-018-01733-1
http://hdl.handle.net/10033/621671
3D culture system
Drug validation
Humanized mouse model
KSHV
Novel anti-viral drugs
An endothelial cell line infected by Kaposi's sarcoma-associated herpes virus (KSHV) allows the investigation of Kaposi's sarcoma and the validation of novel viral inhibitors in vitro and in vivo.
oai:repository.helmholtz-hzi.de:10033/6217062019-08-30T11:32:40Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Zahid, Maria
author
Rinas, Ursula
author
2019-01-01
Virus-like particle (VLP)-based vaccines have been in the market since decades
for preventing viral infection and have proven their usefulness also in other
areas of biotechnology. Here, we describe in detail simple small-scale production
and purification procedures for the generation of hepatitis B surface antigen
(HBsAg) VLPs using Pichia pastoris as expression host. This protocol may also be
applicable with variations to other HBsAg-based VLPs additionally carrying
antigens of other pathogens.
Methods Mol Biol. 2019;1923:309-322. doi: 10.1007/978-1-4939-9024-5_14.
1940-6029
30737747
10.1007/978-1-4939-9024-5_14
http://hdl.handle.net/10033/621706
Methods in molecular biology
HBsAg
Pichia pastoris
Vaccine
Virus
Virus-like particle
Guidelines for Small-Scale Production and Purification of Hepatitis B Surface Antigen Virus-Like Particles from Recombinant Pichia pastoris.
oai:repository.helmholtz-hzi.de:10033/6217082019-08-30T11:35:13Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Tsay, Hsin-Chieh
author
Yuan, Qinggong
author
Balakrishnan, Asha
author
Kaiser, Marina
author
Möbus, Selina
author
Kozdrowska, Emilia
author
Farid, Marwa
author
Tegtmeyer, Pia-Katharina
author
Borst, Katharina
author
Vondran, Florian W R
author
Kalinke, Ulrich
author
Kispert, Andreas
author
Manns, Michael P
author
Ott, Michael
author
Sharma, Amar Deep
author
2018-12-22
Fibrosis, a cardinal feature of a dysfunctional liver, significantly contributes to the ever-increasing mortality due to end-stage chronic liver diseases. The crosstalk between hepatocytes and hepatic stellate cells (HSCs) plays a key role in the progression of fibrosis. Although ample efforts have been devoted to elucidate the functions of HSCs during liver fibrosis, the regulatory functions of hepatocytes remain elusive. Using an unbiased functional microRNA (miRNA) screening, we investigated the ability of hepatocytes to regulate fibrosis by fine-tuning gene expression via miRNA modulation. The in vivo functional analyses were performed by inhibiting miRNA in hepatocytes using adeno-associated virus in carbon-tetrachloride- and 3,5-di-diethoxycarbonyl-1,4-dihydrocollidine-induced liver fibrosis. Blocking miRNA-221-3p function in hepatocytes during chronic liver injury facilitated recovery of the liver and faster resolution of the deposited extracellular matrix. Furthermore, we demonstrate that reduced secretion of C-C motif chemokine ligand 2, as a result of post-transcriptional regulation of GNAI2 (G protein alpha inhibiting activity polypeptide 2) by miRNA-221-3p, mitigates liver fibrosis. Collectively, miRNA modulation in hepatocytes, an easy-to-target cell type in the liver, may serve as a potential therapeutic approach for liver fibrosis.
1600-0641
30582979
10.1016/j.jhep.2018.12.016
http://hdl.handle.net/10033/621708
Journal of Hepatology
Chemokine ligands
Fibrosis
Hepatic stellate cells
miRNA
miRNA-221-3p
Hepatocyte-specific suppression of microRNA-221-3p mitigates liver fibrosis.
oai:repository.helmholtz-hzi.de:10033/6217272019-08-30T11:28:51Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Groenewold, Maike K
author
Hebecker, Stefanie
author
Fritz, Christiane
author
Czolkoss, Simon
author
Wiesselmann, Milan
author
Heinz, Dirk W
author
Jahn, Dieter
author
Narberhaus, Franz
author
Aktas, Meriyem
author
Moser, Jürgen
author
2019-01-01
Agrobacterium tumefaciens transfers oncogenic T-DNA via the type IV secretion system (T4SS) into plants causing tumor formation. The acvB gene encodes a virulence factor of unknown function required for plant transformation. Here we specify AcvB as a periplasmic lysyl-phosphatidylglycerol (L-PG) hydrolase, which modulates L-PG homeostasis. Through functional characterization of recombinant AcvB variants, we showed that the C-terminal domain of AcvB (residues 232-456) is sufficient for full enzymatic activity and defined key residues for catalysis. Absence of the hydrolase resulted in ~10-fold increase in L-PG in Agrobacterium membranes and abolished T-DNA transfer and tumor formation. Overproduction of the L-PG synthase gene (lpiA) in wild-type A. tumefaciens resulted in a similar increase in the L-PG content (~7-fold) and a virulence defect even in the presence of intact AcvB. These results suggest that elevated L-PG amounts (either by overproduction of the synthase or absence of the hydrolase) are responsible for the virulence phenotype. Gradually increasing the L-PG content by complementation with different acvB variants revealed that cellular L-PG levels above 3% of total phospholipids interfere with T-DNA transfer. Cumulatively, this study identified AcvB as a novel virulence factor required for membrane lipid homeostasis and T-DNA transfer.
Mol Microbiol. 2019 Jan;111(1):269-286. doi: 10.1111/mmi.14154. Epub 2018 Nov 14.
1365-2958
30353924
10.1111/mmi.14154
http://hdl.handle.net/10033/621727
Virulence of Agrobacterium tumefaciens requires lipid homeostasis mediated by the lysyl-phosphatidylglycerol hydrolase AcvB.
oai:repository.helmholtz-hzi.de:10033/6217312019-08-30T11:33:04Zcom_10033_128109com_10033_311624com_10033_6839col_10033_128110col_10033_311625
00925njm 22002777a 4500
dc
Łyszkiewicz, Marcin
author
Winter, Samantha J
author
Witzlau, Katrin
author
Föhse, Lisa
author
Brownlie, Rebecca
author
Puchałka, Jacek
author
Verheyden, Nikita A
author
Kunze-Schumacher, Heike
author
Imelmann, Esther
author
Blume, Jonas
author
Raha, Solaiman
author
Sekiya, Takashi
author
Yoshimura, Akihiko
author
Frueh, Jochen T
author
Ullrich, Evelyn
author
Huehn, Jochen
author
Weiss, Siegfried
author
Gutierrez, Maximiliano G
author
Prinz, Immo
author
Zamoyska, Rose
author
Ziętara, Natalia
author
Krueger, Andreas
author
2019-03-01
The interdependence of selective cues during development of regulatory T cells (Treg cells) in the thymus and their suppressive function remains incompletely understood. Here, we analyzed this interdependence by taking advantage of highly dynamic changes in expression of microRNA 181 family members miR-181a-1 and miR-181b-1 (miR-181a/b-1) during late T-cell development with very high levels of expression during thymocyte selection, followed by massive down-regulation in the periphery. Loss of miR-181a/b-1 resulted in inefficient de novo generation of Treg cells in the thymus but simultaneously permitted homeostatic expansion in the periphery in the absence of competition. Modulation of T-cell receptor (TCR) signal strength in vivo indicated that miR-181a/b-1 controlled Treg-cell formation via establishing adequate signaling thresholds. Unexpectedly, miR-181a/b-1-deficient Treg cells displayed elevated suppressive capacity in vivo, in line with elevated levels of cytotoxic T-lymphocyte-associated 4 (CTLA-4) protein, but not mRNA, in thymic and peripheral Treg cells. Therefore, we propose that intrathymic miR-181a/b-1 controls development of Treg cells and imposes a developmental legacy on their peripheral function.
1545-7885
30856173
10.1371/journal.pbio.2006716
http://hdl.handle.net/10033/621731
PLOS Biology
miR-181a/b-1 controls thymic selection of Treg cells and tunes their suppressive capacity.
oai:repository.helmholtz-hzi.de:10033/6217332019-08-30T11:33:05Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Halbedel, Sven
author
Prager, Rita
author
Banerji, Sangeeta
author
Kleta, Sylvia
author
Trost, Eva
author
Nishanth, Gopala
author
Alles, Georg
author
Hölzel, Christina
author
Schlesiger, Friederike
author
Pietzka, Ariane
author
Schlüter, Dirk
author
Flieger, Antje
author
2019-01-01
An outbreak with a remarkable Listeria monocytogenes clone causing 163 cases of non-invasive listeriosis occurred in Germany in 2015. Core genome multi locus sequence typing grouped non-invasive outbreak isolates and isolates obtained from related food samples into a single cluster, but clearly separated genetically close isolates obtained from invasive listeriosis cases. A comparative genomic approach identified a premature stop codon in the chiB gene, encoding one of the two L. monocytogenes chitinases, which clustered with disease outcome. Correction of this premature stop codon in one representative gastroenteritis outbreak isolate restored chitinase production, but effects in infection experiments were not found. While the exact role of chitinases in virulence of L. monocytogenes is still not fully understood, our results now clearly show that ChiB-derived activity is not required to establish L. monocytogenes gastroenteritis in humans. This limits a possible role of ChiB in human listeriosis to later steps of the infection.
2222-1751
30866756
10.1080/22221751.2018.1558960
http://hdl.handle.net/10033/621733
Emerging microbes & infections
Molecular surveillance
chitin
core genome MLST
invasion
listeriosis
A Listeria monocytogenes ST2 clone lacking chitinase ChiB from an outbreak of non-invasive gastroenteritis.
oai:repository.helmholtz-hzi.de:10033/6218252019-08-30T11:24:27Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Marcellin, Patrick
author
Wong, Dave
author
Sievert, William
author
Buggisch, Peter
author
Petersen, Jörg
author
Flisiak, Robert
author
Manns, Michael
author
Kaita, Kelly
author
Krastev, Zahari
author
Lee, Samuel S
author
Cathcart, Andrea L
author
Crans, Gerald
author
Op den Brouw, Marjoleine
author
Jump, Belinda
author
Gaggar, Anuj
author
Flaherty, John
author
Buti-Ferret, Maria
author
2019-05-28
Background & Aims
Tenofovir disoproxil fumarate (TDF) is a first‐line treatment for chronic hepatitis B (CHB). We aimed to describe the efficacy and safety profiles of TDF treatment for up to 10 years in a well‐described cohort of CHB patients.
Methods
Hepatitis B e antigen (HBeAg)‐negative and HBeAg‐positive patients from two randomised, double‐blind trials (ClinicalTrials. gov: NCT00117676 and NCT00116805) completed 48 weeks of randomised treatment with TDF or adefovir dipivoxil. A subset of these patients was then eligible to receive open‐label TDF treatment for up to 10 years. At Year 10, patients were assessed for virological suppression, alanine aminotransferase (ALT) normalisation, serological response, safety, and tolerability.
Results
Of 641 randomised and treated patients, 585 (91%) entered the open‐label extension phase with 203 (32%) patients completing Year 10 of the study. At Year 10, 118/118 (100%) of HBeAg‐negative patients and 78/80 (98%) of HBeAg‐positive patients with available data achieved hepatitis B virus (HBV) DNA <69 IU/mL, while 88/106 (83%) and 60/77 (78%) patients achieved ALT normalisation, respectively. Of the 23 patients with HBeAg status available at Year 10, 12 (52%) and six (27%) experienced HBeAg loss and seroconversion, respectively. No resistance to TDF was documented up to Year 10. In the period between Year 8 and Year 10, the safety profile of TDF was similar to previous reports, with few patients experiencing renal‐ or bone‐related adverse events.
Conclusions
Over 10 years, TDF had a favourable safety profile, was well tolerated, and resulted in continued maintenance of virological suppression with no documented resistance.
Liver Int. 2019 May 28. doi: 10.1111/liv.14155.
1478-3231
31136052
10.1111/liv.14155
http://hdl.handle.net/10033/621825
Liver international
TDF
hepatitis B
long-term
Ten-year efficacy and safety of tenofovir disoproxil fumarate treatment for chronic hepatitis B virus infection.
oai:repository.helmholtz-hzi.de:10033/6218882019-08-30T11:26:40Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Ratner, Hannah K
author
Escalera-Maurer, Andrés
author
Le Rhun, Anaïs
author
Jaggavarapu, Siddharth
author
Wozniak, Jessie E
author
Crispell, Emily K
author
Charpentier, Emmanuelle
author
Weiss, David S
author
2019-06-24
In addition to defense against foreign DNA, the CRISPR-Cas9 system of Francisella novicida represses expression of an endogenous immunostimulatory lipoprotein. We investigated the specificity and molecular mechanism of this regulation, demonstrating that Cas9 controls a highly specific regulon of four genes that must be repressed for bacterial virulence. Regulation occurs through a protospacer adjacent motif (PAM)-dependent interaction of Cas9 with its endogenous DNA targets, dependent on a non-canonical small RNA (scaRNA) and tracrRNA. The limited complementarity between scaRNA and the endogenous DNA targets precludes cleavage, highlighting the evolution of scaRNA to repress transcription without lethally targeting the chromosome. We show that scaRNA can be reprogrammed to repress other genes, and with engineered, extended complementarity to an exogenous target, the repurposed scaRNA:tracrRNA-FnoCas9 machinery can also direct DNA cleavage. Natural Cas9 transcriptional interference likely represents a broad paradigm of regulatory functionality, which is potentially critical to the physiology of numerous Cas9-encoding pathogenic and commensal organisms.
Mol Cell. 2019 Jun 24. pii: S1097-2765(19)30401-0. doi: 10.1016/j.molcel.2019.05.029.
