2024-03-28T09:31:11Zhttp://repository.helmholtz-hzi.de/oai/requestoai:repository.helmholtz-hzi.de:10033/6213362020-06-30T13:26:39Zcom_10033_196529com_10033_56876com_10033_6839col_10033_196534col_10033_621495
Homologous recombination mediates stable Fah gene integration and phenotypic correction in tyrosinaemia mouse-model.
Junge, Norman
Yuan, Qinggong
Vu, Thu Huong
Krooss, Simon
Bednarski, Christien
Balakrishnan, Asha
Cathomen, Toni
Manns, Michael P
Baumann, Ulrich
Sharma, Amar Deep
Ott, Michael
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
To stably correct tyrosinaemia in proliferating livers of fumarylacetoacetate-hydrolase knockout (Fah-/-)mice by homologous-recombination-mediated targeted addition of theFahgene.
2018-04-05
2018-04-05
2018-02-27
Article
Homologous recombination mediates stable Fah gene integration and phenotypic correction in tyrosinaemia mouse-model. 2018, 10 (2):277-286 World J Hepatol
1948-5182
29527263
10.4254/wjh.v10.i2.277
http://hdl.handle.net/10033/621336
World journal of hepatology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6214962019-08-30T11:30:31Zcom_10033_6839col_10033_621495
Targeting Antigens to Dendritic Cells the DC-Specific-ICAM3-Grabbing-Nonintegrin Receptor Induces Strong T-Helper 1 Immune Responses.
Velasquez, Lis Noelia
Stüve, Philipp
Gentilini, Maria Virginia
Swallow, Maxine
Bartel, Judith
Lycke, Nils Yngve
Barkan, Daniel
Martina, Mariana
Lujan, Hugo D
Kalay, Hakan
van Kooyk, Yvette
Sparwasser, Tim D
Berod, Luciana
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
Ag85B
DC-specific-ICAM3-grabbing-nonintegrin
dendritic cells
tuberculosis
vaccine
Tuberculosis remains a major global health problem and efforts to develop a more effective vaccine have been unsuccessful so far. Targeting antigens (Ags) to dendritic cells
(DCs) in vivo has emerged as a new promising vaccine strategy. In this approach, Ags
are delivered directly to DCs via antibodies that bind to endocytic cell-surface receptors.
Here, we explored DC-specifc-ICAM3-grabbing-nonintegrin (DC-SIGN) targeting as
a potential vaccine against tuberculosis. For this, we made use of the hSIGN mouse
model that expresses human DC-SIGN under the control of the murine CD11c promoter.
We show that in vitro and in vivo delivery of anti-DC-SIGN antibodies conjugated to
Ag85B and peptide 25 of Ag85B in combination with anti-CD40, the fungal cell wall
component zymosan, and the cholera toxin-derived fusion protein CTA1-DD induces
strong Ag-specifc CD4+ T-cell responses. Improved anti-mycobacterial immunity was
accompanied by increased frequencies of Ag-specifc IFN-γ+ IL-2+ TNF-α+ polyfunctional
CD4+ T cells in vaccinated mice compared with controls. Taken together, in this study
we provide the proof of concept that the human DC-SIGN receptor can be effciently
exploited for vaccine purposes to promote immunity against mycobacterial infections.
2018-09-25
2018-09-25
2018-01-01
Article
1664-3224
29662482
10.3389/fimmu.2018.00471
http://hdl.handle.net/10033/621496
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Attribution-NonCommercial-ShareAlike 3.0 United States
oai:repository.helmholtz-hzi.de:10033/6215172019-08-30T11:29:46Zcom_10033_6839col_10033_621495
Sialylation Is Dispensable for Early Murine Embryonic Development in Vitro.
Abeln, Markus
Borst, Kristina M
Cajic, Samanta
Thiesler, Hauke
Kats, Elina
Albers, Iris
Kuhn, Maike
Kaever, Volkhard
Rapp, Erdmann
Münster-Kühnel, Anja
Weinhold, Birgit
TWINCORE, Zentrum für experimentelle und klinischeInfektionsforschung GmbH, Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
CMP-sialic acid synthase
differentiation
glycosylation
metabolism
sialic acids
The negatively charged nonulose sialic acid (Sia) is essential for murine development in vivo. In order to elucidate the impact of sialylation on differentiation processes in the absence of maternal influences, we generated mouse embryonic stem cell (mESC) lines that lack CMP-Sia synthetase (CMAS) and thereby the ability to activate Sia to CMP-Sia. Loss of CMAS activity resulted in an asialo cell surface accompanied by an increase in glycoconjugates with terminal galactosyl and oligo-LacNAc residues, as well as intracellular accumulation of free Sia. Remarkably, these changes did not impact intracellular metabolites or the morphology and transcriptome of pluripotent mESC lines. Moreover, the capacity of Cmas
2018-10-12
2018-10-12
2017-07-04
Article
1439-7633
28374933
10.1002/cbic.201700083
http://hdl.handle.net/10033/621517
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Attribution-NonCommercial-ShareAlike 3.0 United States
oai:repository.helmholtz-hzi.de:10033/6215832019-08-30T11:28:18Zcom_10033_6839col_10033_621495
C-Type Lectin Receptor (CLR)-Fc Fusion Proteins As Tools to Screen for Novel CLR/Bacteria Interactions: An Exemplary Study on Preselected Isolates.
Mayer, Sabine
Moeller, Rebecca
Monteiro, João T
Ellrott, Kerstin
Josenhans, Christine
Lepenies, Bernd
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
C-type lectin receptors
Campylobacter jejuni
Dectin-1 receptor
ELISA assay
confocal microscopy
flow cytometry
innate immunity
screening tools
C-type lectin receptors (CLRs) are carbohydrate-binding receptors that recognize
their ligands often in a Ca2+-dependent manner. Upon ligand binding, myeloid CLRs
in innate immunity trigger or inhibit a variety of signaling pathways, thus initiating or
modulating effector functions such as cytokine production, phagocytosis, and antigen
presentation. CLRs bind to various pathogens, including viruses, fungi, parasites, and
bacteria. The bacterium Campylobacter jejuni (C. jejuni) is a very frequent Gram-negative
zoonotic pathogen of humans, causing severe intestinal symptoms. Interestingly,
C. jejuni expresses several glycosylated surface structures, for example, the capsular
polysaccharide (CPS), lipooligosaccharide (LOS), and envelope proteins. This “Methods”
paper describes applications of CLR–Fc fusion proteins to screen for yet unknown
CLR/bacteria interactions using C. jejuni as an example. ELISA-based detection of
CLR/bacteria interactions allows a frst prescreening that is further confrmed by flow
cytometry-based binding analysis and visualized using confocal microscopy. By
applying these methods, we identifed Dectin-1 as a novel CLR recognizing two selected
C. jejuni isolates with different LOS and CPS genotypes. In conclusion, the heredescribed applications of CLR–Fc fusion proteins represent useful methods to screen for
and identify novel CLR/bacteria interactions.
2018-11-26
2018-11-26
2018-01-01
Article
1664-3224
29487596
10.3389/fimmu.2018.00213
http://hdl.handle.net/10033/621583
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Attribution-NonCommercial-ShareAlike 3.0 United States
oai:repository.helmholtz-hzi.de:10033/6216322019-08-30T11:31:46Zcom_10033_6839col_10033_621495
Cell therapy products: focus on issues with manufacturing and quality control of chimeric antigen receptor T-cell therapies
Eyles, Jim E
Vessillier, Sandrine
Jones, Anika
Stacey, Glyn
Schneider, Christian K
Price, Jack
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
2019-01-07
2019-01-07
2018-12-17
Article
02682575
10.1002/jctb.5829
http://hdl.handle.net/10033/621632
http://doi.wiley.com/10.1002/jctb.5829
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
oai:repository.helmholtz-hzi.de:10033/6216792019-08-30T11:32:39Zcom_10033_620601com_10033_6839col_10033_620602col_10033_621495
Cell therapy products: focus on issues with manufacturing and quality control of chimeric antigen receptor T-cell therapies
Eyles, Jim E
Vessillier, Sandrine
Jones, Anika
Stacey, Glyn
Schneider, Christian K
Price, Jack
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Recent accelerated approvals of Chimeric Antigen Receptor T‐cell (CAR‐T) therapies targeting refractory haematological malignancies underscore the potential for this novel technology platform to provide new therapeutic options for oncology areas with high unmet medical needs. However, these powerful ‘living drugs’ are markedly different to conventional small molecule and biologic therapies on several levels. The highly complex nature and varied composition of CAR‐T based products still requires considerable investigation to resolve the best approaches to ensure reproducible and cost‐effective manufacture, clinical development, and application. This review will focus on key issues for manufacturing and quality control of these exciting new therapeutic modalities, preceded by a brief description of CAR principals and clinical development considerations. © 2018 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
2019-02-05
2019-02-05
Article
02682575
10.1002/jctb.5829
http://hdl.handle.net/10033/621679
Journal of Chemical Technology and Biotechnology
en
http://doi.wiley.com/10.1002/jctb.5829
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Wiley-Blackwell
oai:repository.helmholtz-hzi.de:10033/6216822019-08-30T11:32:39Zcom_10033_56876com_10033_6839col_10033_56896col_10033_621495
C-X-C Motif Chemokine Receptor 4 Blockade Promotes Tissue Repair After Myocardial Infarction by Enhancing Regulatory T Cell Mobilization and Immune-Regulatory Function.
Wang, Yong
Dembowsky, Klaus
Chevalier, Eric
Stüve, Philipp
Korf-Klingebiel, Mortimer
Lochner, Matthias
Napp, L Christian
Frank, Heike
Brinkmann, Eva
Kanwischer, Anna
Bauersachs, Johann
Gyongyosi, Mariann
Sparwasser, Tim
Wollert, Kai C
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
CXCR4 antagonist
Regulatory T cells
Acute myocardial infarction (MI) elicits an inflammatory response that drives tissue repair and adverse cardiac remodeling. Inflammatory cell trafficking after MI is controlled by C X-C motif chemokine ligand 12 (CXCL12) and its receptor, C-X-C motif chemokine receptor 4 (CXCR4). CXCR4 antagonists mobilize inflammatory cells and promote infarct repair, but the cellular mechanisms are unclear. We investigated the therapeutic potential and mode of action of the peptidic macrocycle CXCR4 antagonist POL5551 in mice with reperfused MI. We applied cell depletion and adoptive transfer strategies using lymphocyte-deficient Rag1 knockout mice; DEREG mice, which express a diphtheria toxin receptor-enhanced green fluorescent protein fusion protein under the control of the promoter/enhancer region of the regulatory T (T Intraperitoneal POL5551 injections in wild-type mice (8 mg/kg at 2, 4, 6, and 8 d) enhanced angiogenesis in the infarct border-zone, reduced scar size, and attenuated left ventricular remodeling and contractile dysfunction at 28 d. Treatment effects were absent in splenectomized wild-type mice, Rag1 knockout mice, and T Our data confirm CXCR4 blockade as a promising treatment strategy after MI. We identify dendritic cell-primed splenic T
2019-02-06
2019-02-06
2019-01-30
Article
1524-4539
30696265
10.1161/CIRCULATIONAHA.118.036053
http://hdl.handle.net/10033/621682
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Lippinscott, Williams & Wilkins; American Heart Association
oai:repository.helmholtz-hzi.de:10033/6216862019-08-30T11:33:03Zcom_10033_6839col_10033_621495
Efficient oral vaccination by bioengineering virus-like particles with protozoan surface proteins.
Serradell, Marianela C
Rupil, Lucía L
Martino, Román A
Prucca, César G
Carranza, Pedro G
Saura, Alicia
Fernández, Elmer A
Gargantini, Pablo R
Tenaglia, Albano H
Petiti, Juan P
Tonelli, Renata R
Reinoso-Vizcaino, Nicolás
Echenique, José
Berod, Luciana
Piaggio, Eliane
Bellier, Bertrand
Sparwasser, Tim
Klatzmann, David
Luján, Hugo D
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Intestinal and free-living protozoa, such as Giardia lamblia, express a dense coat of variant-specific surface proteins (VSPs) on trophozoites that protects the parasite inside the host's intestine. Here we show that VSPs not only are resistant to proteolytic digestion and extreme pH and temperatures but also stimulate host innate immune responses in a TLR-4 dependent manner. We show that these properties can be exploited to both protect and adjuvant vaccine antigens for oral administration. Chimeric Virus-like Particles (VLPs) decorated with VSPs and expressing model surface antigens, such as influenza virus hemagglutinin (HA) and neuraminidase (NA), are protected from degradation and activate antigen presenting cells in vitro. Orally administered VSP-pseudotyped VLPs, but not plain VLPs, generate robust immune responses that protect mice from influenza infection and HA-expressing tumors. This versatile vaccine platform has the attributes to meet the ultimate challenge of generating safe, stable and efficient oral vaccines.
2019-02-12
2019-02-12
2019-01-21
Article
2041-1723
30664644
10.1038/s41467-018-08265-9
http://hdl.handle.net/10033/621686
Nature Communications
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Springer-Nature
oai:repository.helmholtz-hzi.de:10033/6217682019-08-30T11:36:54Zcom_10033_6839col_10033_621495
TGFβ-activation by dendritic cells drives Th17 induction and intestinal contractility and augments the expulsion of the parasite Trichinella spiralis in mice.
Steel, Nicola
Faniyi, Aduragbemi A
Rahman, Sayema
Swietlik, Stefanie
Czajkowska, Beata I
Chan, Bethany T
Hardgrave, Alexander
Steel, Anthony
Sparwasser, Tim D
Assas, Mushref B
Grencis, Richard K
Travis, Mark A
Worthington, John J
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH, Feodor-Lynen-Str.7,30625 Hannover, Germany.
Helminths are highly prevalent metazoan parasites that infect over a billion of the world’s population. Hosts have evolved numerous mechanisms to drive the expulsion of these parasites via Th2-driven immunity, but these responses must be tightly controlled to prevent equally devastating immunopathology. However, mechanisms that regulate this balance are still unclear. Here we show that the vigorous Th2 immune response driven by the small intestinal helminth Trichinella spiralis, is associated with increased TGFβ signalling responses in CD4+ T-cells. Mechanistically, enhanced TGFβ signalling in CD4+ T-cells is dependent on dendritic cell-mediated TGFβ activation which requires expression of the integrin αvβ8. Importantly, mice lacking integrin αvβ8 on DCs had a delayed ability to expel a T. spiralis infection, indicating an important functional role for integrin αvβ8-mediated TGFβ activation in promoting parasite expulsion. In addition to maintaining regulatory T-cell responses, the CD4+ T-cell signalling of this pleiotropic cytokine induces a Th17 response which is crucial in promoting the intestinal muscle hypercontractility that drives worm expulsion. Collectively, these results provide novel insights into intestinal helminth expulsion beyond that of classical Th2 driven immunity, and highlight the importance of IL-17 in intestinal contraction which may aid therapeutics to numerous diseases of the intestine.
2019-05-10
2019-05-10
2019-01-01
Article
PLoS Pathog. 2019 Apr 18;15(4):e1007657. doi: 10.1371/journal.ppat.1007657. eCollection 2019 Apr.
1553-7374
30998782
10.1371/journal.ppat.1007657
http://hdl.handle.net/10033/621768
PLoS pathogens
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
PLOS
oai:repository.helmholtz-hzi.de:10033/6218052019-08-30T11:24:26Zcom_10033_6839col_10033_621495
TLR7 Controls VSV Replication in CD169 SCS Macrophages and Associated Viral Neuroinvasion.
Solmaz, Gülhas
Puttur, Franz
Francozo, Marcela
Lindenberg, Marc
Guderian, Melanie
Swallow, Maxine
Duhan, Vikas
Khairnar, Vishal
Kalinke, Ulrich
Ludewig, Burkhard
Clausen, Björn E
Wagner, Hermann
Lang, Karl S
Sparwasser, Tim D
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Toll-like receptor 7
conditional knock-out mice
innate immunity
pattern recognition receptors
subcapsular sinus macrophages
subcutaneous infection
vesicular stomatitis virus
virus replication
Vesicular stomatitis virus (VSV) is an insect-transmitted rhabdovirus that is neurovirulent in mice. Upon peripheral VSV infection, CD169+ subcapsular sinus (SCS) macrophages capture VSV in the lymph, support viral replication, and prevent CNS neuroinvasion. To date, the precise mechanisms controlling VSV infection in SCS macrophages remain incompletely understood. Here, we show that Toll-like receptor-7 (TLR7), the main sensing receptor for VSV, is central in controlling lymph-borne VSV infection. Following VSV skin infection, TLR7-/- mice display significantly less VSV titers in the draining lymph nodes (dLN) and viral replication is attenuated in SCS macrophages. In contrast to effects of TLR7 in impeding VSV replication in the dLN, TLR7-/- mice present elevated viral load in the brain and spinal cord highlighting their susceptibility to VSV neuroinvasion. By generating novel TLR7 floxed mice, we interrogate the impact of cell-specific TLR7 function in anti-viral immunity after VSV skin infection. Our data suggests that TLR7 signaling in SCS macrophages supports VSV replication in these cells, increasing LN infection and may account for the delayed onset of VSV-induced neurovirulence observed in TLR7-/- mice. Overall, we identify TLR7 as a novel and essential host factor that critically controls anti-viral immunity to VSV. Furthermore, the novel mouse model generated in our study will be of valuable importance to shed light on cell-intrinsic TLR7 biology in future studies.
2019-06-07
2019-06-07
2019-01-01
Article
Front Immunol. 2019 Mar 15;10:466. doi: 10.3389/fimmu.2019.00466. eCollection 2019.
1664-3224
30930901
http://hdl.handle.net/10033/621805
Frontiers in immunology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Frontiers
oai:repository.helmholtz-hzi.de:10033/6218542019-08-30T11:24:28Zcom_10033_6839col_10033_621495
A combined in silico and in vitro study on mouse Serpina1a antitrypsin-deficiency mutants.
Eggenschwiler, Reto
Patronov, Atanas
Hegermann, Jan
Fráguas-Eggenschwiler, Mariane
Wu, Guangming
Cortnumme, Leon
Ochs, Matthias
Antes, Iris
Cantz, Tobias
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Certain point-mutations in the human SERPINA1-gene can cause severe α1-antitrypsin-deficiency (A1AT-D). Affected individuals can suffer from loss-of-function lung-disease and from gain-of-function liver-disease phenotypes. However, age of onset and severity of clinical appearance is heterogeneous amongst carriers, suggesting involvement of additional genetic and environmental factors. The generation of authentic A1AT-D mouse-models has been hampered by the complexity of the mouse Serpina1-gene locus and a model with concurrent lung and liver-disease is still missing. Here, we investigate point-mutations in the mouse Serpina1a antitrypsin-orthologue, which are homolog-equivalent to ones known to cause severe A1AT-D in human. We combine in silico and in vitro methods and we find that analyzed mutations do introduce potential disease-causing properties into Serpina1a. Finally, we show that introduction of the King’s-mutation causes inactivation of neutrophil elastase inhibitory-function in both, mouse and human antitrypsin, while the mouse Z-mutant retains activity. This work paves the path to generation of better A1AT-D mouse-models.
2019-07-09
2019-07-09
2019-05-16
Article
Sci Rep. 2019 May 16;9(1):7486. doi: 10.1038/s41598-019-44043-3.
2045-2322
31097772
10.1038/s41598-019-44043-3
http://hdl.handle.net/10033/621854
Scientific reports
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Springer-Nature
oai:repository.helmholtz-hzi.de:10033/6219482019-09-19T01:31:01Zcom_10033_6839col_10033_621495
Commonly setting biological standards in rare diseases
O’Connor, Daniel J.
Buckland, Jenny
Almond, Neil
Boyle, Jennifer
Coxon, Carmen
Gaki, Eleni
Martin, Javier
Mattiuzzo, Giada
Metcalfe, Clive
Page, Mark
Rose, Nicola
Valdazo-Gonzalez, Begona
Zhao, Yuan
Schneider, Christian K.
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
diagnostics
prophylaxis
Rare diseases
reference materials
standardization
standardization institutions
surveillance
therapeutics
Introduction: Standardization is important across the life cycle of medicinal products, supporting the diagnosis, treatment, and prevention of a wide range of diseases. For rare diseases, standardization is even more important, as patient groups are small, presenting significant challenges in the design, conduct, analysis, and interpretation of clinical studies. It is here that standardization institutions, including the UK’s National Institute for Biological Standards and Control (NIBSC), can have a key role.
Areas covered: A considerable proportion of NIBSC’s work supports the better understanding, diagnosis, treatment, and prevention of rare diseases. NIBSC is also part of the UK’s Medicines and Healthcare products Regulatory Agency (MHRA), creating an agency that is uniquely placed to combine scientific and regulatory expertize for the benefit of public health. This review provides an overview of NIBSC’s work in rare diseases and highlights the positive impact of the work of standardization institutions in this field.
Expert opinion: Standardization in product development is key for patients with rare diseases. The work of standardization institutions is increasingly being recognized as crucial for supporting scientific and clinical advancements, and early and collaborative interactions can provide drug developers with the necessary expertize, when standards matter most.
2019-09-18
2019-09-18
2019-01-01
Article
10.1080/21678707.2019.1652598
https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85070942310&origin=inward
http://hdl.handle.net/10033/621948
Expert Opinion on Orphan Drugs
2-s2.0-85070942310
SCOPUS_ID:85070942310
en
Expert Opinion on Orphan Drugs
7-8
7
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Taylor& Francis
oai:repository.helmholtz-hzi.de:10033/6219882019-10-22T01:39:28Zcom_10033_6839col_10033_621495
Parasites in brains of wild rodents (Arvicolinae and Murinae) in the city of Leipzig, Germany
Waindok, Patrick
Özbakış-Beceriklisoy, Gökben
Janecek-Erfurth, Elisabeth
Springer, Andrea
Pfeffer, Martin
Leschnik, Michael
Strube, Christina
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Apodemus flavicollis
Frenkelia glareoli
Myodes glareolus
Toxocara
Toxoplasma
Zoonosis
Small rodents serve as intermediate or paratenic hosts for a variety of parasites and may participate in thetransmission of these parasites into synanthropic cycles. Parasites with neuroinvasive stages, such asToxoplasmagondiiorToxocara canis, can cause detrimental damage in the brain of intermediate or paratenic hosts.Therefore, the occurrence of neuroinvasive parasite stages was evaluated in brains of wild rodents captured inthe city of Leipzig, Germany. In addition, a few specimens from the cities of Hanover, Germany, and Vienna,Austria were included, resulting in a total of 716 rodents collected between 2011 and 2016. Brains were in-vestigated for parasitic stages by microscopic examination of native tissue, artificially digested tissue as well asGiemsa-stained digestion solution to verify positive results. Infective stages of zoonotic ascarids or other hel-minths were not detected in any sample, while coccidian cysts were found in 10.1% (95% CI: 7.9–12.5%; 72/716) of examined brains. The most abundant rodent species in the study was the bank vole (Myodes glareolus;Arvicolinae), showing an infection rate with cerebral cysts of 13.9% (95% CI: 11.0–17.8%; 62/445), while 2.7%(95% CI: 1.0–5.8%; 6/222) of yellow-necked mice (Apodemusflavicollis; Murinae) were infected. Generalizedlinear modelling revealed a statistically significant difference in prevalence betweenM. glareolusandA.flavi-collis, significant local differences as well as an effect of increasing body mass on cyst prevalence. Coccidian cystswere differentiated by amplification of the18S rRNAgene and subsequent sequencing. The majority of iden-tifiable cysts (97.9%) were determined asFrenkelia glareoli, a coccidian species mainly circulating betweenM.glareolusas intermediate and buzzards (Buteospp.) as definitive hosts. The zoonotic pathogenToxoplasma gondiiwas confirmed in oneM. glareolusoriginating from the city of Leipzig. Overall, it can be concluded that neu-roinvasion of zoonotic parasites seems to be rare inM. glareolusandA.flavicollis.