1097-4164
31256988
10.1016/j.molcel.2019.05.029
http://hdl.handle.net/10033/621888
Molecular Cell
CRISPR
Cas9
Francisella novicida
bacterial pathogenesis
gene regulation
transcription
Catalytically Active Cas9 Mediates Transcriptional Interference to Facilitate Bacterial Virulence.
oai:repository.helmholtz-hzi.de:10033/6219032019-08-30T11:26:11Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Pesarrodona, Mireia
author
Jauset, Toni
author
Díaz-Riascos, Zamira V.
author
Sánchez-Chardi, Alejandro
author
Beaulieu, Marie Eve
author
Seras-Franzoso, Joaquin
author
Sánchez-García, Laura
author
Baltà-Foix, Ricardo
author
Mancilla, Sandra
author
Fernández, Yolanda
author
Rinas, Ursula
author
Schwartz, Simó
author
Soucek, Laura
author
Villaverde, Antonio
author
Abasolo, Ibane
author
Vázquez, Esther
author
2019-01-01
Two structurally and functionally unrelated proteins, namely Omomyc and p31, are engineered as CD44-targeted inclusion bodies produced in recombinant bacteria. In this unusual particulate form, both types of protein materials selectively penetrate and kill CD44+ tumor cells in culture, and upon local administration, promote destruction of tumoral tissue in orthotropic mouse models of human breast cancer. These findings support the concept of bacterial inclusion bodies as versatile protein materials suitable for application in chronic diseases that, like cancer, can benefit from a local slow release of therapeutic proteins
10.1002/advs.201900849
https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85069946471&origin=inward
http://hdl.handle.net/10033/621903
Advanced Science
2-s2.0-85069946471
SCOPUS_ID:85069946471
biofabrication
cancer therapy
functional amyloids
inclusion bodies
protein drug release
Targeting Antitumoral Proteins to Breast Cancer by Local Administration of Functional Inclusion Bodies
oai:repository.helmholtz-hzi.de:10033/6219822019-10-18T02:44:02Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Soon, Chai Fen
author
Zhang, Shihong
author
Suneetha, Pothakamuri Venkata
author
Antunes, Dinler Amaral
author
Manns, Michael Peter
author
Raha, Solaiman
author
Schultze-Florey, Christian
author
Prinz, Immo
author
Wedemeyer, Heiner
author
Sällberg Chen, Margaret
author
Cornberg, Markus
author
2019-01-01
T cell immunotherapy is a concept developed for the treatment of cancer and infectious diseases, based on cytotoxic T lymphocytes to target tumor- or pathogen-specific antigens. Antigen-specificity of the T cell receptors (TCRs) is an important selection criterion in the developmental design of immunotherapy. However, off-target specificity is a possible autoimmunity concern if the engineered antigen-specific T cells are cross-reacting to self-peptides in-vivo. In our recent work, we identified several hepatitis E virus (HEV)-specific TCRs as potential candidates to be developed into T cell therapy to treat chronic hepatitis E. One of the identified TCRs, targeting a HLA-A2-restricted epitope at the RNA-dependent RNA polymerase (HEV-1527: LLWNTVWNM), possessed a unique multiple glycine motif in the TCR-β CDR3, which might be a factor inducing cross-reactivity. The aim of our study was to explore if this TCR could cross-recognize self-peptides to underlay autoimmunity. Indeed, we found that this HEV-1527-specific TCR could also cross-recognize an apoptosis-related epitope, Nonmuscle Myosin Heavy Chain 9 (MYH9-478: QLFNHTMFI). While this TCR had dual specificities to both viral epitope and a self-antigen by double Dextramer binding, it was selectively functional against HEV-1527 but not activated against MYH9-478. The consecutive glycine motif in β chain may be the reason promoting TCR binding promiscuity to recognize a secondary target, thereby facilitating cross-recognition. In conclusion, candidate TCRs for immunotherapy development should be screened for autoimmune potential, especially when the TCRs exhibit unique sequence pattern.
Front Immunol. 2019 Sep 4;10:2076. doi: 10.3389/fimmu.2019.02076. eCollection 2019.
1664-3224
31552033
10.3389/fimmu.2019.02076
http://hdl.handle.net/10033/621982
Frontiers in Immunology
CD8+ T cells
T cell receptor (TCR)
T cell therapy
TCR redirection
cross-reactivity
hepatitis E virus (HEV)
immunotherapy
Hepatitis E Virus (HEV)-Specific T Cell Receptor Cross-Recognition: Implications for Immunotherapy.
oai:repository.helmholtz-hzi.de:10033/6219862019-10-19T01:32:20Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Wiegand, Steffen B.
author
Beggel, Bastian
author
Wranke, Anika
author
Aliabadi, Elmira
author
Jaroszewicz, Jerzy
author
Xu, Cheng Jian
author
Li, Yang
author
Manns, Michael P.
author
Lengauer, Thomas
author
Wedemeyer, Heiner
author
Kraft, Anke R.M.
author
Falk, Christine S.
author
Cornberg, Markus
author
2019-10-01
Chronic hepatitis B virus (HBV) infection may follow four different consecutive phases, which are defined by virology as well as biochemical markers and differ in terms of prognosis and need for antiviral treatment. Currently, host responses reflected by immune markers are not considered in this definition. We aimed to study soluble immune markers and their distribution in different phases of chronic HBV infection. In this cross-sectional retrospective study, we investigated a panel of 14 soluble immune markers (SIM) including CXCL10 in 333 patients with chronic HBV infection. In a small cohort of HBeAg positive patients we analyzed SIM before and after HBeAg seroconversion and compared seroconverters to patients with unknown outcome. Significant differences were documented in the levels of several SIM between the four phases of chronic HBV infection. The most pronounced difference among all investigated SIM was observed for CXCL10 concentrations with highest levels in patients with hepatitis. TGF-β and IL-17 revealed different levels between HBeAg negative patients. HBeAg positive patients with HBeAg seroconversion presented higher amounts of IL-12 before seroconversion compared to HBeAg positive patients with unknown follow up. SIM such as CXCL10 but also IL-12, TGF-β and IL-17 may be useful markers to further characterize the phase of chronic HBV infection.
Sci Rep. 2019 Oct 1;9(1):14118. doi: 10.1038/s41598-019-50729-5.
31575964
10.1038/s41598-019-50729-5
https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85072848113&origin=inward
http://hdl.handle.net/10033/621986
Scientific reports
2-s2.0-85072848113
SCOPUS_ID:85072848113
Soluble immune markers in the different phases of chronic hepatitis B virus infection
oai:repository.helmholtz-hzi.de:10033/6219992019-11-02T05:10:58Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Mahalunkar, Sneha
author
Yadav, Amit Singh
author
Gorain, Mahadeo
author
Pawar, Vinay
author
Braathen, Ranveig
author
Weiss, Siegfried
author
Bogen, Bjarne
author
Gosavi, Suresh W.
author
Kundu, Gopal C.
author
2019-01-01
Background: Curcumin has been widely used owing to its various medicinal properties including antitumor effects. However, its clinical application is limited by its instability, poor solubility and low bioavailability. Folic acid (FA)-functionalized nanoformulations may enhance the sustained release of an anticancer drug (curcumin) by tumor-specific targeting to improve therapeutic benefit. This study aims to design a nanoconjugate (NC) comprised of folate–curcumin-loaded gold–polyvinylpyrrolidone nanoparticles (FA–CurAu-PVP NPs) for targeted delivery in breast cancer model systems. Methods: We developed curcumin-loaded FA-functionalized Au-PVP NCs by layer-by-layer assembly. The folic acid–curcumin Au-PVP NCs (FA–CurAu-PVP NCs) were characterized by ultraviolet–visible spectra, Fourier transform infrared spectroscopy, X-ray powder diffraction and thermogravimetric analysis. In vitro anticancer and antimigratory effects of NCs were examined by performing MTT and wound migration assays. The in vivo antitumor efficacy of NCs was investigated using a preclinical breast cancer orthotopic mouse model. Results: Curcumin (40 µg/mL) was loaded along with conjugation of folate onto Au-PVP NPs to form FA–CurAu-PVP NCs. The size and charge of the NCs were increased gradually through layer-by-layer assembly and showed 80% release of curcumin at acidic pH. The NC did not show aggregation when incubated with human serum and mimicked an intrinsic peroxidase-like property in the presence of 3,3ʹ,5,5ʹ-tetramethylbenzidine substrate. The MTT data using these NCs showed efficient anticancer activity at lower doses in estrogen/ progesterone receptor (ER/PR)-negative cells compared with ER/PR-positive cells. Furthermore, the NCs did not show cytotoxicity at the investigated concentration in human breast epithelial and mouse fibroblast cell lines. They showed inhibitory effects on cell migration and high antitumor efficacy in in vivo analysis. Conclusion: These results suggest that folate-based tumor targeting using CurAu-PVP NCs is a promising approach for tumor-specific therapy of breast cancer without harming normal cells.
11769114
10.2147/IJN.S215142
https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85073769401&origin=inward
http://hdl.handle.net/10033/621999
2-s2.0-85073769401
SCOPUS_ID:85073769401
Breast cancer cell line
Curcumin
Cytotoxic activity
Folic acid
Gold nanoconjugate
Functional design of pH-responsive folate-targeted polymer-coated gold nanoparticles for drug delivery and in vivo therapy in breast cancer
oai:repository.helmholtz-hzi.de:10033/6220182019-11-16T02:20:41Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Höfer, Anne
author
Jonigk, Danny
author
Hartleben, Björn
author
Verboom, Murielle
author
Hallensleben, Michael
author
Hübscher, Stefan G
author
Manns, Michael P
author
Jaeckel, Elmar
author
Taubert, Richard
author
2019-10-23
Background:
Subclinical T cell-mediated rejection (subTCMR) is commonly found after liver transplantation and has a good short-term prognosis, even when it is left untreated. Donor-specific antibodies (DSA) are putatively associated with a worse prognosis for recipient and graft after liver transplantation.
Methods:
To assess the immune regulation in subTCMR grafts, gene expression of 93 transcripts for graft injury, tolerance and immune regulation was analyzed in 77 biopsies with “no histological rejection” (NHR; n=25), “clinical TCMR” (cTMCR; n=16) and subTCMR (n=36). In addition, all available subTCMR biopsies (n=71) were tested for DSA with bead assays.
Results:
SubTCMR showed heterogeneous and intermediate expression profiles of transcripts that were upregulated in cTCMR. Graft gene expression suggested a lower activation of effector lymphocytes and a higher activation of regulatory T cells in grafts with subTCMR compared to cTCMR.DSA positivity in subTCMR was associated with histological evidence of more severe graft inflammation and fibrosis. This more severe DSA+ associated graft injury in subTCMR was converged with an upregulation of cTCMR associated transcripts. In nonsupervised analysis DSA positive subTCMR mostly clustered together with cTCMR, while DSA negative subTCMR clustered together with NHR.
Conclusion:
T cell-mediated rejection seem to form a continuum of alloimmune activation. Although subTCMR exhibited less expression of TCMR associated transcript, DSA positivity in subTCMR was associated with an upregulation of rejection associated transcripts. The identification of DSA positive subclinical rejection might help to define patients with more inflammation in the graft and development of fibrosis.
Transplantation. 2019 Oct 23. doi: 10.1097/TP.0000000000003034.
1534-6080
31651790
10.1097/TP.0000000000003034
http://hdl.handle.net/10033/622018
Transplantation
DSA are associated with more graft injury, more fibrosis and upregulation of rejection associated transcripts in subclinical rejection.
oai:repository.helmholtz-hzi.de:10033/6220202019-11-21T02:07:04Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Schlicker, Lisa
author
Szebenyi, Doletha M E
author
Ortiz, Semira R
author
Heinz, Alexander
author
Hiller, Karsten
author
Field, Martha S
author
2019-11-01
The low-calorie sweetener erythritol is endogenously produced from glucose through the pentose phosphate pathway in humans. Erythritol is of medical interest because elevated plasma levels of this polyol are predictive for visceral adiposity gain and development of type 2 diabetes. However, the mechanisms behind these associations remain unknown because the erythritol biosynthesis pathway, particularly the enzyme catalyzing the final step of erythritol synthesis (reduction of erythrose to erythritol), is not characterized. In this study, we purified two enzymes from rabbit liver capable of catalyzing the conversion of erythrose to erythritol: alcohol dehydrogenase 1 (ADH1) and sorbitol dehydrogenase (SORD). Both recombinant human ADH1 and SORD reduce erythrose to erythritol, using NADPH as a co-factor, and cell culture studies indicate that this activity is primarily NADPH-dependent. We found that ADH1 variants vary markedly in both their affinity for erythrose and their catalytic capacity (turnover number). Interestingly, the recombinant protein produced from the ADH1B2 variant, common in Asian populations, is not active when NADPH is used as a co-factor in vitro We also confirmed SORD contributes to intracellular erythritol production in human A549 lung cancer cells, where ADH1 is minimally expressed. In summary, human ADH1 and SORD catalyze the conversion of erythrose to erythritol, pointing to novel roles for two dehydrogenase proteins in human glucose metabolism that may contribute to individual responses to diet. Proteomics data are available via ProteomeXchange with identifier PXD015178.
J Biol Chem. 2019 Nov 1;294(44):16095-16108. doi: 10.1074/jbc.RA119.009049. Epub 2019 Sep 11.
1083-351X
31511322
10.1074/jbc.RA119.009049
http://hdl.handle.net/10033/622020
Journal of Biological Chemistry
alcohol dehydrogenase (ADH)
biomarker
enzyme catalysis
enzyme kinetics
erythritol
glucose metabolism
sorbitol dehydrogenase
Unexpected roles for ADH1 and SORD in catalyzing the final step of erythritol biosynthesis.
oai:repository.helmholtz-hzi.de:10033/6220242019-11-22T02:02:02Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Rinker, Franziska
author
Bremer, Corinna M
author
Schröder, Kathrin
author
Wiegand, Steffen B
author
Bremer, Birgit
author
Manns, Michael P
author
Kraft, Anke R
author
Wedemeyer, Heiner
author
Yang, Lei
author
Pavlovic, Vedran
author
Wat, Cynthia
author
Gerlich, Wolfram H
author
Glebe, Dieter
author
Cornberg, Markus
author
2019-11-13
BACKGROUND & AIMS: Hepatitis B virus (HBV) contains three viral surface proteins, large, middle and small hepatitis B surface protein (LHBs, MHBs, SHBs). Proportions of LHBs and MHBs are lower in patients with inactive versus active chronic infection. Interferon alfa may convert HBeAg-positive chronic hepatitis B (CHB) to an inactive carrier state, but prediction of sustained response is unsatisfactory. The aim of this study was to test the hypothesis that quantification of MHBs and LHBs may allow for a better prognosis of therapeutic response than total hepatitis B surface antigen (HBsAg) concentration.
METHODS: HBs proteins were measured before and during peginterferon alfa-2a therapy in serum from 127 Asian patients with HBeAg-positive CHB. Sustained response was defined as hepatitis B e antigen (HBeAg) seroconversion 24 weeks post-treatment.
RESULTS: Mean total HBs levels were significantly lower in responders versus nonresponders at all time points (P<.05) and decreased steadily during the initial 24 weeks' treatment (by 1.16 versus 0.86 ng/mL in responders/nonresponders, respectively) with unchanged relative proportions. Genotype B had a twofold higher proportion of LHBs than genotype C (13% versus 6%). HBV DNA, HBeAg, HBsAg, and HBs protein levels predicted response equally well but not optimally (area under the ROC curve values >0.70).
CONCLUSIONS: HBs proteins levels differ by HBV genotype. However, quantification of HBs proteins has no advantage over the already established HBsAg assays to predict response to peginterferon alfa-2a therapy in HBeAg-positive patients.
Liver Int. 2019 Nov 13. doi: 10.1111/liv.14298.
1478-3231
31721419
10.1111/liv.14298
http://hdl.handle.net/10033/622024
Liver international
HBs proteins
HBsAg
peginterferon alfa-2a
predictors of response
subviral particles
Quantitation of large, middle and small hepatitis B surface proteins in HBeAg-positive patients treated with peginterferon alfa-2a.
oai:repository.helmholtz-hzi.de:10033/6220472019-12-12T02:00:13Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Bolten, Svenja Nicolin
author
Rinas, Ursula
author
Scheper, Thomas
author
2018-10-01
Heparin is a highly sulfated polysaccharide which belongs to the family of glycosaminoglycans. It is involved in various important biological activities. The major biological purpose is the inhibition of the coagulation cascade to maintain the blood flow in the vasculature. These properties are employed in several therapeutic drugs. Heparin's activities are associated with its interaction to various proteins. To date, the structural heparin-protein interactions are not completely understood. This review gives a general overview of specific patterns and functional groups which are involved in the heparin-protein binding. An understanding of the heparin-protein interactions at the molecular level is not only advantageous in the therapeutic application but also in biotechnological application of heparin for downstreaming. This review focuses on the heparin affinity chromatography. Diverse recombinant proteins can be successfully purified by this method. While effective, it is disadvantageous that heparin is an animal-derived material. Animal-based components carry the risk of contamination. Therefore, they are liable to strict quality controls and the validation of effective good manufacturing practice (GMP) implementation. Hence, adequate alternatives to animal-derived components are needed. This review examines strategies to avoid these disadvantages. Thereby, alternatives for the provision of heparin such as chemical synthesized heparin, chemoenzymatic heparin, and bioengineered heparin are discussed. Moreover, the usage of other chromatographic systems mimetic the heparin effect is reviewed.
Appl Microbiol Biotechnol. 2018 Oct;102(20):8647-8660. doi: 10.1007/s00253-018-9263-3. Epub 2018 Aug 9.
1432-0614
30094590
10.1007/s00253-018-9263-3
http://hdl.handle.net/10033/622047
Applied Microbiology and Biotechnology
Animal-component free
Heparin
Heparin affinity chromatography
Heparin-binding proteins
Heparin-protein interactions
Heparin: role in protein purification and substitution with animal-component free material.
oai:repository.helmholtz-hzi.de:10033/6221062020-01-30T04:10:47Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Quaas, Bastian
author
Burmeister, Laura
author
Li, Zhaopeng
author
Satalov, Alexandra
author
Behrens, Peter
author
Hoffmann, Andrea
author
Rinas, Ursula
author
2019-11-20
PURPOSE: There is a plethora of studies on recombinant human bone morphogenetic protein-2 (rhBMP-2) application and delivery systems, but surprisingly few reports address the biophysical properties of the protein which are of crucial importance to develop effective delivery systems or to solve general problems related to rhBMP-2 production, purification, analysis and application.
METHODS:The solubility, stability and bioactivity of rhBMP-2 obtained by renaturation of E. coli derived inclusion bodies was assessed at different pH and in different buffer systems using (dynamic) light scattering and thermal shift assays as well as intrinsic fluorescence measurements and luciferase based bioassays.
RESULTS:
rhBMP-2 is poorly soluble at physiological pH and higher. The presence of divalent anions further decreases the solubility even under acidic conditions. Thermal stability analyses revealed that rhBMP-2 precipitates are more stable compared to the soluble protein. Moreover, correctly folded rhBMP-2 is also bioactive as precipitated protein and precipitates readily dissolve under appropriate buffer conditions. Once properly formed rhBMP-2 also retains biological activity after temporary exposure to high concentrations of chaotropic denaturants. However, care should be taken to discriminate bioactive rhBMP-2 precipitates from misfolded rhBMP-2 aggregates, e.g. resolvability in MES buffer (pH 5) and a discrete peak in thermoshift experiments are mandatory for correctly folded rhBMP-2.
CONCLUSIONS: Our analysis revealed that E. coli derived rhBMP-2 precipitates are not only bioactive but are also more stable compared to the soluble dimeric molecules. Knowledge about these unusual properties will be helpful to design improved delivery systems requiring lower amounts of rhBMP-2 in clinical applications.
Pharm Res. 2019 Nov 20;36(12):184. doi: 10.1007/s11095-019-2705-5.
1573-904X
31748894
10.1007/s11095-019-2705-5
http://hdl.handle.net/10033/622106
Pharmaceutical Research
protein aggregation
protein solubility
protein stability
recombinant human bone morphogenetic protein-2
refolding
Stability and Biological Activity of E. coli Derived Soluble and Precipitated Bone Morphogenetic Protein-2.
oai:repository.helmholtz-hzi.de:10033/6221492020-02-21T02:04:29Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Than, Nwe Ni
author
Hodson, James
author
Schmidt-Martin, Daniel
author
Taubert, Richard
author
Wawman, Rebecca E
author
Botter, Meemee
author
Gautam, Nishant
author
Bock, Kilian
author
Jones, Rebecca
author
Appanna, Gautham D
author
Godkin, Andrew
author
Montano-Loza, Aldo J
author
Lammert, Frank
author
Schramm, Christoph
author
Manns, Michael P
author
Swain, Mark
author
Burak, Kelly W
author
Adams, David H
author
Hirschfield, Gideon M
author
Oo, Ye Htun
author
2019-12-01
Twenty-two patients with type-1 AIH were included, with a median age of 40 years at diagnosis (range 19-79); 15/22 (68%) were female and 18/22 (82%) were Caucasian. The median period from diagnosis to the end of follow-up in these patients was 11 years (range 3-28). Values of alanine aminotransferase, aspartate aminotransferase and albumin improved significantly following rituximab therapy, and were sustained for up to 2 years (all p ≪0.001). Prednisolone doses were significantly reduced by 12 months post-treatment (p = 0.003), with 13/21 (62%) patients having a dose reduction. Over a median post-treatment follow-up period of 6 years (range 1-10), 5 patients developed AIH flares at a median of 22 months post-treatment, giving an estimated 71% freedom from AIH flare at 2 years. Four of these patients received a second course of treatment, of whom 2 had subsequent further flares. No serious adverse events attributable to rituximab were recorded.
JHEP Rep. 2019 Nov 5;1(6):437-445. doi: 10.1016/j.jhepr.2019.10.005. eCollection 2019 Dec.
2589-5559
32039395
10.1016/j.jhepr.2019.10.005
http://hdl.handle.net/10033/622149
JHEP Reports
Autoimmune hepatitis
B cell depletion therapy
Difficult-to-manage
Prednisolone
Rituximab
Efficacy of rituximab in difficult-to-manage autoimmune hepatitis: Results from the International Autoimmune Hepatitis Group.
oai:repository.helmholtz-hzi.de:10033/6221792020-03-06T04:09:26Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Niehaus, Christian E
author
Strunz, Benedikt
author
Cornillet, Martin
author
Falk, Christine S
author
Schnieders, Ansgar
author
Maasoumy, Benjamin
author
Hardtke, Svenja
author
Manns, Michael P
author
Rm Kraft, Anke
author
Björkström, Niklas K
author
Cornberg, Markus
author
2020-02-03
Patients with advanced liver cirrhosis have an increased susceptibility to infections. As part of the cirrhosis-associated immune dysfunction, mucosal associated invariant T (MAIT) cells, that have the capacity to respond towards bacteria, are severely diminished in circulation and liver tissue. However, MAIT cell presence and function in the peritoneal cavity, a common anatomical site for infections in cirrhosis, remain elusive. To study this, matched peripheral blood and ascites fluid were collected from 35 patients with decompensated cirrhosis, with or without spontaneous bacterial peritonitis (SBP). MAIT cell phenotype and function were analyzed using high-dimensional flow cytometry and obtained data was compared to blood samples of healthy controls (n=24) and patients with compensated cirrhosis (n=11). We found circulating MAIT cells to be severely decreased in cirrhotic patients as compared to controls. In contrast, in ascites fluid, MAIT cells were significantly increased together with CD14+ CD16+ monocytes, ILCs, and NK cells. This was paralleled by elevated levels of several pro-inflammatory cytokines and chemokines in ascites fluid as compared to plasma. Peritoneal MAIT cells displayed an activated tissue-resident phenotype and this was corroborated by increased functional responses following stimulation with E. coli or lL-12 + IL-18 as compared to circulating MAIT cells. During SBP, peritoneal MAIT cell frequencies increased most among all major immune cell subsets, suggestive of active homing of MAIT cells to the site of infection. CONCLUSIONS: Despite severely diminished MAIT cell numbers and impaired phenotype in circulation, peritoneal MAIT cells remain abundant, activated, and highly functional in decompensated cirrhosis and are further enriched in SBP. This suggests that peritoneal MAIT cells could be of interest for immune intervention strategies in patients with decompensated liver cirrhosis and SBP.
Hepatology. 2020 Feb 3. doi: 10.1002/hep.31153.
1527-3350
32012321
10.1002/hep.31153
http://hdl.handle.net/10033/622179
Hepathology
Mucosal-associated invariant T cells
advanced liver cirrhosis
bacterial translocation
peritoneal cavity
spontaneous bacterial peritonitis
MAIT cells are enriched and highly functional in ascites of patients with decompensated liver cirrhosis.
oai:repository.helmholtz-hzi.de:10033/6221882020-03-06T04:10:29Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Qi, Cancan
author
Jiang, Yale
author
Yang, Ivana V
author
Forno, Erick
author
Wang, Ting
author
Vonk, Judith M
author
Gehring, Ulrike
author
Smit, Henriëtte A
author
Milanzi, Edith B
author
Carpaij, Orestes A
author
Berg, Marijn
author
Hesse, Laura
author
Brouwer, Sharon
author
Cardwell, Jonathan
author
Vermeulen, Cornelis J
author
Acosta-Pérez, Edna
author
Canino, Glorisa
author
Boutaoui, Nadia
author
van den Berge, Maarten
author
Teichmann, Sarah A
author
Nawijn, Martijn C
author
Chen, Wei
author
Celedón, Juan C
author
Xu, Cheng-Jian
author
Koppelman, Gerard H
author
2020-01-14
1097-6825
31953105
10.1016/j.jaci.2019.12.911
http://hdl.handle.net/10033/622188
Asthma
environmental exposure
epigenetics
rhinitis
united airways
Nasal DNA methylation profiling of asthma and rhinitis.
oai:repository.helmholtz-hzi.de:10033/6222062020-04-24T08:46:56Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Righetti, Francesco
author
Materne, Solange Lise
author
Boss, John
author
Eichner, Hannes
author
Charpentier, Emmanuelle
author
Loh, Edmund
author
2020-02-20
Increasing evidence has demonstrated that regulatory RNA elements such as riboswitches (RS) play a pivotal role in the fine-tuning of bacterial gene expression. In this study, we investigated and characterized a novel transcriptional thiamine pyrophosphate (TPP) RS in the obligate human pathogen N. meningitidis MC58 (serogroup B). This RS is located in the 5´ untranslated region upstream of thiC gene, encoding a protein involved in TPP biosynthesis, an essential cofactor for all living beings. Primer extension revealed the transcriptional start site of thiC. Northern blot analysis of thiC mRNA and reporter gene studies confirmed the presence of an active TPP-sensing RS. Expression patterns of the wild-type RS and site-specific mutants showed that it is an OFF switch that controls transcription elongation of thiC mRNA. Interestingly, the regulatory mechanism of the meningococcal thiC RS resembles the Gram-positive Bacillus subtilis thiC RS rather than the Gram-negative Escherichia coli thiC RS. Therefore, the meningococcal thiC RS represents a rare example of transcriptional RS in a Gram-negative bacterium. We further observed that the RS is actively involved in modulating gene expression in response to different growth media and to supplemented bacterial and eukaryotic cell lysates as possible sources of nutrients in the nasopharynx. Our results suggest that RS-mediated gene regulation could influence meningococcal fitness, through the fine-tuning of biosynthesis and scavenging of nutrients and cofactors, such as thiamine.
RNA Biol. 2020 Feb 20:1-13. doi: 10.1080/15476286.2020.1727188.