2019-10-21
2019-10-21
2019-12-01
Article
22132244
10.1016/j.ijppaw.2019.09.004
https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85072402445&origin=inward
http://hdl.handle.net/10033/621988
International Journal for Parasitology: Parasites and Wildlife
2-s2.0-85072402445
SCOPUS_ID:85072402445
S2213224419301476
en
International Journal for Parasitology: Parasites and Wildlife
10
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Elsevier
oai:repository.helmholtz-hzi.de:10033/6220252019-11-23T01:59:53Zcom_10033_6839col_10033_621495
The role of epigenetics in the development of childhood asthma.
Qi, Cancan
Xu, Cheng-Jian
Koppelman, Gerard H
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Asthma
DNA methylation
cell types
environmental exposures
epigenetics
Review
Introduction: The development of childhood asthma is caused by a combination of genetic factors and environmental exposures. Epigenetics describes mechanisms of (heritable) regulation of gene expression that occur without changes in DNA sequence. Epigenetics is strongly related to aging, is cell-type specific, and includes DNA methylation, noncoding RNAs, and histone modifications.Areas covered: This review summarizes recent epigenetic studies of childhood asthma in humans, which mostly involve studies of DNA methylation published in the recent five years. Environmental exposures, in particular cigarette smoking, have significant impact on epigenetic changes, but few of these epigenetic signals are also associated with asthma. Several asthma-associated genetic variants relate to DNA methylation. Epigenetic signals can be better understood by studying their correlation with gene expression, which revealed higher presence and activation of blood eosinophils in asthma. Strong associations of nasal methylation signatures and atopic asthma were identified, which were replicable across different populations.Expert commentary: Epigenetic markers have been strongly associated with asthma, and might serve as biomarker of asthma. The causal and longitudinal relationships between epigenetics and disease, and between environmental exposures and epigenetic changes need to be further investigated. Efforts should be made to understand cell-type-specific epigenetic mechanisms in asthma.
2019-11-22
2019-11-22
2019-11-10
Other
1744-8409
31674254
10.1080/1744666X.2020.1686977
http://hdl.handle.net/10033/622025
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
oai:repository.helmholtz-hzi.de:10033/6220432019-12-06T02:04:18Zcom_10033_6839col_10033_621495
Groundwater, soil and compost, as possible sources of virulent and antibiotic-resistant Pseudomonas aeruginosa.
Kaszab, Edit
Radó, Júlia
Kriszt, Balázs
Pászti, Judit
Lesinszki, Virág
Szabó, Ádám
Tóth, Gergő
Khaledi, Ariane
Szoboszlay, Sándor
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Pseudomonas aeruginosa
antibiotic resistance
biofilm
pulsed-field gel electrophoresis
virulence
Pseudomonas aeruginosa is a major public health concern all around the world. In the frame of this work, a set of diverse environmental P. aeruginosa isolates with various antibiotic resistance profiles were examined in a Galleria mellonella virulence model. Motility, serotypes, virulence factors and biofilm-forming ability were also examined. Molecular types were determined by pulsed-field gel electrophoresis (PFGE). Based on our results, the majority of environmental isolates were virulent in the G. mellonella test and twitching showed a positive correlation with mortality. Resistance against several antibiotic agents such as Imipenem correlated with a lower virulence in the applied G. mellonella model. PFGE revealed that five examined environmental isolates were closely related to clinically detected pulsed-field types. Our study demonstrated that industrial wastewater effluents, composts, and hydrocarbon-contaminated sites should be considered as hot spots of high-risk clones of P. aeruginosa.
2019-12-05
2019-12-05
2019-11-18
Article
Int J Environ Health Res. 2019 Nov 18:1-13. doi: 10.1080/09603123.2019.1691719.
1369-1619
31736330
10.1080/09603123.2019.1691719
http://hdl.handle.net/10033/622043
International Journal of Environmental Health Research
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Taylor & Francis
oai:repository.helmholtz-hzi.de:10033/6221132020-02-01T02:49:56Zcom_10033_6839col_10033_621495
Ex Vivo/In vivo Gene Editing in Hepatocytes Using "All-in-One" CRISPR-Adeno-Associated Virus Vectors with a Self-Linearizing Repair Template.
Krooss, Simon Alexander
Dai, Zhen
Schmidt, Florian
Rovai, Alice
Fakhiri, Julia
Dhingra, Akshay
Yuan, Qinggong
Yang, Taihua
Balakrishnan, Asha
Steinbrück, Lars
Srivaratharajan, Sangar
Manns, Michael Peter
Schambach, Axel
Grimm, Dirk
Bohne, Jens
Sharma, Amar Deep
Büning, Hildegard
Ott, Michael
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Genetic Engineering
Genetics
Techniques in Genetics
Adeno-associated virus (AAV)-based vectors are considered efficient and safe gene delivery systems in gene therapy. We combined two guide RNA genes, Cas9, and a self-linearizing repair template in one vector (AIO-SL) to correct fumarylacetoacetate hydrolase (FAH) deficiency in mice. The vector genome of 5.73 kb was packaged into VP2-depleted AAV particles (AAV2/8ΔVP2), which, however, did not improve cargo capacity. Reprogrammed hepatocytes were treated with AIO-SL.AAV2ΔVP2 and subsequently transplanted, resulting in large clusters of FAH-positive hepatocytes. Direct injection of AIO-SL.AAV8ΔVP2 likewise led to FAH expression and long-term survival. The AIO-SL vector achieved an ∼6-fold higher degree of template integration than vectors without template self-linearization. Subsequent analysis revealed that AAV8 particles, in contrast to AAV2, incorporate oversized genomes distinctly greater than 5.2 kb. Finally, our AAV8-based vector represents a promising tool for gene editing strategies to correct monogenic liver diseases requiring (large) fragment removal and/or simultaneous sequence replacement.
2020-01-31
2020-01-31
2020-01-24
Article
iScience. 2020 Jan 24;23(1):100764. doi: 10.1016/j.isci.2019.100764. Epub 2019 Dec 12.
2589-0042
31887661
10.1016/j.isci.2019.100764
http://hdl.handle.net/10033/622113
iScience
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Cell Press/Elsevier
oai:repository.helmholtz-hzi.de:10033/6221312020-02-13T02:08:59Zcom_10033_6839col_10033_621495
Direct recognition of hepatocyte-expressed MHC class I alloantigens is required for tolerance induction.
Paul-Heng, Moumita
Leong, Mario
Cunningham, Eithne
Bunker, Daniel L J
Bremner, Katherine
Wang, Zane
Wang, Chuanmin
Tay, Szun Szun
McGuffog, Claire
Logan, Grant J
Alexander, Ian E
Hu, Min
Alexander, Stephen I
Sparwasser, Tim D
Bertolino, Patrick
Bowen, David G
Bishop, G Alex
Sharland, Alexandra
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Antigen presentation
Gene therapy
Hepatology
Tolerance
Transplantation
Adeno-associated viral vector–mediated (AAV-mediated) expression of allogeneic major histocompatibility complex class I (MHC class I) in recipient liver induces donor-specific tolerance in mouse skin transplant models in which a class I allele (H-2Kb or H-2Kd) is mismatched between donor and recipient. Tolerance can be induced in mice primed by prior rejection of a donor-strain skin graft, as well as in naive recipients. Allogeneic MHC class I may be recognized by recipient T cells as an intact molecule (direct recognition) or may be processed and presented as an allogeneic peptide in the context of self-MHC (indirect recognition). The relative contributions of direct and indirect allorecognition to tolerance induction in this setting are unknown. Using hepatocyte-specific AAV vectors encoding WT allogeneic MHC class I molecules, or class I molecules containing a point mutation (D227K) that impedes direct recognition of intact allogeneic MHC class I by CD8+ T cells without hampering the presentation of processed peptides derived from allogeneic MHC class I, we show here that tolerance induction depends upon recognition of intact MHC class I. Indirect recognition alone yielded a modest prolongation of subsequent skin graft survival, attributable to the generation of CD4+ Tregs, but it was not sufficient to induce tolerance.
2020-02-12
2020-02-12
2018-08-09
Article
JCI Insight. 2018 Aug 9;3(15). pii: 97500. doi: 10.1172/jci.insight.97500. eCollection 2018 Aug 9.
2379-3708
30089715
10.1172/jci.insight.97500
http://hdl.handle.net/10033/622131
JCI Insight
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
NLM (Medline)
oai:repository.helmholtz-hzi.de:10033/6222392020-04-22T04:53:58Zcom_10033_6839col_10033_621495
TLR4 abrogates the Th1 immune response through IRF1 and IFN-β to prevent immunopathology during L. infantum infection.
Sacramento, Laís Amorim
Benevides, Luciana
Maruyama, Sandra Regina
Tavares, Lucas
Fukutani, Kiyoshi Ferreira
Francozo, Marcela
Sparwasser, Tim
Cunha, Fernando Queiroz
Almeida, Roque Pacheco
da Silva, João Santana
Carregaro, Vanessa
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
A striking feature of human visceral leishmaniasis (VL) is chronic inflammation in the spleen and liver, and VL patients present increased production levels of multiple inflammatory mediators, which contribute to tissue damage and disease severity. Here, we combined an experimental model with the transcriptional profile of human VL to demonstrate that the TLR4-IFN-β pathway regulates the chronic inflammatory process and is associated with the asymptomatic form of the disease. Tlr4-deficient mice harbored fewer parasites in their spleen and liver than wild-type mice. TLR4 deficiency enhanced the Th1 immune response against the parasite, which was correlated with an increased activation of dendritic cells (DCs). Gene expression analyses demonstrated that IRF1 and IFN-β were expressed downstream of TLR4 after infection. Accordingly, IRF1- and IFNAR-deficient mice harbored fewer parasites in the target organs than wild-type mice due to having an increased Th1 immune response. However, the absence of TLR4 or IFNAR increased the serum transaminase levels in infected mice, indicating the presence of liver damage in these animals. In addition, IFN-β limits IFN-γ production by acting directly on Th1 cells. Using RNA sequencing analysis of human samples, we demonstrated that the transcriptional signature for the TLR4 and type I IFN (IFN-I) pathways was positively modulated in asymptomatic subjects compared with VL patients and thus provide direct evidence demonstrating that the TLR4-IFN-I pathway is related to the nondevelopment of the disease. In conclusion, our results demonstrate that the TLR4-IRF1 pathway culminates in IFN-β production as a mechanism for dampening the chronic inflammatory process and preventing immunopathology development.
2020-04-21
2020-04-21
2020-03-25
Article
PLoS Pathog. 2020 Mar 25;16(3):e1008435. doi: 10.1371/journal.ppat.1008435. eCollection 2020 Mar.
32210480
10.1371/journal.ppat.1008435
http://hdl.handle.net/10033/622239
1553-7374
PLoS pathogens
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
PLOS
oai:repository.helmholtz-hzi.de:10033/6222992020-06-17T01:30:44Zcom_10033_6839col_10033_621495
The Dynamics of Interleukin-10-Afforded Protection during Dextran Sulfate Sodium-Induced Colitis.