32079473
10.1080/15476286.2020.1727188
http://hdl.handle.net/10033/622206
1555-8584
RNA biology
Neisseria meningitidis
RNA
TPP
gene regulation
non-coding RNA
pathogen
regulatory RNA
riboswitch
vitamin
Characterization of a transcriptional TPP riboswitch in the human pathogen Neisseriameningitidis.
oai:repository.helmholtz-hzi.de:10033/6222072020-03-17T02:02:31Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Dudek, Christian-Alexander
author
Schlicker, Lisa
author
Hiller, Karsten
author
Gas chromatography coupled with mass spectrometry can provide an extensive overview of the metabolic state of a biological system. Analysis of raw mass spectrometry data requires powerful data processing software to generate interpretable results. Here we describe a data processing workflow to generate metabolite levels, mass isotopomer distribution, similarity and variability analysis of metabolites in a nontargeted manner, using stable isotope labeling. Using our data analysis software, no bioinformatic or programming background is needed to generate results from raw mass spectrometry data.
Methods Mol Biol. 2020;2088:17-32. doi: 10.1007/978-1-0716-0159-4_2.
31893368
10.1007/978-1-0716-0159-4_2
http://hdl.handle.net/10033/622207
1940-6029
Methods in molecular biology (Clifton, N.J.)
Data analysis
GCMS
Gas chromatography
Mass isotopomer distribution
Mass spectrometry
Metabolism
Nontargeted metabolomics
Stable isotope labeling
Non-Targeted Mass Isotopolome Analysis Using Stable Isotope Patterns to Identify Metabolic Changes.
oai:repository.helmholtz-hzi.de:10033/6222242020-04-08T02:45:20Zcom_10033_311624com_10033_6839com_10033_621723col_10033_621724col_10033_311625
00925njm 22002777a 4500
dc
Osbelt, Lisa
author
Thiemann, Sophie
author
Smit, Nathiana
author
Lesker, Till Robin
author
Schröter, Madita
author
Gálvez, Eric J C
author
Schmidt-Hohagen, Kerstin
author
Pils, Marina C
author
Mühlen, Sabrina
author
Dersch, Petra
author
Hiller, Karsten
author
Schlüter, Dirk
author
Neumann-Schaal, Meina
author
Strowig, Till
author
2020-03-24
The composition of the intestinal microbiota influences the outcome of enteric infections in human and mice. However, the role of specific members and their metabolites contributing to disease severity is largely unknown. Using isogenic mouse lines harboring distinct microbiota communities, we observed highly variable disease kinetics of enteric Citrobacter rodentium colonization after infection. Transfer of communities from susceptible and resistant mice into germ-free mice verified that the varying susceptibilities are determined by microbiota composition. The strongest differences in colonization were observed in the cecum and could be maintained in vitro by coculturing cecal bacteria with C. rodentium. Cohousing of animals as well as the transfer of cultivable bacteria from resistant to susceptible mice led to variable outcomes in the recipient mice. Microbiome analysis revealed that a higher abundance of butyrate-producing bacteria was associated with the resistant phenotype. Quantification of short-chain fatty acid (SCFA) levels before and after infection revealed increased concentrations of acetate, butyrate and propionate in mice with delayed colonization. Addition of physiological concentrations of butyrate, but not of acetate and/or propionate strongly impaired growth of C. rodentium in vitro. In vivo supplementation of susceptible, antibiotic-treated and germ-free mice with butyrate led to the same level of protection, notably only when cecal butyrate concentration reached a concentration higher than 50 nmol/mg indicating a critical threshold for protection. In the recent years, commensal-derived primary and secondary bacterial metabolites emerged as potent modulators of hosts susceptibility to infection. Our results provide evidence that variations in SCFA production in mice fed fibre-rich chow-based diets modulate susceptibility to colonization with Enterobacteriaceae not only in antibiotic-disturbed ecosystems but even in undisturbed microbial communities. These findings emphasise the need for microbiota normalization across laboratory mouse lines for infection experiments with the model-pathogen C. rodentium independent of investigations of diet and antibiotic usage.
PLoS Pathog. 2020 Mar 24;16(3):e1008448. doi: 10.1371/journal.ppat.1008448.
32208465
10.1371/journal.ppat.1008448
http://hdl.handle.net/10033/622224
1553-7374
PLoS pathogens
Variations in microbiota composition of laboratory mice influence Citrobacter rodentium infection via variable short-chain fatty acid production.
oai:repository.helmholtz-hzi.de:10033/6223002020-06-17T01:30:52Zcom_10033_620533com_10033_311624com_10033_6839com_10033_620652com_10033_620618col_10033_620672col_10033_620534col_10033_311625col_10033_620622
00925njm 22002777a 4500
dc
Lipps, Christoph
author
Klein, Franziska
author
Wahlicht, Tom
author
Seiffert, Virginia
author
Butueva, Milada
author
Zauers, Jeannette
author
Truschel, Theresa
author
Luckner, Martin
author
Köster, Mario
author
MacLeod, Roderick
author
Pezoldt, Jörn
author
Hühn, Jochen
author
Yuan, Qinggong
author
Müller, Peter Paul
author
Kempf, Henning
author
Zweigerdt, Robert
author
Dittrich-Breiholz, Oliver
author
Pufe, Thomas
author
Beckmann, Rainer
author
Drescher, Wolf
author
Riancho, Jose
author
Sañudo, Carolina
author
Korff, Thomas
author
Opalka, Bertram
author
Rebmann, Vera
author
Göthert, Joachim R
author
Alves, Paula M
author
Ott, Michael
author
Schucht, Roland
author
Hauser, Hansjörg
author
Wirth, Dagmar
author
May, Tobias
author
2018-03-08
Fundamental research and drug development for personalized medicine necessitates cell cultures from defined genetic backgrounds. However, providing sufficient numbers of authentic cells from individuals poses a challenge. Here, we present a new strategy for rapid cell expansion that overcomes current limitations. Using a small gene library, we expanded primary cells from different tissues, donors, and species. Cell-type-specific regimens that allow the reproducible creation of cell lines were identified. In depth characterization of a series of endothelial and hepatocytic cell lines confirmed phenotypic stability and functionality. Applying this technology enables rapid, efficient, and reliable production of unlimited numbers of personalized cells. As such, these cell systems support mechanistic studies, epidemiological research, and tailored drug development.
Nat Commun. 2018;9(1):994. Published 2018 Mar 8. doi:10.1038/s41467-018-03408-4.
29520052
10.1038/s41467-018-03408-4
http://hdl.handle.net/10033/622300
2041-1723
Nature communications
Expansion of functional personalized cells with specific transgene combinations.
oai:repository.helmholtz-hzi.de:10033/6223572020-07-24T02:28:48Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Bolten, Svenja Nicolin
author
Knoll, Anne-Sophie
author
Li, Zhaopeng
author
Gellermann, Pia
author
Pepelanova, Iliyana
author
Rinas, Ursula
author
Scheper, Thomas
author
2020-08
This paper analyzes the use of animal-component free chromatographic materials for the efficient purifi- cation of the human fibroblast growth factor 2 (hFGF-2). hFGF-2 is produced in Escherichia coli and pu- rified via three different chromatography steps, which include a strong cation exchange chromatography as a capture step, followed by heparin affinity chromatography and an anion exchange chromatography as a polishing step. The affinity chromatography step is based on the animal-derived material heparin. Chemically produced ligands provide a viable alternative to animal-derived components in production processes, since they are characterized by a defined structure which leads to reproducible results and a broad range of applications. The alternative ligands can be assigned to adsorber of the mixed-mode chromatography (MMC) and pseudo-affinity chromatography. Eight different animal-component free materials used as adsorbers in MMC or pseudo-affinity chromatog- raphy were tested as a substitute for heparin. The MMCs were cation exchangers characterized with fur- ther functional residues. The ligands of the pseudo-affinity chromatography were heparin-like ligands which are based on heparin’s molecular structure. The alternative methods were tested as a capture step and in combination with another chromatographic step in the purification procedure of hFGF-2. In each downstream step purity, recovery and yield were analysed and compared to the conventional downstream process. Two types of MMC –the column Foresight TM Nuvia TM cPrime TM from Bio-Rad Laboratories and the col- umn HiTrap TM Capto TM MMC from GE Healthcare Life Sciences - can be regarded as effective animal- component free alternatives to the heparin - based adsorber.
0021-9673
10.1016/j.chroma.2020.461367
http://hdl.handle.net/10033/622357
Journal of Chromatography A
S0021967320306440
Organic Chemistry
Analytical Chemistry
Biochemistry
General Medicine
Purification of the human fibroblast growth factor 2 using novel animal-component free materials
oai:repository.helmholtz-hzi.de:10033/6223662020-07-28T01:25:47Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Sanchez-Ruiz, Monica
author
Brunn, Anna
author
Montesinos-Rongen, Manuel
author
Rudroff, Claudia
author
Hartmann, Melanie
author
Schlüter, Dirk
author
Pfitzer, Gabriele
author
Deckert, Martina
author
2018-12-27
Inflammatory bowel diseases frequently cause gastrointestinal dysmotility, suggesting that they may also affect the enteric nervous system. So far, the precise mechanisms that lead to gastrointestinal dysmotility in inflammatory bowel diseases have not been elucidated. To determine the effect of CD8 T cells on gastrointestinal motility, transgenic mice expressing ovalbumin on enteric neurons were generated. In these mice, adoptive transfer of ovalbumin-specific OT-I CD8 T cells induced severe enteric ganglionitis. CD8 T cells homed to submucosal and myenteric plexus neurons, 60% of which were lost, clinically resulting in severely impaired gastrointestinal transition. Anti-interferon-γ treatment rescued neurons by preventing their up-regulation of major histocompatibility complex class I antigen, thus preserving gut motility. These preclinical murine data translated well into human gastrointestinal dysmotility. In a series of 30 colonic biopsy specimens from patients with gastrointestinal dysmotility, CD8 T cell-mediated ganglionitis was detected that was followed by severe loss of enteric neurons (74.8%). Together, the preclinical and clinical data support the concept that autoimmune CD8 T cells play an important pathogenetic role in gastrointestinal dysmotility and may destroy enteric neurons.
Am J Pathol. 2019;189(3):540-551. doi:10.1016/j.ajpath.2018.11.016.
30593823
10.1016/j.ajpath.2018.11.016
http://hdl.handle.net/10033/622366
1525-2191
The American journal of pathology
Enteric Murine Ganglionitis Induced by Autoimmune CD8 T Cells Mimics Human Gastrointestinal Dysmotility.
oai:repository.helmholtz-hzi.de:10033/6223832020-08-05T01:34:46Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Aguirre-Gamboa, Raúl
author
de Klein, Niek
author
di Tommaso, Jennifer
author
Claringbould, Annique
author
van der Wijst, Monique Gp
author
de Vries, Dylan
author
Brugge, Harm
author
Oelen, Roy
author
Võsa, Urmo
author
Zorro, Maria M
author
Chu, Xiaojin
author
Bakker, Olivier B
author
Borek, Zuzanna
author
Ricaño-Ponce, Isis
author
Deelen, Patrick
author
Xu, Cheng-Jiang
author
Swertz, Morris
author
Jonkers, Iris
author
Withoff, Sebo
author
Joosten, Irma
author
Sanna, Serena
author
Kumar, Vinod
author
Koenen, Hans J P M
author
Joosten, Leo A B
author
Netea, Mihai G
author
Wijmenga, Cisca
author
Franke, Lude
author
Li, Yang
author
2020-06-12
A novel planctomycetal strain, designated Pla85_3_4T, was isolated from the surface of wood incubated at the discharge of a wastewater treatment plant in the Warnow river near Rostock, Germany. Cells of the novel strain have a cell envelope architecture resembling that of Gram-negative bacteria, are round to pear-shaped (length: 2.2 ± 0.4 µm, width: 1.2 ± 0.3 µm), form aggregates and divide by polar budding. Colonies have a cream colour. Strain Pla85_3_4T grows at ranges of 10-30 °C (optimum 26 °C) and at pH 6.5-10.0 (optimum 7.5), and has a doubling time of 26 h. Phylogenetically, strain Pla85_3_4T (DSM 103796T = LMG 29741T) is concluded to represent a novel species of a novel genus within the family Pirellulaceae, for which we propose the name Lignipirellula cremea gen. nov., sp. nov.
BMC Bioinformatics. 2020;21(1):243. Published 2020 Jun 12. doi:10.1186/s12859-020-03576-5.
32532224
10.1186/s12859-020-03576-5
http://hdl.handle.net/10033/622383
1471-2105
BMC bioinformatics
Cell types
Deconvolution
Immune cells
eQTL
Deconvolution of bulk blood eQTL effects into immune cell subpopulations.
oai:repository.helmholtz-hzi.de:10033/6223842020-08-05T01:36:03Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Kurniawan, Henry
author
Franchina, Davide G
author
Guerra, Luana
author
Bonetti, Lynn
author
-Baguet, Leticia Soriano
author
Grusdat, Melanie
author
Schlicker, Lisa
author
Hunewald, Oliver
author
Dostert, Catherine
author
Merz, Myriam P
author
Binsfeld, Carole
author
Duncan, Gordon S
author
Farinelle, Sophie
author
Nonnenmacher, Yannic
author
Haight, Jillian
author
Das Gupta, Dennis
author
Ewen, Anouk
author
Taskesen, Rabia
author
Halder, Rashi
author
Chen, Ying
author
Jäger, Christian
author
Ollert, Markus
author
Wilmes, Paul
author
Vasiliou, Vasilis
author
Harris, Isaac S
author
Knobbe-Thomsen, Christiane B
author
Turner, Jonathan D
author
Mak, Tak W
author
Lohoff, Michael
author
Meiser, Johannes
author
Hiller, Karsten
author
Brenner, Dirk
author
2020-03-25
Regulatory T cells (Tregs) maintain immune homeostasis and prevent autoimmunity. Serine stimulates glutathione (GSH) synthesis and feeds into the one-carbon metabolic network (1CMet) essential for effector T cell (Teff) responses. However, serine's functions, linkage to GSH, and role in stress responses in Tregs are unknown. Here, we show, using mice with Treg-specific ablation of the catalytic subunit of glutamate cysteine ligase (Gclc), that GSH loss in Tregs alters serine import and synthesis and that the integrity of this feedback loop is critical for Treg suppressive capacity. Although Gclc ablation does not impair Treg differentiation, mutant mice exhibit severe autoimmunity and enhanced anti-tumor responses. Gclc-deficient Tregs show increased serine metabolism, mTOR activation, and proliferation but downregulated FoxP3. Limitation of cellular serine in vitro and in vivo restores FoxP3 expression and suppressive capacity of Gclc-deficient Tregs. Our work reveals an unexpected role for GSH in restricting serine availability to preserve Treg functionality.