Cardoso, Ana
Gil Castro, Antonio
Martins, Ana Catarina
Carriche, Guilhermina M
Murigneux, Valentine
Castro, Isabel
Cumano, Ana
Vieira, Paulo
Saraiva, Margarida
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
colitis
inflammation
interleukin-10
macrophages
therapy
Inflammatory bowel disease encompasses a group of chronic-inflammatory conditions of the colon and small intestine. These conditions are characterized by exacerbated inflammation of the organ that greatly affects the quality of life of patients. Molecular mechanisms counteracting this hyperinflammatory status of the gut offer strategies for therapeutic intervention. Among these regulatory molecules is the anti-inflammatory cytokine interleukin (IL)-10, as shown in mice and humans. Indeed, IL-10 signaling, particularly in macrophages, is essential for intestinal homeostasis. We sought to investigate the temporal profile of IL-10-mediated protection during chemical colitis and which were the underlying mechanisms. Using a novel mouse model of inducible IL-10 overexpression (pMT-10), described here, we show that mice preconditioned with IL-10 for 8 days before dextran sulfate sodium (DSS) administration developed a milder colitic phenotype. In IL-10-induced colitic mice, Ly6C cells isolated from the lamina propria showed a decreased inflammatory profile. Because our mouse model leads to transcription of the IL-10 transgene in the bone marrow and elevated seric IL-10 concentration, we investigated whether IL-10 could imprint immune cells in a long-lasting way, thus conferring sustained protection to colitis. We show that this was not the case, as IL-10-afforded protection was only observed if IL-10 induction immediately preceded DSS-mediated colitis. Thus, despite the protection afforded by IL-10 in colitis, novel strategies are required, specifically to achieve long-lasting protection.
2020-06-16
2020-06-16
2018-03-01
Article
Other
Front Immunol. 2018;9:400. Published 2018 Mar 1. doi:10.3389/fimmu.2018.00400.
1664-3224
29545807
10.3389/fimmu.2018.00400
http://hdl.handle.net/10033/622299
Frontiers in immunology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Frontiers
oai:repository.helmholtz-hzi.de:10033/6223132020-06-26T02:39:20Zcom_10033_6839col_10033_621495
Let-7c inhibits cholangiocarcinoma growth but promotes tumor cell invasion and growth at extrahepatic sites.
Xie, Yu
Zhang, Hang
Guo, Xing-Jun
Feng, Ye-Chen
He, Rui-Zhi
Li, Xu
Yu, Shuo
Zhao, Yan
Shen, Ming
Zhu, Feng
Wang, Xin
Wang, Min
Balakrishnan, Asha
Ott, Michael
Peng, Feng
Qin, Ren-Yi
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Cholangiocarcinoma (CCA) is a cancer type with high postoperative relapse rates and poor long-term survival largely due to tumor invasion, distant metastasis, and multidrug resistance. Deregulated microRNAs (miRNAs) are implicated in several cancer types including CCA. The specific roles of the miRNA let-7c in cholangiocarcinoma are not known and need to be further elucidated. In our translational study we show that microRNA let-7c expression was significantly downregulated in human cholangiocarcinoma tissues when compared to adjacent tissues of the same patient. Let-7c inhibited the tumorigenic properties of cholangiocarcinoma cells including their self-renewal capacity and sphere formation in vitro and subcutaneous cancer cell growth in vivo. Ectopic let-7c overexpression suppressed migration and invasion capacities of cholangiocarcinoma cell lines in vitro, however, promoted distant invasiveness in vivo. Furthermore, we found that let-7c regulated the aforementioned malignant biological properties, at least in part, through regulation of EZH2 protein expression and through the DVL3/β-catenin axis. The miRNA let-7c thus plays an important dual role in regulating tumorigenic and metastatic abilities of human cholangiocarcinoma through mechanisms involving EZH2 protein and the DVL3/β-catenin axis.
2020-06-25
2020-06-25
2018-02-14
Article
Other
29445149
10.1038/s41419-018-0286-6
http://hdl.handle.net/10033/622313
2041-4889
Cell death & disease
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Springer Nature
oai:repository.helmholtz-hzi.de:10033/6223952020-08-08T02:31:43Zcom_10033_6839col_10033_621495
Pathological mechanism and antisense oligonucleotide-mediated rescue of a non-coding variant suppressing factor 9 RNA biogenesis leading to hemophilia B.
Krooss, Simon
Werwitzke, Sonja
Kopp, Johannes
Rovai, Alice
Varnholt, Dirk
Wachs, Amelie S
Goyenvalle, Aurelie
Aarstma-Rus, Annemieke
Ott, Michael
Tiede, Andreas
Langemeier, Jörg
Bohne, Jens
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Loss-of-function mutations in the human coagulation factor 9 (F9) gene lead to hemophilia B. Here, we dissected the consequences and the pathomechanism of a non-coding mutation (c.2545A>G) in the F9 3' untranslated region. Using wild type and mutant factor IX (FIX) minigenes we revealed that the mutation leads to reduced F9 mRNA and FIX protein levels and to lower coagulation activity of cell culture supernatants. The phenotype could not be compensated by increased transcription. The pathomechanism comprises the de novo creation of a binding site for the spliceosomal component U1snRNP, which is able to suppress the nearby F9 poly(A) site. This second, splicing-independent function of U1snRNP was discovered previously and blockade of U1snRNP restored mutant F9 mRNA expression. In addition, we explored the vice versa approach and masked the mutation by antisense oligonucleotides resulting in significantly increased F9 mRNA expression and coagulation activity. This treatment may transform the moderate/severe hemophilia B into a mild or subclinical form in the patients. This antisense based strategy is applicable to other mutations in untranslated regions creating deleterious binding sites for cellular proteins.
2020-08-07
2020-08-07
2020-04-08
Article
Other
PLoS Genet. 2020;16(4):e1008690. Published 2020 Apr 8. doi:10.1371/journal.pgen.1008690.
32267853
10.1371/journal.pgen.1008690
http://hdl.handle.net/10033/622395
1553-7404
PLoS genetics
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
PLOS
oai:repository.helmholtz-hzi.de:10033/6224102020-08-18T01:29:44Zcom_10033_6839col_10033_621495
Establishment of porcine and human expanded potential stem cells.
Gao, Xuefei
Nowak-Imialek, Monika
Chen, Xi
Chen, Dongsheng
Herrmann, Doris
Ruan, Degong
Chen, Andy Chun Hang
Eckersley-Maslin, Melanie A
Ahmad, Shakil
Lee, Yin Lau
Kobayashi, Toshihiro
Ryan, David
Zhong, Jixing
Zhu, Jiacheng
Wu, Jian
Lan, Guocheng
Petkov, Stoyan
Yang, Jian
Antunes, Liliana
Campos, Lia S
Fu, Beiyuan
Wang, Shengpeng
Yong, Yu
Wang, Xiaomin
Xue, Song-Guo
Ge, Liangpeng
Liu, Zuohua
Huang, Yong
Nie, Tao
Li, Peng
Wu, Donghai
Pei, Duanqing
Zhang, Yi
Lu, Liming
Yang, Fengtang
Kimber, Susan J
Reik, Wolf
Zou, Xiangang
Shang, Zhouchun
Lai, Liangxue
Surani, Azim
Tam, Patrick P L
Ahmed, Asif
Yeung, William Shu Biu
Teichmann, Sarah A
Niemann, Heiner
Liu, Pentao
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
We recently derived mouse expanded potential stem cells (EPSCs) from individual blastomeres by inhibiting the critical molecular pathways that predispose their differentiation. EPSCs had enriched molecular signatures of blastomeres and possessed developmental potency for all embryonic and extra-embryonic cell lineages. Here, we report the derivation of porcine EPSCs, which express key pluripotency genes, are genetically stable, permit genome editing, differentiate to derivatives of the three germ layers in chimeras and produce primordial germ cell-like cells in vitro. Under similar conditions, human embryonic stem cells and induced pluripotent stem cells can be converted, or somatic cells directly reprogrammed, to EPSCs that display the molecular and functional attributes reminiscent of porcine EPSCs. Importantly, trophoblast stem-cell-like cells can be generated from both human and porcine EPSCs. Our pathway-inhibition paradigm thus opens an avenue for generating mammalian pluripotent stem cells, and EPSCs present a unique cellular platform for translational research in biotechnology and regenerative medicine.
2020-08-17
2020-08-17
2019-06-03
Article
Other
Nat Cell Biol. 2019;21(6):687-699. doi:10.1038/s41556-019-0333-2.
31160711
10.1038/s41556-019-0333-2
http://hdl.handle.net/10033/622410
1476-4679
Nature cell biology
PMC7035105
en
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7035105/
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Nature publishing group(NPG)
oai:repository.helmholtz-hzi.de:10033/6224332020-09-28T10:51:11Zcom_10033_128109com_10033_620659com_10033_6839com_10033_620618col_10033_128110col_10033_620660col_10033_621495col_10033_620621
Staphylococcus aureus Alpha-Toxin Limits Type 1 While Fostering Type 3 Immune Responses.
Bonifacius, Agnes
Goldmann, Oliver
Floess, Stefan
Holtfreter, Silva
Robert, Philippe A
Nordengrün, Maria
Kruse, Friederike
Lochner, Matthias
Falk, Christine S
Schmitz, Ingo
Bröker, Barbara M
Medina, Eva
Huehn, Jochen
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
CD4+ T cells
Staphylococcus aureus
alpha-toxin
innate lymphoid cells
γδ T cells
Staphylococcus aureus can cause life-threatening diseases, and hospital- as well as community-associated antibiotic-resistant strains are an emerging global public health problem. Therefore, prophylactic vaccines or immune-based therapies are considered as alternative treatment opportunities. To develop such novel treatment approaches, a better understanding of the bacterial virulence and immune evasion mechanisms and their potential effects on immune-based therapies is essential. One important staphylococcal virulence factor is alpha-toxin, which is able to disrupt the epithelial barrier in order to establish infection. In addition, alpha-toxin has been reported to modulate other cell types including immune cells. Since CD4+ T cell-mediated immunity is required for protection against S. aureus infection, we were interested in the ability of alpha-toxin to directly modulate CD4+ T cells. To address this, murine naïve CD4+ T cells were differentiated in vitro into effector T cell subsets in the presence of alpha-toxin. Interestingly, alpha-toxin induced death of Th1-polarized cells, while cells polarized under Th17 conditions showed a high resistance toward increasing concentrations of this toxin. These effects could neither be explained by differential expression of the cellular alpha-toxin receptor ADAM10 nor by differential activation of caspases, but might result from an increased susceptibility of Th1 cells toward Ca2+-mediated activation-induced cell death. In accordance with the in vitro findings, an alpha-toxin-dependent decrease of Th1 and concomitant increase of Th17 cells was observed in vivo during S. aureus bacteremia. Interestingly, corresponding subsets of innate lymphoid cells and γδ T cells were similarly affected, suggesting a more general effect of alpha-toxin on the modulation of type 1 and type 3 immune responses. In conclusion, we have identified a novel alpha-toxin-dependent immunomodulatory strategy of S. aureus, which can directly act on CD4+ T cells and might be exploited for the development of novel immune-based therapeutic approaches to treat infections with antibiotic-resistant S. aureus strains.
2020-09-10
2020-09-10
2020-08-07
Article
Front Immunol. 2020;11:1579. Published 2020 Aug 7. doi:10.3389/fimmu.2020.01579.
32849537
10.3389/fimmu.2020.01579
http://hdl.handle.net/10033/622433
1664-3224
Frontiers in immunology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Frontiers
oai:repository.helmholtz-hzi.de:10033/6224582020-09-25T03:25:09Zcom_10033_6839col_10033_621495
Strategic Anti-SARS-CoV-2 Serology Testing in a Low Prevalence Setting: The COVID-19 Contact (CoCo) Study in Healthcare Professionals.
Behrens, Georg M N
Cossmann, Anne
Stankov, Metodi V
Schulte, Bianca
Streeck, Hendrik
Förster, Reinhold
Bosnjak, Berislav
Willenzon, Stefanie
Boeck, Anna-Lena
Thu Tran, Anh
Thiele, Thea
Graalmann, Theresa
Kayser, Moritz Z
Zychlinsky Scharff, Anna
Dopfer, Christian
Horke, Alexander
Pink, Isabell
Witte, Torsten
Wetzke, Martin
Ernst, Diana
Jablonka, Alexandra
Happle, Christine
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
COVID-19
Coronavirus
Healthcare professionals
Humoral immunity
Infection
Pandemic
SARS-CoV-2
Serological testing
Virus
Background: Serology testing is explored for epidemiological research and to inform individuals after suspected infection. During the coronavirus disease 2019 (COVID-19) pandemic, frontline healthcare professionals (HCP) may be at particular risk for infection. No longitudinal data on functional seroconversion in HCP in regions with low COVID-19 prevalence and low pre-test probability exist.