Cell Metab. 2020;31(5):920-936.e7. doi:10.1016/j.cmet.2020.03.004.
32213345
10.1016/j.cmet.2020.03.004
http://hdl.handle.net/10033/622384
1932-7420
Cell metabolism
FoxP3
ROS
Treg
autoimmunity
cancer
diet
glutamate cysteine ligase
glutathione
one carbon metabolism
serine metabolism
Glutathione Restricts Serine Metabolism to Preserve Regulatory T Cell Function.
oai:repository.helmholtz-hzi.de:10033/6223882020-08-06T02:29:27Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Sundermann, Julius
author
Oehmichen, Sarah
author
Sydow, Steffen
author
Burmeister, Laura
author
Quaas, Bastian
author
Hänsch, Robert
author
Rinas, Ursula
author
Hoffmann, Andrea
author
Menzel, Henning
author
Bunjes, Heike
author
2020-06-30
Polycaprolactone (PCL) fiber mats with different surface modifications were functionalized with a chitosan nanogel coating to attach the growth factor human bone morphogenetic protein 2 (BMP-2). Three different hydrophilic surface modifications were compared with regard to the binding and in vitro release of BMP-2. The type of surface modification and the specific surface area derived from the fiber thickness had an important influence on the degree of protein loading. Coating the PCL fibers with polydopamine resulted in the binding of the largest BMP-2 quantity per surface area. However, most of the binding was irreversible over the investigated period of time, causing a low release in vitro. PCL fiber mats with a chitosan-graft-PCL coating and an additional alginate layer, as well as PCL fiber mats with an air plasma surface modification boundless BMP-2, but the immobilized protein could almost completely be released. With polydopamine and plasma modifications as well as with unmodified PCL, high amounts of BMP-2 could also be attached directly to the surface. Integration of BMP-2 into the chitosan nanogel functionalization considerably increased binding on all hydrophilized surfaces and resulted in a sustained release with an initial burst release of BMP-2 without detectable loss of bioactivity in vitro.
J Biomed Mater Res A. 2020;10.1002/jbm.a.37045. doi:10.1002/jbm.a.37045.
32608183
10.1002/jbm.a.37045
http://hdl.handle.net/10033/622388
1552-4965
Journal of biomedical materials research. Part A
BMP-2
PCL fiber scaffold
chitosan coating
growth factor delivery
surface modification
Varying the sustained release of BMP-2 from chitosan nanogel-functionalized polycaprolactone fiber mats by different polycaprolactone surface modifications.
oai:repository.helmholtz-hzi.de:10033/6223972020-08-12T02:29:46Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Strunz, Benedikt
author
Hengst, Julia
author
Wedemeyer, Heiner
author
Björkström, Niklas K
author
2018-07-25
Diversity is crucial for the immune system to efficiently combat infections. Natural killer (NK) cells are innate cytotoxic lymphocytes that contribute to the control of viral infections. NK cells were for long thought to be a homogeneous population of cells. However, recent work has instead revealed NK cells to represent a highly diverse population of immune cells where a vast number of subpopulations with distinct characteristics exist across tissues. However, the degree to which a chronic viral infection affects NK cell diversity remains elusive. Hepatitis C virus (HCV) is effective in establishing chronic infection in humans. During the last years, new direct-acting antiviral drugs (DAA) have revolutionized treatment of chronic hepatitis C, enabling rapid cure in the majority of patients. This allows us to study the influence of a chronic viral infection and its subsequent elimination on the NK cell compartment with a focus on NK cell diversity. In our recent study (Nat Commun, 9:2275), we show that chronic HCV infection irreversibly impacts human NK cell repertoire diversity.
Cell Stress. 2018;2(8):216-218. Published 2018 Jul 25. doi:10.15698/cst2018.07.150.
31225489
10.15698/cst2018.07.150
http://hdl.handle.net/10033/622397
2523-0204
Cell stress
Hepatitis C virus infection
direct-acting antiviral therapy
natural killer cells
Irreversible impact of chronic hepatitis C virus infection on human natural killer cell diversity.
oai:repository.helmholtz-hzi.de:10033/6224042020-08-14T01:30:08Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Davies, Mark R
author
McIntyre, Liam
author
Mutreja, Ankur
author
Lacey, Jake A
author
Lees, John A
author
Towers, Rebecca J
author
Duchêne, Sebastián
author
Smeesters, Pierre R
author
Frost, Hannah R
author
Price, David J
author
Holden, Matthew T G
author
David, Sophia
author
Giffard, Philip M
author
Worthing, Kate A
author
Seale, Anna C
author
Berkley, James A
author
Harris, Simon R
author
Rivera-Hernandez, Tania
author
Berking, Olga
author
Cork, Amanda J
author
Torres, Rosângela S L A
author
Lithgow, Trevor
author
Strugnell, Richard A
author
Bergmann, Rene
author
Nitsche-Schmitz, Patric
author
Chhatwal, Gusharan S
author
Bentley, Stephen D
author
Fraser, John D
author
Moreland, Nicole J
author
Carapetis, Jonathan R
author
Steer, Andrew C
author
Parkhill, Julian
author
Saul, Allan
author
Williamson, Deborah A
author
Currie, Bart J
author
Tong, Steven Y C
author
Dougan, Gordon
author
Walker, Mark J
author
2019-05-27
Group A Streptococcus (GAS; Streptococcus pyogenes) is a bacterial pathogen for which a commercial vaccine for humans is not available. Employing the advantages of high-throughput DNA sequencing technology to vaccine design, we have analyzed 2,083 globally sampled GAS genomes. The global GAS population structure reveals extensive genomic heterogeneity driven by homologous recombination and overlaid with high levels of accessory gene plasticity. We identified the existence of more than 290 clinically associated genomic phylogroups across 22 countries, highlighting challenges in designing vaccines of global utility. To determine vaccine candidate coverage, we investigated all of the previously described GAS candidate antigens for gene carriage and gene sequence heterogeneity. Only 15 of 28 vaccine antigen candidates were found to have both low naturally occurring sequence variation and high (>99%) coverage across this diverse GAS population. This technological platform for vaccine coverage determination is equally applicable to prospective GAS vaccine antigens identified in future studies.
Nat Genet. 2019;51(6):1035-1043. doi:10.1038/s41588-019-0417-8.
31133745
10.1038/s41588-019-0417-8
http://hdl.handle.net/10033/622404
1546-1718
Nature genetics
PMC6650292
Atlas of group A streptococcal vaccine candidates compiled using large-scale comparative genomics.
oai:repository.helmholtz-hzi.de:10033/6224112020-08-18T01:29:52Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Wang, Xu
author
Mulas, Floriana
author
Yi, Wenjing
author
Brunn, Anna
author
Nishanth, Gopala
author
Just, Sissy
author
Waisman, Ari
author
Brück, Wolfgang
author
Deckert, Martina
author
Schlüter, Dirk
author
2019-04-03
Astrocytes are critical regulators of neuroinflammation in multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE). Growing evidence indicates that ubiquitination of signaling molecules is an important cell-intrinsic mechanism governing astrocyte function during MS and EAE Here, we identified an upregulation of the deubiquitinase OTU domain, ubiquitin aldehyde binding 1 (OTUB1) in astrocytes during MS and EAE Mice with astrocyte-specific OTUB1 ablation developed more severe EAE due to increased leukocyte accumulation, proinflammatory gene transcription, and demyelination in the spinal cord as compared to control mice. OTUB1-deficient astrocytes were hyperactivated in response to IFN-γ, a fingerprint cytokine of encephalitogenic T cells, and produced more proinflammatory cytokines and chemokines than control astrocytes. Mechanistically, OTUB1 inhibited IFN-γ-induced Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling by K48 deubiquitination and stabilization of the JAK2 inhibitor suppressor of cytokine signaling 1 (SOCS1). Thus, astrocyte-specific OTUB1 is a critical inhibitor of neuroinflammation in CNS autoimmunity.
EMBO J. 2019;38(10):e100947. doi:10.15252/embj.2018100947.
30944096
10.15252/embj.2018100947
http://hdl.handle.net/10033/622411
1460-2075
The EMBO journal
OTUB1
astrocyte
experimental autoimmune encephalomyelitis
multiple sclerosis
neuroinflammation
ubiquitination
OTUB1 inhibits CNS autoimmunity by preventing IFN-γ-induced hyperactivation of astrocytes.
oai:repository.helmholtz-hzi.de:10033/6225012020-10-09T01:33:59Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Li, Zhaopeng
author
Rinas, Ursula
author
2020-09-03
A comparison of the metabolic response of Escherichia coli BL21 (DE3) towards the production of human basic fibroblast growth factor (hFGF-2) or towards carbon overfeeding revealed similarities which point to constraints in anabolic pathways. Contrary to expectations, neither energy generation (e.g., ATP) nor provision of precursor molecules for nucleotides (e.g., uracil) and amino acids (e.g., pyruvate, glutamate) limit host cell and plasmid-encoded functions. Growth inhibition is assumed to occur when hampered anabolic capacities do not match with the ongoing and overwhelming carbon catabolism. Excessive carbon uptake leads to by-product secretion, for example, pyruvate, acetate, glutamate, and energy spillage, for example, accumulation and degradation of adenine nucleotides with concomitant accumulation of extracellular hypoxanthine. The cellular response towards compromised anabolic capacities involves downregulation of cAMP formation, presumably responsible for subsequently better-controlled glucose uptake and resultant accumulation of glucose in the culture medium. Growth inhibition is neglectable under conditions of reduced carbon availability when hampered anabolic capacities also match with catabolic carbon processing. The growth inhibitory effect with accompanying energy spillage, respectively, hypoxanthine secretion and cessation of cAMP formation is not unique to the production of hFGF-2 but observed during the production of other proteins and also during overexpression of genes without transcript translation.
Biotechnol Bioeng. 2020 Sep 3. doi: 10.1002/bit.27553. Epub ahead of print.
32880889
10.1002/bit.27553
http://hdl.handle.net/10033/622501
1097-0290
Biotechnology and bioengineering
Escherichia coli
carbon overfeeding response
energy spilling and hypoxanthine secretion
recombinant protein production-associated metabolic burden
Recombinant protein production-associated metabolic burden reflects anabolic constraints and reveals similarities to a carbon overfeeding response.
oai:repository.helmholtz-hzi.de:10033/6225192020-10-17T01:32:44Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Steinert, Michael
author
Ramming, Isabell
author
Bergmann, Simone
author
2020-09-03
Von Willebrand factor (VWF) is a mechano-sensitive protein with crucial functions in normal hemostasis, which are strongly dependant on the shear-stress mediated defolding and multimerization of VWF in the blood stream. Apart from bleeding disorders, higher plasma levels of VWF are often associated with a higher risk of cardiovascular diseases. Herein, the disease symptoms are attributed to the inflammatory response of the activated endothelium and share high similarities to the reaction of the host vasculature to systemic infections caused by pathogenic bacteria such as Staphylococcus aureus and Streptococcus pneumoniae. The bacteria recruit circulating VWF, and by binding to immobilized VWF on activated endothelial cells in blood flow, they interfere with the physiological functions of VWF, including platelet recruitment and coagulation. Several bacterial VWF binding proteins have been identified and further characterized by biochemical analyses. Moreover, the development of a combination of sophisticated cell culture systems simulating shear stress levels of the blood flow with microscopic visualization also provided valuable insights into the interaction mechanism between bacteria and VWF-strings. In vivo studies using mouse models of bacterial infection and zebrafish larvae provided evidence that the interaction between bacteria and VWF promotes bacterial attachment, coagulation, and thrombus formation, and thereby contributes to the pathophysiology of severe infectious diseases such as infective endocarditis and bacterial sepsis. This mini-review summarizes the current knowledge of the interaction between bacteria and the mechano-responsive VWF, and corresponding pathophysiological disease symptoms.
Front Med (Lausanne). 2020 Sep 3;7:543. doi: 10.3389/fmed.2020.00543.
2296-858X
33015097
10.3389/fmed.2020.00543
http://hdl.handle.net/10033/622519
Frontiers in medicine
Staphylococcus areus
Streptococcus pneumoniae
microfluidic
sepsis
von Willebrand
Impact of Von Willebrand Factor on Bacterial Pathogenesis.
oai:repository.helmholtz-hzi.de:10033/6225482020-11-03T01:31:08Zcom_10033_311624com_10033_6839com_10033_620644col_10033_311625col_10033_620646
00925njm 22002777a 4500
dc
Knittel, Vanessa
author
Sadana, Pooja
author
Seekircher, Stephanie
author
Stolle, Anne-Sophie
author
Körner, Britta
author
Volk, Marcel
author
Jeffries, Cy M
author
Svergun, Dmitri I
author
Heroven, Ann Kathrin
author
Scrima, Andrea
author
Dersch, Petra
author
2020-09-23
Type VI secretion systems (T6SSs) are complex macromolecular injection machines which are widespread in Gram-negative bacteria. They are involved in host-cell interactions and pathogenesis, required to eliminate competing bacteria, or are important for the adaptation to environmental stress conditions. Here we identified regulatory elements controlling the T6SS4 of Yersinia pseudotuberculosis and found a novel type of hexameric transcription factor, RovC. RovC directly interacts with the T6SS4 promoter region and activates T6SS4 transcription alone or in cooperation with the LysR-type regulator RovM. A higher complexity of regulation was achieved by the nutrient-responsive global regulator CsrA, which controls rovC expression on the transcriptional and post-transcriptional level. In summary, our work unveils a central mechanism in which RovC, a novel key activator, orchestrates the expression of the T6SS weapons together with a global regulator to deploy the system in response to the availability of nutrients in the species' native environment.
LoS Pathog. 2020 Sep 23;16(9):e1008552. doi: 10.1371/journal.ppat.1008552.
32966346
10.1371/journal.ppat.1008552
http://hdl.handle.net/10033/622548
1553-7374
PLoS pathogens
RovC - a novel type of hexameric transcriptional activator promoting type VI secretion gene expression.
oai:repository.helmholtz-hzi.de:10033/6225532020-11-04T04:38:08Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Zayed, Ashraf R
author
Pecellin, Marina
author
Salah, Alaa
author
Alalam, Hanna
author
Butmeh, Suha
author
Steinert, Michael
author
Lesnik, Rene
author
Brettar, Ingrid
author
Höfle, Manfred G
author
Bitar, Dina M
author
2020-10-22
The West Bank can be considered a high-risk area for Legionnaires' disease (LD) due to its hot climate, intermittent water supply and roof storage of drinking water. Legionella, mostly L. pneumophila, are responsible for LD, a severe, community-acquired and nosocomial pneumonia. To date, no extensive assessment of Legionella spp and L. pneumophila using cultivation in combination with molecular approaches in the West Bank has been published. Two years of environmental surveillance of Legionella in water and biofilms in the drinking water distribution systems (DWDS) of eight hospitals was carried out; 180 L. pneumophila strains were isolated, mostly from biofilms in DWDS. Most of the isolates were identified as serogroup (Sg) 1 (60%) and 6 (30%), while a minor fraction comprised Sg 8 and 10. Multilocus Variable number of tandem repeats Analysis using 13 loci (MLVA-8(12)) was applied as a high-resolution genotyping method and compared to the standard Sequence Based Typing (SBT). The isolates were genotyped in 27 MLVA-8(12) genotypes (Gt), comprising four MLVA clonal complexes (VACC 1; 2; 5; 11). The major fraction of isolates constituted Sequence Type (ST)1 and ST461. Most of the MLVA-genotypes were highly diverse and often unique. The MLVA-genotype composition showed substantial regional variability. In general, the applied MLVA-method made it possible to reproducibly genotype the isolates, and was consistent with SBT but showed a higher resolution. The advantage of the higher resolution was most evident for the subdivision of the large strain sets of ST1 and ST461; these STs were shown to be highly pneumonia-relevant in a former study. This shows that the resolution by MLVA is advantageous for back-tracking risk sites and for the avoidance of outbreaks of L. pneumophila. Overall, our results provide important insights into the detailed population structure of L. pneumophila, allowing for better risk assessment for DWDS.
Pathogens. 2020 Oct 22;9(11):E862. doi: 10.3390/pathogens9110862.
2076-0817
33105606
10.3390/pathogens9110862
http://hdl.handle.net/10033/622553
Pathogens (Basel, Switzerland)
Legionella pneumophila
MLVA-genotypes
West Bank
clonal complex
hospital water
Characterization of Populations by Multilocus Variable Number of Tandem Repeats (MLVA) Genotyping from Drinking Water and Biofilm in Hospitals from Different Regions of the West Bank.
oai:repository.helmholtz-hzi.de:10033/6225742020-11-12T02:56:44Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Hoang, Thanh T
author
Sikdar, Sinjini
author
Xu, Cheng-Jian
author
Lee, Mi Kyeong
author
Cardwell, Jonathan
author
Forno, Erick
author
Imboden, Medea
author
Jeong, Ayoung
author
Madore, Anne-Marie
author
Qi, Cancan
author
Wang, Tianyuan
author
Bennett, Brian D
author
Ward, James M
author
Parks, Christine G
author
Beane-Freeman, Laura E
author
King, Debra
author
Motsinger-Reif, Alison
author
Umbach, David M
author
Wyss, Annah B
author
Schwartz, David A
author
Celedón, Juan C
author
Laprise, Catherine
author
Ober, Carole
author
Probst-Hensch, Nicole
author
Yang, Ivana V
author
Koppelman, Gerard H
author
London, Stephanie J
author
2020-09-03
Epigenome-wide studies of methylation in children support a role for epigenetic mechanisms in asthma; however, studies in adults are rare and few have examined non-atopic asthma. We conducted the largest epigenome-wide association study (EWAS) of blood DNA methylation in adults in relation to non-atopic and atopic asthma.We measured DNA methylation in blood using the Illumina MethylationEPIC array among 2286 participants in a case-control study of current adult asthma nested within a United States agricultural cohort. Atopy was defined by serum specific immunoglobulin E (IgE). Participants were categorised as atopy without asthma (n=185), non-atopic asthma (n=673), atopic asthma (n=271), or a reference group of neither atopy nor asthma (n=1157). Analyses were conducted using logistic regression.No associations were observed with atopy without asthma. Numerous cytosine-phosphate-guanine (CpG) sites were differentially methylated in non-atopic asthma (eight at family-wise error rate (FWER) p<9×10-8, 524 at false discovery rate (FDR) less than 0.05) and implicated 382 novel genes. More CpG sites were identified in atopic asthma (181 at FWER, 1086 at FDR) and implicated 569 novel genes. 104 FDR CpG sites overlapped. 35% of CpG sites in non-atopic asthma and 91% in atopic asthma replicated in studies of whole blood, eosinophils, airway epithelium, or nasal epithelium. Implicated genes were enriched in pathways related to the nervous system or inflammation.We identified numerous, distinct differentially methylated CpG sites in non-atopic and atopic asthma. Many CpG sites from blood replicated in asthma-relevant tissues. These circulating biomarkers reflect risk and sequelae of disease, as well as implicate novel genes associated with non-atopic and atopic asthma.
Eur Respir J. 2020 Sep 3;56(3):2000217. doi: 10.1183/13993003.00217-2020.
32381493
10.1183/13993003.00217-2020
http://hdl.handle.net/10033/622574
1399-3003
The European respiratory journal
Epigenome-wide association study of DNA methylation and adult asthma in the Agricultural Lung Health Study.
oai:repository.helmholtz-hzi.de:10033/6226042020-11-24T01:42:24Zcom_10033_311624com_10033_6839com_10033_620857com_10033_338554col_10033_621787col_10033_311625col_10033_620858
00925njm 22002777a 4500
dc
Yuyama, Kamila Tomoko
author
Rohde, Manfred
author
Molinari, Gabriella
author
Stadler, Marc
author
Abraham, Wolf-Rainer
author
2020-11-08
Infections involving biofilms are difficult to treat due to increased resistances against antibiotics and the immune system. Hence, there is an urgent demand for novel drugs against biofilm infections. During our search for novel biofilm inhibitors from fungi, we isolated linoleic acid from the ascomycete Hypoxylon fragiforme which showed biofilm inhibition of several bacteria at sub-MIC concentrations. Many fatty acids possess antimicrobial activities, but their minimum inhibitory concentrations (MIC) are high and reports on biofilm interferences are scarce. We demonstrated that not only linoleic acid but several unsaturated long-chain fatty acids inhibited biofilms at sub-MIC concentrations. The antibiofilm activity exerted by long-chain fatty acids was mainly against Gram-positive bacteria, especially against Staphylococcus aureus. Micrographs of treated S. aureus biofilms revealed a reduction in the extracellular polymeric substances, pointing to a possible mode of action of fatty acids on S. aureus biofilms. The fatty acids had a strong species specificity. Poly-unsaturated fatty acids had higher activities than saturated ones, but no obvious rule could be found for the optimal length and desaturation for maximal activity. As free fatty acids are non-toxic and ubiquitous in food, they may offer a novel tool, especially in combination with antibiotics, for the control of biofilm infections.
Antibiotics (Basel). 2020 Nov 8;9(11):E788. doi: 10.3390/antibiotics9110788.
2079-6382
33171584
10.3390/antibiotics9110788
http://hdl.handle.net/10033/622604
Antibiotics (Basel, Switzerland)
Gram-positive bacteria
Hypoxylon fragiforme
Staphylococcus aureus
biofilm inhibition
fatty acid
Unsaturated Fatty Acids Control Biofilm Formation of and Other Gram-Positive Bacteria.
oai:repository.helmholtz-hzi.de:10033/6226052020-11-24T01:42:33Zcom_10033_311624com_10033_6839com_10033_620652col_10033_620673col_10033_620672col_10033_311625
00925njm 22002777a 4500
dc
Gödecke, Natascha
author
Riedel, Jan
author
Herrmann, Sabrina
author
Behme, Sara
author
Rand, Ulfert
author
Kubsch, Tobias
author
Cicin-Sain, Luka
author
Hauser, Hansjörg
author
Köster, Mario
author
Wirth, Dagmar
author
2020-11-18
Mammalian first line of defense against viruses is accomplished by the interferon (IFN) system. Viruses have evolved numerous mechanisms to reduce the IFN action allowing them to invade the host and/or to establish latency. We generated an IFN responsive intracellular hub by integrating the synthetic transactivator tTA into the chromosomal Mx2 locus for IFN-based activation of tTA dependent expression modules. The additional implementation of a synthetic amplifier module with positive feedback even allowed for monitoring and reacting to infections of viruses that can antagonize the IFN system. Low and transient IFN amounts are sufficient to trigger these amplifier cells. This gives rise to higher and sustained-but optionally de-activatable-expression even when the initial stimulus has faded out. Amplification of the IFN response induced by IFN suppressing viruses is sufficient to protect cells from infection. Together, this interfaced sensor/actuator system provides a toolbox for robust sensing and counteracting viral infections.
Nucleic Acids Res. 2020 Nov 18;48(20):11799-11811. doi: 10.1093/nar/gkaa961.
33137201
10.1093/nar/gkaa961
http://hdl.handle.net/10033/622605
1362-4962
Nucleic acids research
Synthetic rewiring and boosting type I interferon responses for visualization and counteracting viral infections.
oai:repository.helmholtz-hzi.de:10033/6226322020-12-04T03:38:49Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Maus, Irena
author
Klocke, Michael
author
Derenkó, Jaqueline
author
Stolze, Yvonne
author
Beckstette, Michael
author
Jost, Carsten
author
Wibberg, Daniel
author
Blom, Jochen
author
Henke, Christian
author
Willenbücher, Katharina
author
Rumming, Madis
author
Rademacher, Antje
author
Pühler, Alfred
author
Sczyrba, Alexander
author
Schlüter, Andreas
author
2020-03-02
Background: Anaerobic digestion (AD) of protein-rich grass silage was performed in experimental two-stage twophase biogas reactor systems at low vs. increased organic loading rates (OLRs) under mesophilic (37 °C) and
thermophilic (55 °C) temperatures. To follow the adaptive response of the biomass-attached cellulolytic/hydrolytic
biofilms at increasing ammonium/ammonia contents, genome-centered metagenomics and transcriptional profiling
based on metagenome assembled genomes (MAGs) were conducted.
Results: In total, 78 bacterial and archaeal MAGs representing the most abundant members of the communities,
and featuring defined quality criteria were selected and characterized in detail. Determination of MAG abundances
under the tested conditions by mapping of the obtained metagenome sequence reads to the MAGs revealed that
MAG abundance profiles were mainly shaped by the temperature but also by the OLR. However, the OLR effect
was more pronounced for the mesophilic systems as compared to the thermophilic ones. In contrast,
metatranscriptome mapping to MAGs subsequently normalized to MAG abundances showed that under
thermophilic conditions, MAGs respond to increased OLRs by shifting their transcriptional activities mainly without
adjusting their proliferation rates. This is a clear difference compared to the behavior of the microbiome under
mesophilic conditions. Here, the response to increased OLRs involved adjusting of proliferation rates and
corresponding transcriptional activities. The analysis led to the identification of MAGs positively responding to
increased OLRs. The most outstanding MAGs in this regard, obviously well adapted to higher OLRs and/or
associated conditions, were assigned to the order Clostridiales (Acetivibrio sp.) for the mesophilic biofilm and the
orders Bacteroidales (Prevotella sp. and an unknown species), Lachnospirales (Herbinix sp. and Kineothrix sp.) and
Clostridiales (Clostridium sp.) for the thermophilic biofilm. Genome-based metabolic reconstruction and
transcriptional profiling revealed that positively responding MAGs mainly are involved in hydrolysis of grass silage,
acidogenesis and / or acetogenesis.
Conclusions: An integrated -omics approach enabled the identification of new AD biofilm keystone species
featuring outstanding performance under stress conditions such as increased OLRs. Genome-based knowledge on
the metabolic potential and transcriptional activity of responsive microbiome members will contribute to the
development of improved microbiological AD management strategies for biomethanation of renewable biomass.
Keywords: Metagenome assembled genomes, Integrated -omics, Polyomics, Anaerobic digestion, Biogas,
Bioconversion, Microbial community structure, Methane, Metabolic activity
Environmental Microbiome 15, 7 (2020). doi:10.1186/s40793-020-00354-x.