Methods: In a large German university hospital, we performed weekly questionnaire assessments and anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immunoglobulin G (IgG) measurements with various commercial tests, a novel surrogate virus neutralisation test, and a neutralisation assay using live SARS-CoV-2.
Results: From baseline to week 6, 1080 screening measurements for anti-SARS CoV-2 (S1) IgG from 217 frontline HCP (65% female) were performed. Overall, 75.6% of HCP reported at least one symptom of respiratory infection. Self-perceived infection probability declined over time (from mean 20.1% at baseline to 12.4% in week 6, p < 0.001). In sera of convalescent patients with PCR-confirmed COVID-19, we measured high anti-SARS-CoV-2 IgG levels, obtained highly concordant results from enzyme-linked immunosorbent assays (ELISA) using e.g. the spike 1 (S1) protein domain and the nucleocapsid protein (NCP) as targets, and confirmed antiviral neutralisation. However, in HCP the cumulative incidence for anti-SARS-CoV-2 (S1) IgG was 1.86% for positive and 0.93% for equivocal positive results over the study period of 6 weeks. Except for one HCP, none of the eight initial positive results were confirmed by alternative serology tests or showed in vitro neutralisation against live SARS-CoV-2. The only true seroconversion occurred without symptoms and mounted strong functional humoral immunity. Thus, the confirmed cumulative incidence for neutralizing anti-SARS-CoV-2 IgG was 0.47%.
Conclusion: When assessing anti-SARS-CoV-2 immune status in individuals with low pre-test probability, we suggest confirming positive results from single measurements by alternative serology tests or functional assays. Our data highlight the need for a methodical serology screening approach in regions with low SARS-CoV-2 infection rates.
2020-09-24
2020-09-24
2020-09-04
Article
nfect Dis Ther. 2020 Sep 4:1–13. doi: 10.1007/s40121-020-00334-1. Epub ahead of print. PMID: 32886335.
2193-8229
32886335
10.1007/s40121-020-00334-1
http://hdl.handle.net/10033/622458
Infectious diseases and therapy
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Springer Healthcare
oai:repository.helmholtz-hzi.de:10033/6225872020-11-17T01:36:41Zcom_10033_6839col_10033_621495
Notch and TLR signaling coordinate monocyte cell fate and inflammation.
Gamrekelashvili, Jaba
Kapanadze, Tamar
Sablotny, Stefan
Ratiu, Corina
Dastagir, Khaled
Lochner, Matthias
Karbach, Susanne
Wenzel, Philip
Sitnow, Andre
Fleig, Susanne
Sparwasser, Tim
Kalinke, Ulrich
Holzmann, Bernhard
Haller, Hermann
Limbourg, Florian P
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
TLR signaling
immunology
inflammation
macrophage differentiation
monocytes and macrophages
mouse
notch signaling
Conventional Ly6Chi monocytes have developmental plasticity for a spectrum of differentiated phagocytes. Here we show, using conditional deletion strategies in a mouse model of Toll-like receptor (TLR) 7-induced inflammation, that the spectrum of developmental cell fates of Ly6Chi monocytes, and the resultant inflammation, is coordinately regulated by TLR and Notch signaling. Cell-intrinsic Notch2 and TLR7-Myd88 pathways independently and synergistically promote Ly6Clo patrolling monocyte development from Ly6Chi monocytes under inflammatory conditions, while impairment in either signaling axis impairs Ly6Clo monocyte development. At the same time, TLR7 stimulation in the absence of functional Notch2 signaling promotes resident tissue macrophage gene expression signatures in monocytes in the blood and ectopic differentiation of Ly6Chi monocytes into macrophages and dendritic cells, which infiltrate the spleen and major blood vessels and are accompanied by aberrant systemic inflammation. Thus, Notch2 is a master regulator of Ly6Chi monocyte cell fate and inflammation in response to TLR signaling.
2020-11-16
2020-11-16
2020-07-29
Article
Elife. 2020 Jul 29;9:e57007. doi: 10.7554/eLife.57007.
32723480
10.7554/eLife.57007
http://hdl.handle.net/10033/622587
2050-084X
eLife
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
elife Sciences
oai:repository.helmholtz-hzi.de:10033/6225982020-11-24T09:46:05Zcom_10033_6839col_10033_621495
MicroRNA-221: A Fine Tuner and Potential Biomarker of Chronic Liver Injury.
Markovic, Jovana
Sharma, Amar Deep
Balakrishnan, Asha
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
HCC
NASH
liver fibrosis
miR-221
noninvasive biomarker
The last decade has witnessed significant advancements in our understanding of how small noncoding RNAs, such as microRNAs (miRNAs), regulate disease progression. One such miRNA, miR-221, has been shown to play a key role in the progression of liver fibrosis, a common feature of most liver diseases. Many reports have demonstrated the upregulation of miR-221 in liver fibrosis caused by multiple etiologies such as viral infections and nonalcoholic steatohepatitis. Inhibition of miR-221 via different strategies has shown promising results in terms of the suppression of fibrogenic gene signatures in vitro, as well as in vivo, in independent mouse models of liver fibrosis. In addition, miR-221 has also been suggested as a noninvasive serum biomarker for liver fibrosis and cirrhosis. In this review, we discuss the biology of miR-221, its significance and use as a biomarker during progression of liver fibrosis, and finally, potential and robust approaches that can be utilized to suppress liver fibrosis via inhibition of miR-221.
2020-11-19
2020-11-19
2020-07-23
Review
Cells. 2020 Jul 23;9(8):1767. doi: 10.3390/cells9081767.
32717951
10.3390/cells9081767
http://hdl.handle.net/10033/622598
2073-4409
Cells
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
MDPI
oai:repository.helmholtz-hzi.de:10033/6226282021-08-12T09:20:57Zcom_10033_6839col_10033_621495
MicroRNA-342-3p is a potent tumour suppressor in hepatocellular carcinoma
Komoll, Ronja Melinda
Hu, Qingluan
Olarewaju, Olaniyi
von Döhlen, Lena
Yuan, Qinggong
Xie, Yu
Tsay, Hsin Chieh
Daon, Joel
Qin, Renyi
Manns, Michael P
Sharma, Amar Deep
Goga, Andrei
Ott, Michael
Balakrishnan, Asha
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Hepatocellular carcinoma
Lactate transport
Liver cancer
MCT1
MicroRNAs
MYC
RAS
Tumour metabolism
Tumour regression
Background & aims: Hepatocellular carcinoma (HCC) is a cancer with multiple aetiologies and widespread prevalence. Largely refractory to current treatments, HCC is the fourth leading cause of cancer-related deaths worldwide. MicroRNAs (miRNAs) are important regulators in HCCs. We aimed to identify tumour suppressor miRNAs during tumour regression in a conditional c-MYC-driven mouse model (LT2/MYC) of HCC, and to evaluate their therapeutic potential for HCC treatment.
Methods: We performed miRNA expression profiling of developed and regressing LT2/MYC tumours and in-depth in vitro gain- and loss-of-function analyses. The effect of adeno-associated virus (AAV) vector-mediated miR-342-3p treatment was evaluated in 3 HCC mouse models.
Results: We identified miR-342-3p as a tumour suppressor miRNA in HCC, with increased expression in regressing tumours. Forced miR-342-3p expression in hepatoma cells showed significantly decreased cell proliferation, migration, and colony formation. In vivo administration of AAV-miR-342-3p led to significant attenuation of tumour development and increased overall survival. We identified monocarboxylic acid transporter 1 (MCT1) as a bona fide target of miR-342-3p in HCC. We show that the tumour suppressor role of miR-342-3p is executed partly by modulating the lactate transport function of MCT1. Importantly, we find miR-342-3p downregulated in tumours from patients with HCC compared with matched non-tumour tissues, inversely correlating with MCT1 expression. We observed similar findings in TCGA-LIHC data.
Conclusions: In our study, we identified and validated miR-342-3p as a tumour suppressor miRNA in HCC. We demonstrated its therapeutic efficacy in significantly attenuating tumour development, and prolonging survival, in different HCC mouse models. Identification of miR-342-3p as an effective tumour suppressor opens a therapeutic avenue for miRNA-mediated attenuation of HCC development.
Lay summary: Hepatocellular carcinoma (HCC), the most common type of liver cancer, affects diverse populations and has a global impact, being the fourth leading cause of cancer deaths worldwide. There are currently no systemic therapies for HCC that can significantly prolong long-term survival. Thus, novel effective treatment options are urgently required. To understand the molecular basis of tumour regression, we compared tumours and regressing liver tumours in mice. We show that a small non-coding miRNA, miR-342-3p, is a tumour suppressor in HCC. Expression of miR-342-3p is low in tumours and high in regressing tumours. When miR-342-3p is delivered to mouse livers with HCC, it can significantly slow down liver tumour development and improve survival. Our study highlights the promising therapeutic potential of miR-342-3p intervention in HCC.
2020-12-02
2020-12-02
2021-01-01
Article
J Hepatol. 2020 Jul 30:S0168-8278(20)30492-X. doi: 10.1016/j.jhep.2020.07.039. Epub ahead of print.
01688278
10.1016/j.jhep.2020.07.039
http://hdl.handle.net/10033/622628
16000641
Journal of Hepatology
2-s2.0-85093503509
SCOPUS_ID:85093503509
S016882782030492X
en
http://creativecommons.org/licenses/by-nc/4.0/
Attribution-NonCommercial 4.0 International
oai:repository.helmholtz-hzi.de:10033/6226392020-12-10T01:43:33Zcom_10033_6839col_10033_621495
MicroRNA-342-3p is a potent tumour suppressor in hepatocellular carcinoma.
Komoll, Ronja-Melinda
Hu, Qingluan
Olarewaju, Olaniyi
von Döhlen, Lena
Yuan, Qinggong
Xie, Yu
Tsay, Hsin-Chieh
Daon, Joel
Qin, Renyi
Manns, Michael P
Sharma, Amar Deep
Goga, Andrei
Ott, Michael
Balakrishnan, Asha
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Hepatocellular carcinoma
Lactate transport
Liver cancer
MCT1
MYC
MicroRNAs
RAS
Tumour metabolism
Tumour regression
Background & aims: Hepatocellular carcinoma (HCC) is a cancer with multiple aetiologies and widespread prevalence. Largely refractory to current treatments, HCC is the fourth leading cause of cancer-related deaths worldwide. MicroRNAs (miRNAs) are important regulators in HCCs. We aimed to identify tumour suppressor miRNAs during tumour regression in a conditional c-MYC-driven mouse model (LT2/MYC) of HCC, and to evaluate their therapeutic potential for HCC treatment.
Methods: We performed miRNA expression profiling of developed and regressing LT2/MYC tumours and in-depth in vitro gain- and loss-of-function analyses. The effect of adeno-associated virus (AAV) vector-mediated miR-342-3p treatment was evaluated in 3 HCC mouse models.
Results: We identified miR-342-3p as a tumour suppressor miRNA in HCC, with increased expression in regressing tumours. Forced miR-342-3p expression in hepatoma cells showed significantly decreased cell proliferation, migration, and colony formation. In vivo administration of AAV-miR-342-3p led to significant attenuation of tumour development and increased overall survival. We identified monocarboxylic acid transporter 1 (MCT1) as a bona fide target of miR-342-3p in HCC. We show that the tumour suppressor role of miR-342-3p is executed partly by modulating the lactate transport function of MCT1. Importantly, we find miR-342-3p downregulated in tumours from patients with HCC compared with matched non-tumour tissues, inversely correlating with MCT1 expression. We observed similar findings in TCGA-LIHC data.
Conclusions: In our study, we identified and validated miR-342-3p as a tumour suppressor miRNA in HCC. We demonstrated its therapeutic efficacy in significantly attenuating tumour development, and prolonging survival, in different HCC mouse models. Identification of miR-342-3p as an effective tumour suppressor opens a therapeutic avenue for miRNA-mediated attenuation of HCC development.
Lay summary: Hepatocellular carcinoma (HCC), the most common type of liver cancer, affects diverse populations and has a global impact, being the fourth leading cause of cancer deaths worldwide. There are currently no systemic therapies for HCC that can significantly prolong long-term survival. Thus, novel effective treatment options are urgently required. To understand the molecular basis of tumour regression, we compared tumours and regressing liver tumours in mice. We show that a small non-coding miRNA, miR-342-3p, is a tumour suppressor in HCC. Expression of miR-342-3p is low in tumours and high in regressing tumours. When miR-342-3p is delivered to mouse livers with HCC, it can significantly slow down liver tumour development and improve survival. Our study highlights the promising therapeutic potential of miR-342-3p intervention in HCC.
2020-12-09
2020-12-09
2020-07-30
Article
J Hepatol. 2020 Jul 30:S0168-8278(20)30492-X. doi: 10.1016/j.jhep.2020.07.039. Epub
32738449
10.1016/j.jhep.2020.07.039
http://hdl.handle.net/10033/622639
1600-0641
Journal of hepatology
en
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Elsevier
oai:repository.helmholtz-hzi.de:10033/6226462020-12-12T10:30:59Zcom_10033_6839col_10033_621495
Cholesterol sensing by CD81 is important for hepatitis C virus entry.
Palor, Machaela
Stejskal, Lenka
Mandal, Piya
Lenman, Annasara
Alberione, Maria Pia
Kirui, Jared
Moeller, Rebecca
Ebner, Stefan
Meissner, Felix
Gerold, Gisa
Shepherd, Adrian J
Grove, Joe
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Hepatitis C virus (HCV)
cholesterol-binding protein
molecular dynamics
tetraspanin
virus entry
CD81 plays a role in a variety of physiological and pathological processes. Recent structural analysis of CD81 indicates that it contains an intramembrane cholesterol-binding pocket and that interaction with cholesterol may regulate a conformational switch in the extracellular domain of CD81. Therefore, CD81 possesses a potential cholesterol sensing mechanism; however, its relevance for protein function is thus far unknown. In this study we investigate CD81 cholesterol sensing in the context of its activity as a receptor for hepatitis C virus. Structure-led mutagenesis of the cholesterol-binding pocket reduced CD81-cholesterol association, but had disparate effects on HCV, both reducing and enhancing CD81 receptor activity. We reasoned that this could be explained by alterations in the consequences of cholesterol binding. To investigate this further we performed molecular dynamic simulations of CD81 with and without cholesterol; this identified an allosteric mechanism by which cholesterol binding regulates the conformation of CD81. To test this, we designed further mutations to force CD81 into either the open (cholesterol unbound) or closed (cholesterol bound) conformation. The open mutant of CD81 exhibited reduced receptor activity whereas the closed mutant was enhanced. These data are consistent with cholesterol switching CD81 between a receptor active and inactive state. CD81 interactome analysis also suggests that conformational switching may modulate the assembly of CD81-partner networks. This work furthers our understanding of the molecular mechanism of CD81 cholesterol sensing, how this relates to HCV entry and CD81's function as a molecular scaffold; these insights are relevant to CD81's varied roles in health and disease.
2020-12-11
2020-12-11
2020-09-08
Article
J Biol Chem. 2020 Sep 8:jbc.RA120.014761. doi: 10.1074/jbc.RA120.014761. Epub ahead of print.
32900848
10.1074/jbc.RA120.014761
http://hdl.handle.net/10033/622646
1083-351X
The Journal of biological chemistry
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
DeGruyter
oai:repository.helmholtz-hzi.de:10033/6227112021-02-03T01:34:19Zcom_10033_6839col_10033_621495
HBV evolution and genetic variability: Impact on prevention, treatment and development of antivirals.
Glebe, Dieter
Goldmann, Nora
Lauber, Chris
Seitz, Stefan
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Antiviral treatment
HBV genotype diversity
HBV vaccination
Hepadnaviruses
Hepatitis B virus
Virus evolution
Hepatitis B virus (HBV) poses a major global health burden with 260 million people being chronically infected and 890,000 dying annually from complications in the course of the infection. HBV is a small enveloped virus with a reverse-transcribed DNA genome that infects hepatocytes and can cause acute and chronic infections of the liver. HBV is endemic in humans and apes representing the prototype member of the viral family Hepadnaviridae and can be divided into 10 genotypes. Hepadnaviruses have been found in all vertebrate classes and constitute an ancient viral family that descended from non-enveloped progenitors more than 360 million years ago. The de novo emergence of the envelope protein gene was accompanied with the liver-tropism and resulted in a tight virus-host association. The oldest HBV genomes so far have been isolated from human remains of the Bronze Age and the Neolithic (~7000 years before present). Despite the remarkable stability of the hepadnaviral genome over geological eras, HBV is able to rapidly evolve within an infected individual under pressure of the immune response or during antiviral treatment. Treatment with currently available antivirals blocking intracellular replication of HBV allows controlling of high viremia and improving liver health during long-term therapy of patients with chronic hepatitis B (CHB), but they are not sufficient to cure the disease. New therapy options that cover all HBV genotypes and emerging viral variants will have to be developed soon. In addition to the antiviral treatment of chronically infected patients, continued efforts to expand the global coverage of the currently available HBV vaccine will be one of the key factors for controlling the rising global spread of HBV. Certain improvements of the vaccine (e.g. inclusion of PreS domains) could counteract known problems such as low or no responsiveness of certain risk groups and waning anti-HBs titers leading to occult infections, especially with HBV genotypes E or F. But even with an optimal vaccine and a cure for hepatitis B, global eradication of HBV would be difficult to achieve because of an existing viral reservoir in primates and bats carrying closely related hepadnaviruses with zoonotic potential.
2021-02-02
2021-02-02
2020-11-06
Article
Antiviral Res. 2020 Nov 6;186:104973. doi: 10.1016/j.antiviral.2020.104973. Epub ahead of print.
33166575
10.1016/j.antiviral.2020.104973
http://hdl.handle.net/10033/622711
1872-9096
Antiviral research
en
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Elsevier
oai:repository.helmholtz-hzi.de:10033/6227662021-03-04T03:35:51Zcom_10033_6839col_10033_621495
Regulatory T Cells in an Endogenous Mouse Lymphoma Recognize Specific Antigen Peptides and Contribute to Immune Escape.
Ahmetlić, Fatima
Riedel, Tanja
Hömberg, Nadine
Bauer, Vera
Trautwein, Nico
Geishauser, Albert
Sparwasser, Tim
Stevanović, Stefan
Röcken, Martin
Mocikat, Ralph
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Foxp3+ regulatory T cells (Tregs) sustain immune homeostasis and may contribute to immune escape in malignant disease. As a prerequisite for developing immunologic approaches in cancer therapy, it is necessary to understand the ontogeny and the antigenic specificities of tumor-infiltrating Tregs. We addressed this question by using a λ-MYC transgenic mouse model of endogenously arising B-cell lymphoma, which mirrors key features of human Burkitt lymphoma. We show that Foxp3+ Tregs suppress antitumor responses in endogenous lymphoma. Ablation of Foxp3+ Tregs significantly delayed tumor development. The ratio of Treg to effector T cells was elevated in growing tumors, which could be ascribed to differential proliferation. The Tregs detected were mainly natural Tregs that apparently recognized self-antigens. We identified MHC class II-restricted nonmutated self-epitopes, which were more prevalent in lymphoma than in normal B cells and could be recognized by Tregs. These epitopes were derived from proteins that are associated with cellular processes related to malignancy and may be overexpressed in the tumor.
2021-03-03
2021-03-03
2019-03-20
Article
Cancer Immunol Res. 2019 Apr;7(4):600-608. doi: 10.1158/2326-6066.CIR-18-0419. Epub 2019 Mar 20.
30894379
10.1158/2326-6066.CIR-18-0419
http://hdl.handle.net/10033/622766
2326-6074
Cancer immunology research
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
American Association for Cancer Research (AACR)
oai:repository.helmholtz-hzi.de:10033/6227742021-03-06T01:38:46Zcom_10033_6839col_10033_621495
Selective Host Cell Death by Staphylococcus aureus : A Strategy for Bacterial Persistence.
Missiakas, Dominique
Winstel, Volker
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Staphylococcus aureus
abscess
blood stream infection
host cell death
infection
persistence
Host cell death programs are fundamental processes that shape cellular homeostasis, embryonic development, and tissue regeneration. Death signaling and downstream host cell responses are not only critical to guide mammalian development, they often act as terminal responses to invading pathogens. Here, we briefly review and contrast how invading pathogens and specifically Staphylococcus aureus manipulate apoptotic, necroptotic, and pyroptotic cell death modes to establish infection. Rather than invading host cells, S. aureus subverts these cells to produce diffusible molecules that cause death of neighboring hematopoietic cells and thus shapes an immune environment conducive to persistence. The exploitation of cell death pathways by S. aureus is yet another virulence strategy that must be juxtaposed to mechanisms of immune evasion, autophagy escape, and tolerance to intracellular killing, and brings us closer to the true portrait of this pathogen for the design of effective therapeutics and intervention strategies.
2021-03-05
2021-03-05
2021-01-21
Article
Front Immunol. 2021 Jan 21;11:621733. doi: 10.3389/fimmu.2020.621733.
33552085
10.3389/fimmu.2020.621733
http://hdl.handle.net/10033/622774
1664-3224
Frontiers in immunology
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
Frontiers
oai:repository.helmholtz-hzi.de:10033/6228042021-03-31T01:32:13Zcom_10033_6839col_10033_621495
Clarithromycin impairs tissue-resident memory and Th17 responses to macrolide-resistant Streptococcus pneumoniae infections.
Lindenberg, Marc
Almeida, Luis
Dhillon-LaBrooy, Ayesha
Siegel, Ekkehard
Henriques-Normark, Birgitta
Sparwasser, Tim
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Anti-microbial resistance
Clarithromycin
Macrolide antibiotics
Streptococcus pneumoniae
Th17 cells
Tissue-resident memory T cells
The increasing prevalence of antimicrobial resistance in pathogens is a growing public health concern, with the potential to compromise the success of infectious disease treatments in the future. Particularly, the number of infections by macrolide antibiotics-resistant Streptococcus pneumoniae is increasing. We show here that Clarithromycin impairs both the frequencies and number of interleukin (IL)-17 producing T helper (Th) 17 cells within the lungs of mice infected with a macrolide-resistant S. pneumoniae serotype 15A strain. Subsequently, the tissue-resident memory CD4+ T cell (Trm) response to a consecutive S. pneumoniae infection was impaired. The number of lung resident IL-17+ CD69+ Trm was diminished upon Clarithromycin treatment during reinfection. Mechanistically, Clarithromycin attenuated phosphorylation of the p90-S6-kinase as part of the ERK pathway in Th17 cells. Moreover, a strong increase in the mitochondrial-mediated maximal respiratory capacity was observed, while mitochondrial protein translation and mTOR sisgnaling were unimpaired. Therefore, treatment with macrolide antibiotics may favor the spread of antimicrobial-resistant pathogens not only by applying a selection pressure but also by decreasing the natural T cell immune response. Clinical administration of macrolide antibiotics as standard therapy procedure during initial hospitalization should be reconsidered accordingly and possibly be withheld until microbial resistance is determined. KEY MESSAGES: • Macrolide-resistant S. pneumoniae infection undergoes immunomodulation by Clarithromycin • Clarithromycin treatment hinders Th17 and tissue-resident memory responses • Macrolide antibiotics impair Th17 differentiation in vitro by ERK-pathway inhibition.
2021-03-30
2021-03-30
2021-02-17
Article
J Mol Med (Berl). 2021 Feb 17. doi: 10.1007/s00109-021-02039-5. Epub ahead of print.