10.1186/s40793-020-00354-x
http://hdl.handle.net/10033/622632
25246372
Environmental Microbiomes
2-s2.0-85082728024
SCOPUS_ID:85082728024
Anaerobic digestion
Bioconversion
Biogas
Integrated-omics
Metabolic activity
Metagenome assembled genomes
Methane
Microbial community structure
Polyomics
Impact of process temperature and organic loading rate on cellulolytic/hydrolytic biofilm microbiomes during biomethanation of ryegrass silage revealed by genome-centered metagenomics and metatranscriptomics
oai:repository.helmholtz-hzi.de:10033/6226472020-12-12T10:31:11Zcom_10033_311624com_10033_6839com_10033_620857col_10033_311625col_10033_620858
00925njm 22002777a 4500
dc
Krome, Anna K
author
Becker, Tim
author
Kehraus, Stefan
author
Schiefer, Andrea
author
Steinebach, Christian
author
Aden, Tilman
author
Frohberger, Stefan J
author
López Mármol, Álvaro
author
Kapote, Dnyaneshwar
author
Jansen, Rolf
author
Chaverra-Muñoz, Lillibeth
author
Hübner, Marc P
author
Pfarr, Kenneth
author
Hesterkamp, Thomas
author
Stadler, Marc
author
Gütschow, Michael
author
König, Gabriele M
author
Hoerauf, Achim
author
Wagner, Karl G
author
2020-11-18
Novel-antibiotics are urgently needed to combat an increase in morbidity and mortality due to resistant bacteria. The preclinical candidate corallopyronin A (CorA) is a potent antibiotic against Gram-positive and some Gram-negative pathogens for which a solid oral formulation was needed for further preclinical testing of the active pharmaceutical ingredient (API). The neat API CorA is poorly water-soluble and instable at room temperature, both crucial characteristics to be addressed and overcome for use as an oral antibiotic. Therefore, amorphous solid dispersion (ASD) was chosen as formulation principle. The formulations were prepared by spray-drying, comprising the water-soluble polymers povidone and copovidone. Stability (high-performance liquid chromatography, Fourier-transform-infrared spectroscopy, differential scanning calorimetry), dissolution (biphasic dissolution), and solubility (biphasic dissolution, Pion's T3 apparatus) properties were analyzed. Pharmacokinetic evaluations after intravenous and oral administration were conducted in BALB/c mice. The results demonstrated that the ASD formulation principle is a suitable stability- and solubility-enhancing oral formulation strategy for the API CorA to be used in preclinical and clinical trials and as a potential market product.
Pharmaceutics. 2020 Nov 18;12(11):1105. doi: 10.3390/pharmaceutics12111105.
1999-4923
33217948
10.3390/pharmaceutics12111105
http://hdl.handle.net/10033/622647
Pharmaceutics
anthelmintic
antibiotic
biphasic dissolution (BiPHa+)
copovidone (PVP/VA)
corallopyronin A (CorA)
pharmacokinetic analysis
povidone (PVP)
solubility enhanced formulation
spray-dried amorphous solid dispersion (ASD)
stability enhanced formulation
Solubility and Stability Enhanced Oral Formulations for the Anti-Infective Corallopyronin A.
oai:repository.helmholtz-hzi.de:10033/6226502020-12-15T01:34:49Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Raijmakers, Ruud P H
author
Roerink, Megan E
author
Jansen, Anne F M
author
Keijmel, Stephan P
author
Gacesa, Ranko
author
Li, Yang
author
Joosten, Leo A B
author
van der Meer, Jos W M
author
Netea, Mihai G
author
Bleeker-Rovers, Chantal P
author
Xu, Cheng-Jian
author
2020-11-26
Inflammatory markers, including 4E-BP1 (P = 9.60-16 and 1.41-7) and MMP-1 (P = 7.09-9 and 3.51-9), are significantly more expressed in both QFS and CFS patients compared to HC. Blood metabolite profiles show significant differences when comparing QFS (319 metabolites) and CFS (441 metabolites) patients to HC, and are significantly enriched in pathways like sphingolipid (P = 0.0256 and 0.0033) metabolism. When comparing QFS to CFS patients, almost no significant differences in metabolome were found. Comparison of microbiome taxonomy of QFS and CFS patients with that of HC, shows both in- and decreases in abundancies in Bacteroidetes (with emphasis on Bacteroides and Alistiples spp.), and Firmicutes and Actinobacteria (with emphasis on Ruminococcus and Bifidobacterium spp.). When we compare QFS patients to CFS patients, there is a striking resemblance and hardly any significant differences in microbiome taxonomy are found.
J Transl Med. 2020 Nov 26;18(1):448. doi: 10.1186/s12967-020-02585-5.
33243243
10.1186/s12967-020-02585-5
http://hdl.handle.net/10033/622650
1479-5876
Journal of translational medicine
CFS
Fatigue
Inflammation
Metabolome
Microbiome
Omics
Q fever
QFS
Multi-omics examination of Q fever fatigue syndrome identifies similarities with chronic fatigue syndrome.
oai:repository.helmholtz-hzi.de:10033/6226512020-12-15T01:34:59Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Vehmeijer, Florianne O L
author
Küpers, Leanne K
author
Sharp, Gemma C
author
Salas, Lucas A
author
Lent, Samantha
author
Jima, Dereje D
author
Tindula, Gwen
author
Reese, Sarah
author
Qi, Cancan
author
Gruzieva, Olena
author
Page, Christian
author
Rezwan, Faisal I
author
Melton, Philip E
author
Nohr, Ellen
author
Escaramís, Geòrgia
author
Rzehak, Peter
author
Heiskala, Anni
author
Gong, Tong
author
Tuominen, Samuli T
author
Gao, Lu
author
Ross, Jason P
author
Starling, Anne P
author
Holloway, John W
author
Yousefi, Paul
author
Aasvang, Gunn Marit
author
Beilin, Lawrence J
author
Bergström, Anna
author
Binder, Elisabeth
author
Chatzi, Leda
author
Corpeleijn, Eva
author
Czamara, Darina
author
Eskenazi, Brenda
author
Ewart, Susan
author
Ferre, Natalia
author
Grote, Veit
author
Gruszfeld, Dariusz
author
Håberg, Siri E
author
Hoyo, Cathrine
author
Huen, Karen
author
Karlsson, Robert
author
Kull, Inger
author
Langhendries, Jean-Paul
author
Lepeule, Johanna
author
Magnus, Maria C
author
Maguire, Rachel L
author
Molloy, Peter L
author
Monnereau, Claire
author
Mori, Trevor A
author
Oken, Emily
author
Räikkönen, Katri
author
Rifas-Shiman, Sheryl
author
Ruiz-Arenas, Carlos
author
Sebert, Sylvain
author
Ullemar, Vilhelmina
author
Verduci, Elvira
author
Vonk, Judith M
author
Xu, Cheng-Jian
author
Yang, Ivana V
author
Zhang, Hongmei
author
Zhang, Weiming
author
Karmaus, Wilfried
author
Dabelea, Dana
author
Muhlhausler, Beverly S
author
Breton, Carrie V
author
Lahti, Jari
author
Almqvist, Catarina
author
Jarvelin, Marjo-Riitta
author
Koletzko, Berthold
author
Vrijheid, Martine
author
Sørensen, Thorkild I A
author
Huang, Rae-Chi
author
Arshad, Syed Hasan
author
Nystad, Wenche
author
Melén, Erik
author
Koppelman, Gerard H
author
London, Stephanie J
author
Holland, Nina
author
Bustamante, Mariona
author
Murphy, Susan K
author
Hivert, Marie-France
author
Baccarelli, Andrea
author
Relton, Caroline L
author
Snieder, Harold
author
Jaddoe, Vincent W V
author
Felix, Janine F
author
2020-11-25
DNA methylation at three CpGs (cg05937453, cg25212453, and cg10040131), each in a different age range, was associated with BMI at Bonferroni significance, P < 1.06 × 10-7, with a 0.96 standard deviation score (SDS) (standard error (SE) 0.17), 0.32 SDS (SE 0.06), and 0.32 BMI SDS (SE 0.06) higher BMI per 10% increase in methylation, respectively. DNA methylation at nine additional CpGs in the cross-sectional childhood model was associated with BMI at false discovery rate significance. The strength of the associations of DNA methylation at the 187 CpGs previously identified to be associated with adult BMI, increased with advancing age across childhood and adolescence in our analyses. In addition, correlation coefficients between effect estimates for those CpGs in adults and in children and adolescents also increased. Among the top findings for each age range, we observed increasing enrichment for the CpGs that were previously identified in adults (birth Penrichment = 1; childhood Penrichment = 2.00 × 10-4; adolescence Penrichment = 2.10 × 10-7).
Genome Med. 2020 Nov 25;12(1):105. doi: 10.1186/s13073-020-00810-w.
33239103
10.1186/s13073-020-00810-w
http://hdl.handle.net/10033/622651
1756-994X
Genome medicine
Body mass index
Childhood obesity
DNA methylation
Epigenetics
DNA methylation and body mass index from birth to adolescence: meta-analyses of epigenome-wide association studies.
oai:repository.helmholtz-hzi.de:10033/6226642021-01-07T01:53:47Zcom_10033_311624com_10033_6839com_10033_620857com_10033_620618col_10033_311625col_10033_620858col_10033_620619
00925njm 22002777a 4500
dc
Schiefer, Andrea
author
Hübner, Marc P
author
Krome, Anna
author
Lämmer, Christine
author
Ehrens, Alexandra
author
Aden, Tilman
author
Koschel, Marianne
author
Neufeld, Helene
author
Chaverra-Muñoz, Lillibeth
author
Jansen, Rolf
author
Kehraus, Stefan
author
König, Gabriele M
author
Pogorevc, Domen
author
Müller, Rolf
author
Stadler, Marc
author
Hüttel, Stephan
author
Hesterkamp, Thomas
author
Wagner, Karl
author
Pfarr, Kenneth
author
Hoerauf, Achim
author
2020-12-07
Current efforts to eliminate the neglected tropical diseases onchocerciasis and lymphatic filariasis, caused by the filarial nematodes Onchocerca volvulus and Wuchereria bancrofti or Brugia spp., respectively, are hampered by lack of a short-course macrofilaricidal-adult-worm killing-treatment. Anti-wolbachial antibiotics, e.g. doxycycline, target the essential Wolbachia endosymbionts of filariae and are a safe prototype adult-worm-sterilizing and macrofilaricidal regimen, in contrast to standard treatments with ivermectin or diethylcarbamazine, which mainly target the microfilariae. However, treatment regimens of 4-5 weeks necessary for doxycycline and contraindications limit its use. Therefore, we tested the preclinical anti-Wolbachia drug candidate Corallopyronin A (CorA) for in vivo efficacy during initial and chronic filarial infections in the Litomosoides sigmodontis rodent model. CorA treatment for 14 days beginning immediately after infection cleared >90% of Wolbachia endosymbionts from filariae and prevented development into adult worms. CorA treatment of patently infected microfilaremic gerbils for 14 days with 30 mg/kg twice a day (BID) achieved a sustained reduction of >99% of Wolbachia endosymbionts from adult filariae and microfilariae, followed by complete inhibition of filarial embryogenesis resulting in clearance of microfilariae. Combined treatment of CorA and albendazole, a drug currently co-administered during mass drug administrations and previously shown to enhance efficacy of anti-Wolbachia drugs, achieved microfilarial clearance after 7 days of treatment at a lower BID dose of 10 mg/kg CorA, a Human Equivalent Dose of 1.4 mg/kg. Importantly, this combination led to a significant reduction in the adult worm burden, which has not yet been published with other anti-Wolbachia candidates tested in this model. In summary, CorA is a preclinical candidate for filariasis, which significantly reduces treatment times required to achieve sustained Wolbachia depletion, clearance of microfilariae, and inhibition of embryogenesis. In combination with albendazole, CorA is robustly macrofilaricidal after 7 days of treatment and fulfills the Target Product Profile for a macrofilaricidal drug.
PLoS Negl Trop Dis. 2020 Dec 7;14(12):e0008930. doi: 10.1371/journal.pntd.0008930.
33284808
10.1371/journal.pntd.0008930
http://hdl.handle.net/10033/622664
1935-2735
PLoS neglected tropical diseases
Corallopyronin A for short-course anti-wolbachial, macrofilaricidal treatment of filarial infections.
oai:repository.helmholtz-hzi.de:10033/6226652021-01-08T01:33:53Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Zayed, Ashraf R
author
Butmeh, Suha
author
Pecellin, Marina
author
Salah, Alaa
author
Alalam, Hanna
author
Steinert, Michael
author
Höfle, Manfred G
author
Bitar, Dina M
author
Brettar, Ingrid
author
2020-12-01
The West Bank can be considered as a high-risk area for Legionella prevalence in drinking water due to high ambient temperature, intermittent water supply, frequent pressure loss, and storage of drinking water in roof containers. To assess occurrence of Legionella species, especially L. pneumophila, in the drinking water of the West Bank, the drinking water distribution systems of eight hospitals were sampled over a period of 2.3 years covering the seasonal cycle and the major geographic regions. To gain insight into potential environmental drivers, a set of physico-chemical and microbiological parameters was recorded. Sampling included drinking water and biofilm analyzed by culture and PCR-based methods. Cultivation led to the isolation of 180 strains of L. pneumophila that were genotyped by Multi-Locus Variable Number of Tandem Repeat Analysis (MLVA). Surprisingly, the abundance of culturable L. pneumophila was low in drinking water of the sampling sites, with only three out of eight sites where Legionella was observed at all (range: 30-500 CFU/liter). By contrast, biofilm and PCR-based analyses showed a higher prevalence. Statistical analyses with physico-chemical parameters revealed a decrease of L. pneumophila abundance for water and biofilm with increasing magnesium concentrations (>30 mg/l). MLVA-genotype analysis of the L. pneumophila isolates and their spatial distribution indicated three niches characterized by distinct physico-chemical parameters and inhabited by specific consortia of genotypes. This study provides novel insights into mechanisms shaping L. pneumophila populations and triggering their abundance leading to an understanding of their genotype-specific niches and ecology in support of improved prevention measures.
Pathogens. 2020 Dec 1;9(12):1012. doi: 10.3390/pathogens9121012.