33595670
10.1007/s00109-021-02039-5
http://hdl.handle.net/10033/622804
1432-1440
Journal of molecular medicine (Berlin, Germany)
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
Springer
oai:repository.helmholtz-hzi.de:10033/6228612021-05-08T01:43:01Zcom_10033_6839col_10033_621495
Conservation of the HBV RNA element epsilon in nackednaviruses reveals ancient origin of protein-primed reverse transcription.
Beck, Jürgen
Seitz, Stefan
Lauber, Chris
Nassal, Michael
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
HBV long-term evolution
HBV replication mechanism
initiation of reverse transcription
paleovirology
protein priming
Hepadnaviruses, with the human hepatitis B virus as prototype, are small, enveloped hepatotropic DNA viruses which replicate by reverse transcription of an RNA intermediate. Replication is initiated by a unique protein-priming mechanism whereby a hydroxy amino acid side chain of the terminal protein (TP) domain of the viral polymerase (P) is extended into a short DNA oligonucleotide, which subsequently serves as primer for first-strand synthesis. A key component in the priming of reverse transcription is the viral RNA element epsilon, which contains the replication origin and serves as a template for DNA primer synthesis. Here, we show that recently discovered non-enveloped fish viruses, termed nackednaviruses [C. Lauber et al., Cell Host Microbe 22, 387-399 (2017)], employ a fundamentally similar replication mechanism despite their huge phylogenetic distance and major differences in genome organization and viral lifestyle. In vitro cross-priming studies revealed that few strategic nucleotide substitutions in epsilon enable site-specific protein priming by heterologous P proteins, demonstrating that epsilon is functionally conserved since the two virus families diverged more than 400 Mya. In addition, other cis elements crucial for the hepadnavirus-typical replication of pregenomic RNA into relaxed circular double-stranded DNA were identified at conserved positions in the nackednavirus genomes. Hence, the replication mode of both hepadnaviruses and nackednaviruses was already established in their Paleozoic common ancestor, making it a truly ancient and evolutionary robust principle of genome replication that is more widespread than previously thought.
2021-05-07
2021-05-07
2021-03-30
Article
Proc Natl Acad Sci U S A. 2021 Mar 30;118(13):e2022373118. doi: 10.1073/pnas.2022373118.
33753499
10.1073/pnas.2022373118
http://hdl.handle.net/10033/622861
1091-6490
Proceedings of the National Academy of Sciences of the United States of America
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
Academy of Sciences
oai:repository.helmholtz-hzi.de:10033/6228632021-05-12T02:38:36Zcom_10033_6839col_10033_621495
NeutrobodyPlex-monitoring SARS-CoV-2 neutralizing immune responses using nanobodies.
Wagner, Teresa R
Ostertag, Elena
Kaiser, Philipp D
Gramlich, Marius
Ruetalo, Natalia
Junker, Daniel
Haering, Julia
Traenkle, Bjoern
Becker, Matthias
Dulovic, Alex
Schweizer, Helen
Nueske, Stefan
Scholz, Armin
Zeck, Anne
Schenke-Layland, Katja
Nelde, Annika
Strengert, Monika
Walz, Juliane S
Zocher, Georg
Stehle, Thilo
Schindler, Michael
Schneiderhan-Marra, Nicole
Rothbauer, Ulrich
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
SARS-CoV-2
immune response
nanobodies
neutralizing antibodies
serological assay
In light of the COVID-19 pandemic, there is an ongoing need for diagnostic tools to monitor the immune status of large patient cohorts and the effectiveness of vaccination campaigns. Here, we present 11 unique nanobodies (Nbs) specific for the SARS-CoV-2 spike receptor-binding domain (RBD), of which 8 Nbs potently inhibit the interaction of RBD with angiotensin-converting enzyme 2 (ACE2) as the major viral docking site. Following detailed epitope mapping and structural analysis, we select two inhibitory Nbs, one of which binds an epitope inside and one of which binds an epitope outside the RBD:ACE2 interface. Based on these, we generate a biparatopic nanobody (bipNb) with viral neutralization efficacy in the picomolar range. Using bipNb as a surrogate, we establish a competitive multiplex binding assay ("NeutrobodyPlex") for detailed analysis of the presence and performance of neutralizing RBD-binding antibodies in serum of convalescent or vaccinated patients. We demonstrate that NeutrobodyPlex enables high-throughput screening and detailed analysis of neutralizing immune responses in infected or vaccinated individuals, to monitor immune status or to guide vaccine design.
2021-05-11
2021-05-11
2021-04-27
Article
EMBO Rep. 2021 May 5;22(5):e52325. doi: 10.15252/embr.202052325. Epub 2021 Apr 27.
33904225
10.15252/embr.202052325
http://hdl.handle.net/10033/622863
1469-3178
EMBO reports
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
EMBO Press
oai:repository.helmholtz-hzi.de:10033/6228722021-05-18T01:59:41Zcom_10033_6839col_10033_621495
Direct conversion of porcine primary fibroblasts into hepatocyte-like cells.
Fráguas-Eggenschwiler, Mariane
Eggenschwiler, Reto
Söllner, Jenny-Helena
Cortnumme, Leon
Vondran, Florian W R
Cantz, Tobias
Ott, Michael
Niemann, Heiner
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
The pig is an important model organism for biomedical research, mainly due to its extensive genetic, physiological and anatomical similarities with humans. Until date, direct conversion of somatic cells into hepatocyte-like cells (iHeps) has only been achieved in rodents and human cells. Here, we employed lentiviral vectors to screen a panel of 12 hepatic transcription factors (TF) for their potential to convert porcine fibroblasts into hepatocyte-like cells. We demonstrate for the first time, hepatic conversion of porcine somatic cells by over-expression of CEBPα, FOXA1 and HNF4α2 (3TF-piHeps). Reprogrammed 3TF-piHeps display a hepatocyte-like morphology and show functional characteristics of hepatic cells, including albumin secretion, Dil-AcLDL uptake, storage of lipids and glycogen and activity of cytochrome P450 enzymes CYP1A2 and CYP2C33 (CYP2C9 in humans). Moreover, we show that markers of mature hepatocytes are highly expressed in 3TF-piHeps, while fibroblastic markers are reduced. We envision piHeps as useful cell sources for future studies on drug metabolism and toxicity as well as in vitro models for investigation of pig-to-human infectious diseases.
2021-05-17
2021-05-17
2021-04-29
Article
Sci Rep. 2021 Apr 29;11(1):9334. doi: 10.1038/s41598-021-88727-1.
33927320
10.1038/s41598-021-88727-1
http://hdl.handle.net/10033/622872
2045-2322
Scientific reports
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
Nature Research
oai:repository.helmholtz-hzi.de:10033/6229152021-07-02T01:42:46Zcom_10033_6839com_10033_620656col_10033_620657col_10033_621495
Structure-Activity Relationship and Mode-of-Action Studies Highlight 1-(4-Biphenylylmethyl)-1H-imidazole-Derived Small Molecules as Potent CYP121 Inhibitors.
Walter, Isabell
Adam, Sebastian
Gentilini, Maria Virginia
Kany, Andreas M
Brengel, Christian
Thomann, Andreas
Sparwasser, Tim
Köhnke, Jesko
Hartmann, Rolf W
HIPS, Helmholtz-Institut für Pharmazeutische Forschung Saarland, Universitätscampus E8.1 66123 Saarbrücken, Germany.; TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
CYP121
Mycobacterium tuberculosis
biological activity
complex structures
structure-activity relationships
CYP121 of Mycobacterium tuberculosis (Mtb) is an essential target for the development of novel potent drugs against tuberculosis (TB). Besides known antifungal azoles, further compounds of the azole class were recently identified as CYP121 inhibitors with antimycobacterial activity. Herein, we report the screening of a similarity-oriented library based on the former hit compound, the evaluation of affinity toward CYP121, and activity against M. bovis BCG. The results enabled a comprehensive SAR study, which was extended through the synthesis of promising compounds and led to the identification of favorable features for affinity and/or activity and hit compounds with 2.7-fold improved potency. Mode of action studies show that the hit compounds inhibit substrate conversion and highlighted CYP121 as the main antimycobacterial target of our compounds. Exemplified complex crystal structures of CYP121 with three inhibitors reveal a common binding site. Engaging in both hydrophobic interactions as well as hydrogen bonding to the sixth iron ligand, our compounds block a solvent channel leading to the active site heme. Additionally, we report the first CYP inhibitors that are able to reduce the intracellular replication of M. bovis BCG in macrophages, emphasizing their potential as future drug candidates against TB.
2021-07-01
2021-07-01
2021-05-19
Article
ChemMedChem. 2021 May 19. doi: 10.1002/cmdc.202100283. Epub ahead of print.
34010508
10.1002/cmdc.202100283
http://hdl.handle.net/10033/622915
1860-7187
ChemMedChem
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
Wiley-VCH
oai:repository.helmholtz-hzi.de:10033/6229642021-07-29T01:53:54Zcom_10033_620597com_10033_6839com_10033_621852col_10033_621853col_10033_620598col_10033_621495
Integration of metabolomics, genomics, and immune phenotypes reveals the causal roles of metabolites in disease.
Chu, Xiaojing
Jaeger, Martin
Beumer, Joep
Bakker, Olivier B
Aguirre-Gamboa, Raul
Oosting, Marije
Smeekens, Sanne P
Moorlag, Simone
Mourits, Vera P
Koeken, Valerie A C M
de Bree, Charlotte
Jansen, Trees
Mathews, Ian T
Dao, Khoi
Najhawan, Mahan
Watrous, Jeramie D
Joosten, Irma
Sharma, Sonia
Koenen, Hans J P M
Withoff, Sebo
Jonkers, Iris H
Netea-Maier, Romana T
Xavier, Ramnik J
Franke, Lude
Xu, Cheng-Jian
Joosten, Leo A B
Sanna, Serena
Jain, Mohit
Kumar, Vinod
Clevers, Hans
Wijmenga, Cisca
Netea, Mihai G
Li, Yang
CiiM, Zentrum für individualisierte Infektionsmedizin, Feodor-Lynen-Str.7, 30625 Hannover.; TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.; BRICS, Braunschweiger Zentrum für Systembiologie, Rebenring 56,38106 Braunschweig, Germany.
Genomics
Immune phenotypes
Integrative analysis
Metabolomics
Background: Recent studies highlight the role of metabolites in immune diseases, but it remains unknown how much of this effect is driven by genetic and non-genetic host factors.
Result: We systematically investigate circulating metabolites in a cohort of 500 healthy subjects (500FG) in whom immune function and activity are deeply measured and whose genetics are profiled. Our data reveal that several major metabolic pathways, including the alanine/glutamate pathway and the arachidonic acid pathway, have a strong impact on cytokine production in response to ex vivo stimulation. We also examine the genetic regulation of metabolites associated with immune phenotypes through genome-wide association analysis and identify 29 significant loci, including eight novel independent loci. Of these, one locus (rs174584-FADS2) associated with arachidonic acid metabolism is causally associated with Crohn's disease, suggesting it is a potential therapeutic target.
Conclusion: This study provides a comprehensive map of the integration between the blood metabolome and immune phenotypes, reveals novel genetic factors that regulate blood metabolite concentrations, and proposes an integrative approach for identifying new disease treatment targets.
2021-07-28
2021-07-28
2021-07-06
Article
Genome Biol. 2021 Jul 6;22(1):198. doi: 10.1186/s13059-021-02413-z.
34229738
10.1186/s13059-021-02413-z
http://hdl.handle.net/10033/622964
1474-760X
Genome biology
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
BMC
oai:repository.helmholtz-hzi.de:10033/6229932021-08-21T02:11:14Zcom_10033_6839col_10033_621495
Staphylococcus epidermidis Phages Transduce Antimicrobial Resistance Plasmids and Mobilize Chromosomal Islands.