2076-0817
33271905
10.3390/pathogens9121012
http://hdl.handle.net/10033/622665
Pathogens (Basel, Switzerland)
MLVA-genotypes
ecotype
environmental factors
groundwater
magnesium
niche
Biogeography and Environmental Drivers of Abundance and Genotype Composition Across the West Bank: Relevance of a Genotype-Based Ecology for Understanding Occurrence.
oai:repository.helmholtz-hzi.de:10033/6226862021-01-16T01:44:15Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Kaipa, Jagan Mohan
author
Starkuviene, Vytaute
author
Erfle, Holger
author
Eils, Roland
author
Gladilin, Evgeny
author
2020-12-16
Silibinin (SIL), a natural flavonolignan from the milk thistle (Silybum marianum), is known to exhibit remarkable hepatoprotective, antineoplastic and EMT inhibiting effects in different cancer cells by targeting multiple molecular targets and pathways. However, the predominant majority of previous studies investigated effects of this phytocompound in a one particular cell line. Here, we carry out a systematic analysis of dose-dependent viability response to SIL in five non-small cell lung cancer (NSCLC) lines that gradually differ with respect to their intrinsic EMT stage. By correlating gene expression profiles of NSCLC cell lines with the pattern of their SIL IC50 response, a group of cell cycle, survival and stress responsive genes, including some prominent targets of STAT3 (BIRC5, FOXM1, BRCA1), was identified. The relevancy of these computationally selected genes to SIL viability response of NSCLC cells was confirmed by the transient knockdown test. In contrast to other EMT-inhibiting compounds, no correlation between the SIL IC50 and the intrinsic EMT stage of NSCLC cells was observed. Our experimental results show that SIL viability response of differently constituted NSCLC cells is linked to a subnetwork of tightly interconnected genes whose transcriptomic pattern can be used as a benchmark for assessment of individual SIL sensitivity instead of the conventional EMT signature. Insights gained in this study pave the way for optimization of customized adjuvant therapy of malignancies using Silibinin.
PeerJ. 2020 Dec 16;8:e10373. doi: 10.7717/peerj.10373.
2167-8359
33362957
10.7717/peerj.10373
http://hdl.handle.net/10033/622686
PeerJ
Adjuvant cancer therapy
Custom drug response
Drug susceptibility network
Silibinin
Transcriptome profiling
Transcriptome profiling reveals Silibinin dose-dependent response network in non-small lung cancer cells.
oai:repository.helmholtz-hzi.de:10033/6227042021-01-28T01:39:24Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Li, Zhaopeng
author
Rinas, Ursula
author
2020-04-06
Background: Recombinant protein production can be stressful to the host organism. The extent of stress is determined by the specific properties of the recombinant transcript and protein, by the rates of transcription and translation, and by the environmental conditions encountered during the production process.
Results: The impact of the transcription of the T7-promoter controlled genes encoding human basic fibroblast growth factor (hFGF-2) and green fluorescent protein (GFP) as well as the translation into the recombinant protein on the growth properties of the production host E. coli BL21(DE3) were investigated. This was done by using expression vectors where the promoter region or the ribosome binding site(s) or both were removed. It is shown that already transcription without protein translation imposes a metabolic burden on the host cell. Translation of the transcript into large amounts of a properly folded protein does not show any effect on cell growth in the best case, e.g. high-level production of GFP in Luria-Bertani medium. However, translation appears to contribute to the metabolic burden if it is connected to protein folding associated problems, e.g. inclusion body formation.
Conclusion: The so-called metabolic burden of recombinant protein production is mainly attributed to transcription but can be enhanced through translation and those processes following translation (e.g. protein folding and degradation, heat-shock responses).
Microb Cell Fact. 2020 Apr 6;19(1):83. doi: 10.1186/s12934-020-01343-y.
32252765
10.1186/s12934-020-01343-y
http://hdl.handle.net/10033/622704
1475-2859
Microbial cell factories
Escherichia coli
Metabolic burden
Recombinant protein production
Transcriptional burden
Recombinant protein production associated growth inhibition results mainly from transcription and not from translation.
oai:repository.helmholtz-hzi.de:10033/6227132021-02-03T09:20:34Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Abdissa, Ketema
author
Ruangkiattikul, Nanthapon
author
Ahrend, Wiebke
author
Nerlich, Andreas
author
Beineke, Andreas
author
Laarmann, Kristin
author
Janze, Nina
author
Lobermeyer, Ulrike
author
Suwandi, Abdulhadi
author
Falk, Christine
author
Schleicher, Ulrike
author
Weiss, Siegfried
author
Bogdan, Christian
author
Goethe, Ralph
author
2020-05-14
Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne's disease (JD), an incurable chronic intestinal bowel disease in ruminants. JD occurs worldwide and causes enormous economic burden in dairy industry. Research on JD pathobiology is hampered by its complexity which cannot completely be mimicked by small animal models. As a model the mouse allows dissecting some pathogenicity features of MAP. However, for unknown reasons MAP exhibits reduced growth in granulomas of infected mice compared to other Mycobacterium avium subspecies. Here, we characterized immune reactions of MAP-infected C57BL/6 mice. After infection, mice appeared fully immunocompetent. A strong antigen-specific T cell response was elicited indicated by IFNγ production of splenic T cells re-stimulated with MAP antigens. Function of splenic dendritic cells and proliferation of adoptively transferred antigen-specific CD4+ T cells was unaltered. Isolated splenic myeloid cells from infected mice revealed that MAP resides in CD11b+ macrophages. Importantly, sorted CD11b+CD11c- cells expressed high level of type 2 nitric oxide synthase (NOS2) but only low levels of pro- and anti-inflammatory cytokines. Correspondingly, MAP-infected MAC2 expressing myeloid cells in spleen and liver granuloma displayed strong expression of NOS2. In livers of infected Nos2-/-mice higher bacterial loads, more granuloma and larger areas of tissue damage were observed 5 weeks post infection compared to wild type mice. In vitro, MAP was sensitive to NO released by a NO-donor. Thus, a strong T cell response and concomitant NOS2/NO activity appears to control MAP infection, but allows development of chronicity and pathogen persistence. A similar mechanism might explain persistence of MAP in ruminants.
Virulence. 2020 Dec;11(1):465-481. doi: 10.1080/21505594.2020.1763055.
32408806
10.1080/21505594.2020.1763055
http://hdl.handle.net/10033/622713
2150-5608
Virulence
Johne’s disease
Mycobacterium
inducible or type 2 nitric oxide synthase
macrophage
paratuberculosis
Relevance of inducible nitric oxide synthase for immune control of Mycobacterium avium subspecies paratuberculosis infection in mice.
oai:repository.helmholtz-hzi.de:10033/6227422021-02-16T03:17:51Zcom_10033_311624com_10033_6839com_10033_311308com_10033_338554col_10033_311625col_10033_620721col_10033_559591col_10033_338544
00925njm 22002777a 4500
dc
Chaoprasid, Paweena
author
Lukat, Peer
author
Mühlen, Sabrina
author
Heidler, Thomas
author
Gazdag, Emerich-Mihai
author
Dong, Shuangshuang
author
Bi, Wenjie
author
Rüter, Christian
author
Kirchenwitz, Marco
author
Steffen, Anika
author
Jänsch, Lothar
author
Stradal, Theresia E B
author
Dersch, Petra
author
Blankenfeldt, Wulf
author
2021-01-07
Cytotoxic necrotizing factors (CNFs) are bacterial single-chain exotoxins that modulate cytokinetic/oncogenic and inflammatory processes through activation of host cell Rho GTPases. To achieve this, they are secreted, bind surface receptors to induce endocytosis and translocate a catalytic unit into the cytosol to intoxicate host cells. A three-dimensional structure that provides insight into the underlying mechanisms is still lacking. Here, we determined the crystal structure of full-length Yersinia pseudotuberculosis CNFY . CNFY consists of five domains (D1-D5), and by integrating structural and functional data, we demonstrate that D1-3 act as export and translocation module for the catalytic unit (D4-5) and for a fused β-lactamase reporter protein. We further found that D4, which possesses structural similarity to ADP-ribosyl transferases, but had no equivalent catalytic activity, changed its position to interact extensively with D5 in the crystal structure of the free D4-5 fragment. This liberates D5 from a semi-blocked conformation in full-length CNFY , leading to higher deamidation activity. Finally, we identify CNF translocation modules in several uncharacterized fusion proteins, which suggests their usability as a broad-specificity protein delivery tool.
EMBO J. 2021 Jan 7:e105202. doi: 10.15252/embj.2020105202. Epub ahead of print.
33410511
10.15252/embj.2020105202
http://hdl.handle.net/10033/622742
1460-2075
The EMBO journal
Yersinia
AB-toxin
ADP-ribosyl transferase
CNF
DUF4765
Crystal structure of bacterial cytotoxic necrotizing factor CNFy reveals molecular building blocks for intoxication.
oai:repository.helmholtz-hzi.de:10033/6227592021-03-02T01:32:20Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Fernández, Nathalie
author
2020-05-11
No abstract available
Int J Infect Dis. 2020 Jul;96:630. doi: 10.1016/j.ijid.2020.04.084. Epub 2020 May 11.
32437940
10.1016/j.ijid.2020.04.084
http://hdl.handle.net/10033/622759
1878-3511
International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases
Misinterpretation of the odds ratios.
oai:repository.helmholtz-hzi.de:10033/6228662021-05-12T02:38:58Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Penkert, Judith
author
Märtens, Andre
author
Seifert, Martin
author
Auber, Bernd
author
Derlin, Katja
author
Hille-Betz, Ursula
author
Hörmann, Philipp
author
Klopp, Norman
author
Prokein, Jana
author
Schlicker, Lisa
author
Wacker, Frank
author
Wallaschek, Hannah
author
Schlegelberger, Brigitte
author
Hiller, Karsten
author
Ripperger, Tim
author
Illig, Thomas
author
2021-04-07
Individuals carrying a pathogenic germline variant in the breast cancer predisposition gene BRCA1 (gBRCA1+) are prone to developing breast cancer. Apart from its well-known role in DNA repair, BRCA1 has been shown to powerfully impact cellular metabolism. While, in general, metabolic reprogramming was named a hallmark of cancer, disrupted metabolism has also been suggested to drive cancer cell evolution and malignant transformation by critically altering microenvironmental tissue integrity. Systemic metabolic effects induced by germline variants in cancer predisposition genes have been demonstrated before. Whether or not systemic metabolic alterations exist in gBRCA1+ individuals independent of cancer incidence has not been investigated yet. We therefore profiled the plasma metabolome of 72 gBRCA1+ women and 72 age-matched female controls, none of whom (carriers and non-carriers) had a prior cancer diagnosis and all of whom were cancer-free during the follow-up period. We detected one single metabolite, pyruvate, and two metabolite ratios involving pyruvate, lactate, and a metabolite of yet unknown structure, significantly altered between the two cohorts. A machine learning signature of metabolite ratios was able to correctly distinguish between gBRCA1+ and controls in ~82%. The results of this study point to innate systemic metabolic differences in gBRCA1+ women independent of cancer incidence and raise the question as to whether or not constitutional alterations in energy metabolism may be involved in the etiology of BRCA1-associated breast cancer.
Front Oncol. 2021 Apr 7;11:627217. doi: 10.3389/fonc.2021.627217.
2234-943X
33898308
10.3389/fonc.2021.627217
http://hdl.handle.net/10033/622866
Frontiers in oncology
BRCA1 germline mutation
HIF1 alpha
NAD+ balance
aerobic glycolysis
breast cancer
energy metabolism
lactate
plasma metabolome
Plasma Metabolome Signature Indicative of Germline Status Independent of Cancer Incidence.
oai:repository.helmholtz-hzi.de:10033/6228782021-05-19T04:38:49Zcom_10033_311624com_10033_6839col_10033_311625
00925njm 22002777a 4500
dc
Zayed, Ashraf R
author
Pecellin, Marina
author
Jaber, Lina
author
Butmeh, Suha
author
Bahader, Shereen A
author
Steinert, Michael
author
Höfle, Manfred G
author
Brettar, Ingrid
author
Bitar, Dina M
author
2021-04-01
Legionella pneumophila is the causative agent of Legionnaires' disease. Due to the hot climate and intermittent water supply, the West Bank, Palestine, can be considered a high-risk area for this often fatal atypical pneumonia. L. pneumophila occurs in biofilms of natural and man-made freshwater environments, where it infects and replicates intracellularly within protozoa. To correlate the genetic diversity of the bacteria in the environment with their virulence properties for protozoan and mammalian host cells, 60 genotyped isolates from hospital water systems in the West Bank were analyzed. The L. pneumophila isolates were previously genotyped by high resolution Multi Locus Variable Number of Tandem Repeat Analysis (MLVA-8(12)) and sorted according to their relationship in clonal complexes (VACC). Strains of relevant genotypes and VACCs were compared according to their capacity to infect Acanthamoeba castellanii and THP-1 macrophages, and to mediate pore-forming cytotoxicity in sheep red blood cells (sRBCs). Based on a previous detailed analysis of the biogeographic distribution and abundance of the MLVA-8(12)-genotypes, the focus of the study was on the most abundant L. pneumophila- genotypes Gt4(17), Gt6 (18) and Gt10(93) and the four relevant clonal complexes [VACC1, VACC2, VACC5 and VACC11]. The highly abundant genotypes Gt4(17) and Gt6(18) are affiliated with VACC1 and sequence type (ST)1 (comprising L. pneumophila str. Paris), and displayed seroroup (Sg)1. Isolates of these two genotypes exhibited significantly higher virulence potentials compared to other genotypes and clonal complexes in the West Bank. Endemic for the West Bank was the clonal complex VACC11 (affiliated with ST461) represented by three relevant genotypes that all displayed Sg6. These genotypes unique for the West Bank showed a lower infectivity and cytotoxicity compared to all other clonal complexes and their affiliated genotypes. Interestingly, the L. pneumophila serotypes ST1 and ST461 were previously identified by in situ-sequence based typing (SBT) as main causative agents of Legionnaires' disease (LD) in the West Bank at a comparable level. Overall, this study demonstrates the site-specific regional diversity of L. pneumophila genotypes in the West Bank and suggests that a combination of MLVA, cellular infection assays and hierarchical agglomerative cluster analysis allows an improved genotype-based risk assessment.
Pathogens. 2021 Apr 1;10(4):417. doi: 10.3390/pathogens10040417.
2076-0817
33915921
10.3390/pathogens10040417
http://hdl.handle.net/10033/622878
Pathogens (Basel, Switzerland)
Gt10(93)
Legionella pneumophila
MLVA
VACC11
clonal complex
genotype
virulence
Cytotoxicity, Intracellular Replication, and Contact-Dependent Pore Formation of Genotyped Environmental Isolates from Hospital Water Systems in the West Bank, Palestine.
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