Fišarová, Lenka
Botka, Tibor
Du, Xin
Mašlaňová, Ivana
Bárdy, Pavol
Pantůček, Roman
Benešík, Martin
Roudnický, Pavel
Winstel, Volker
Larsen, Jesper
Rosenstein, Ralf
Peschel, Andreas
Doškař, Jiří
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Staphylococcus epidermidis
antibiotic resistance
bacteriophages
horizontal gene transfer
pathogenicity islands
transduction
Staphylococcus epidermidis is a leading opportunistic pathogen causing nosocomial infections that is notable for its ability to form a biofilm and for its high rates of antibiotic resistance. It serves as a reservoir of multiple antimicrobial resistance genes that spread among the staphylococcal population by horizontal gene transfer such as transduction. While phage-mediated transduction is well studied in Staphylococcus aureus, S. epidermidis transducing phages have not been described in detail yet. Here, we report the characteristics of four phages, 27, 48, 456, and 459, previously used for S. epidermidis phage typing, and the newly isolated phage E72, from a clinical S. epidermidis strain. The phages, classified in the family Siphoviridae and genus Phietavirus, exhibited an S. epidermidis-specific host range, and together they infected 49% of the 35 strains tested. A whole-genome comparison revealed evolutionary relatedness to transducing S. aureus phietaviruses. In accordance with this, all the tested phages were capable of transduction with high frequencies up to 10-4 among S. epidermidis strains from different clonal complexes. Plasmids with sizes from 4 to 19 kb encoding resistance to streptomycin, tetracycline, and chloramphenicol were transferred. We provide here the first evidence of a phage-inducible chromosomal island transfer in S. epidermidis Similarly to S. aureus pathogenicity islands, the transfer was accompanied by phage capsid remodeling; however, the interfering protein encoded by the island was distinct. Our findings underline the role of S. epidermidis temperate phages in the evolution of S. epidermidis strains by horizontal gene transfer, which can also be utilized for S. epidermidis genetic studies.IMPORTANCE Multidrug-resistant strains of S. epidermidis emerge in both nosocomial and livestock environments as the most important pathogens among coagulase-negative staphylococcal species. The study of transduction by phages is essential to understanding how virulence and antimicrobial resistance genes spread in originally commensal bacterial populations. In this work, we provide a detailed description of transducing S. epidermidis phages. The high transduction frequencies of antimicrobial resistance plasmids and the first evidence of chromosomal island transfer emphasize the decisive role of S. epidermidis phages in attaining a higher pathogenic potential of host strains. To date, such importance has been attributed only to S. aureus phages, not to those of coagulase-negative staphylococci. This study also proved that the described transducing bacteriophages represent valuable genetic modification tools in S. epidermidis strains where other methods for gene transfer fail.
2021-08-20
2021-08-20
2021-05-12
2021-05-15
Article
mSphere. 2021 May 12;6(3):e00223-21. doi: 10.1128/mSphere.00223-21.
33980677
10.1128/mSphere.00223-21
http://hdl.handle.net/10033/622993
2379-5042
mSphere
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
American Society of Microbiology
oai:repository.helmholtz-hzi.de:10033/6230352021-09-17T03:40:30Zcom_10033_6839col_10033_621495
MicroRNA-125b-5p Regulates Hepatocyte Proliferation During the Termination Phase of Liver Regeneration.
Yang, Dakai
Dai, Zhen
Yang, Taihua
Balakrishnan, Asha
Yuan, Qinggong
Vondran, Florian W R
Manns, Michael P
Ott, Michael
Cantz, Tobias
Sharma, Amar Deep
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
The ability of the liver to regenerate and restore mass limits the increasing mortality rate due to life-threatening liver diseases. Successful liver regeneration is accomplished in multiple stages, of which the priming and proliferation phases are well studied. However, the regulatory pathways, specifically microRNA (miRNA)-mediated posttranscriptional regulation, which prevent uncontrolled proliferation and mediate the termination of liver regeneration, are not well understood. We identified differentially regulated miRNAs during the termination phase after 2/3 partial hepatectomy (PH) in mice, which is a well-established mouse model of liver regeneration. We further evaluated the function of differentially regulated miRNAs in primary mouse hepatocytes by using mimics and inhibitors and in vivo by using adeno-associated virus (AAV) serotype 8. A candidate miRNA target was identified by messenger RNA array in silico analyses and validated in primary mouse and human hepatocytes. Using miRNA profiling, we discovered miR-125b-5p as a novel regulator of hepatocyte proliferation in the late phase of liver regeneration. AAV-mediated miR-125b-5p delivery in mice enhanced the endogenous regenerative capacity and resulted in improved restoration of liver mass after 2/3 PH. Further, we found that ankyrin repeat and BTB/POZ domain containing protein 1 (Abtb1) is a direct target of miR-125b-5p in primary mouse and human hepatocytes and contributes to the pro-proliferative activity of miR-125b-5p by forkhead box G1 (FOXG1) and the cyclin-dependent kinase inhibitor 1A (p21) pathway. Conclusion: miR-125b-5p has an important role in regulating hepatocyte proliferation in the termination phase of liver regeneration and may serve as a potential therapeutic target in various liver diseases that often exhibit deregulated hepatocyte proliferation.
2021-09-16
2021-09-16
2020-09-15
Article
Hepatol Commun. 2020 Sep 15;4(12):1851-1863. doi: 10.1002/hep4.1597.
33305155
10.1002/hep4.1597
http://hdl.handle.net/10033/623035
2471-254X
Hepatology communications
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
Wiley
oai:repository.helmholtz-hzi.de:10033/6230422021-09-27T11:01:38Zcom_10033_6839col_10033_621495
Transient Depletion of Foxp3 Regulatory T Cells Selectively Promotes Aggressive β Cell Autoimmunity in Genetically Susceptible DEREG Mice.
Watts, Deepika
Janßen, Marthe
Jaykar, Mangesh
Palmucci, Francesco
Weigelt, Marc
Petzold, Cathleen
Hommel, Angela
Sparwasser, Tim
Bonifacio, Ezio
Kretschmer, Karsten
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Foxp3
Treg cells
cell ablation
immune regulation
type 1 diabetes
Type 1 diabetes (T1D) represents a hallmark of the fatal multiorgan autoimmune syndrome affecting humans with abrogated Foxp3+ regulatory T (Treg) cell function due to Foxp3 gene mutations, but whether the loss of Foxp3+ Treg cell activity is indeed sufficient to promote β cell autoimmunity requires further scrutiny. As opposed to human Treg cell deficiency, β cell autoimmunity has not been observed in non-autoimmune-prone mice with constitutive Foxp3 deficiency or after diphtheria toxin receptor (DTR)-mediated ablation of Foxp3+ Treg cells. In the spontaneous nonobese diabetic (NOD) mouse model of T1D, constitutive Foxp3 deficiency did not result in invasive insulitis and hyperglycemia, and previous studies on Foxp3+ Treg cell ablation focused on Foxp3DTR NOD mice, in which expression of a transgenic BDC2.5 T cell receptor (TCR) restricted the CD4+ TCR repertoire to a single diabetogenic specificity. Here we revisited the effect of acute Foxp3+ Treg cell ablation on β cell autoimmunity in NOD mice in the context of a polyclonal TCR repertoire. For this, we took advantage of the well-established DTR/GFP transgene of DEREG mice, which allows for specific ablation of Foxp3+ Treg cells without promoting catastrophic autoimmune diseases. We show that the transient loss of Foxp3+ Treg cells in prediabetic NOD.DEREG mice is sufficient to precipitate severe insulitis and persistent hyperglycemia within 5 days after DT administration. Importantly, DT-treated NOD.DEREG mice preserved many clinical features of spontaneous diabetes progression in the NOD model, including a prominent role of diabetogenic CD8+ T cells in terminal β cell destruction. Despite the severity of destructive β cell autoimmunity, anti-CD3 mAb therapy of DT-treated mice interfered with the progression to overt diabetes, indicating that the novel NOD.DEREG model can be exploited for preclinical studies on T1D under experimental conditions of synchronized, advanced β cell autoimmunity. Overall, our studies highlight the continuous requirement of Foxp3+ Treg cell activity for the control of genetically pre-installed autoimmune diabetes.
2021-09-22
2021-09-22
2021-08-10
Article
Front Immunol. 2021 Aug 10;12:720133. doi: 10.3389/fimmu.2021.720133.
34447385
10.3389/fimmu.2021.720133
http://hdl.handle.net/10033/623042
1664-3224
Frontiers in immunology
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
Frontiers
oai:repository.helmholtz-hzi.de:10033/6230622021-10-06T01:56:53Zcom_10033_6839col_10033_621495
Therapeutic HNF4A mRNA attenuates liver fibrosis in a preclinical model.
Yang, Taihua
Poenisch, Marion
Khanal, Rajendra
Hu, Qingluan
Dai, Zhen
Li, Ruomeng
Song, Guangqi
Yuan, Qinggong
Yao, Qunyan
Shen, Xizhong
Taubert, Richard
Engel, Bastian
Jaeckel, Elmar
Vogel, Arndt
Falk, Christine S
Schambach, Axel
Gerovska, Daniela
Araúzo-Bravo, Marcos J
Vondran, Florian W R
Cantz, Tobias
Horscroft, Nigel
Balakrishnan, Asha
Chevessier, Frédéric
Ott, Michael
Sharma, Amar Deep
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Transcription factors
mRNA therapeutics
protein replacement and cirrhosis
Background and aims: Messenger RNA (mRNA)-based therapeutics are rapidly progressing to the clinic and hold tremendous potential for benefiting millions of people worldwide. Therapeutic targeting of injuries that require transient restoration of proteins by mRNA delivery is an attractive aspect, however until recently, it has remained poorly explored. In this study, we examined for the first time, the therapeutic utility of mRNA delivery in liver fibrosis and cirrhosis, which contributes to millions of deaths, annually. Here, we aimed to demonstrate therapeutic efficacy of the human transcription factor hepatocyte nuclear factor alpha (HNF4A) encoding mRNA in chronically injured murine liver leading to fibrosis and cirrhosis.
Methods: We investigated restoration of hepatocyte functions by HNF4A mRNA transfection in vitro, and analyzed the attenuation of liver fibrosis and cirrhosis in multiple mouse models, by delivering hepatocyte-targeted biodegradable lipid nanoparticles (LNP) encapsulating HNF4A mRNA. To identify potential mechanisms, we performed microarray-based gene expression profiling, single cell RNA sequencing, and chromatin immunoprecipitation. We used primary liver cells and human liver buds for additional functional validation.
Results: Expression of HNF4A encoding mRNA led to restoration of metabolic activity of fibrotic primary murine and human hepatocytes in vitro. Repeated in vivo delivery of HNF4A mRNA encapsulated-LNP induced a robust inhibition of fibrogenesis in four independent mouse models of hepatotoxin- and cholestasis-induced liver fibrosis. Mechanistically, we discovered that paraoxonase 1 is a direct target of HNF4A and it contributes to HNF4A-mediated attenuation of liver fibrosis via modulation of liver macrophages and hepatic stellate cells.
Conclusion: Collectively, our findings provide the first direct preclinical evidence of the applicability of HNF4A mRNA therapeutics for the treatment of fibrosis in the liver.
Lay summary: Liver fibrosis and cirrhosis remain unmet medical needs and contribute to high mortality, worldwide. Herein, we take advantage of a promising, emerging mRNA therapy approach to treat liver fibrosis and cirrhosis. We demonstrate that restoration of a key gene, HNF4A, via mRNA encapsulated in lipid nanoparticles decreased injury in multiple mouse models of fibrosis and cirrhosis. Our study provides the proof-of-concept that mRNA therapy would be a promising strategy for reversing liver fibrosis and cirrhosis.
2021-10-05
2021-10-05
2021-08-25
Article
J Hepatol. 2021 Aug 25:S0168-8278(21)02006-7. doi: 10.1016/j.jhep.2021.08.011. Epub ahead of print.
34453962
10.1016/j.jhep.2021.08.011
http://hdl.handle.net/10033/623062
1600-0641
Journal of hepatology
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
Elsevier
oai:repository.helmholtz-hzi.de:10033/6230712021-10-13T03:22:02Zcom_10033_6839col_10033_621495
Therapeutic HNF4A mRNA attenuates liver fibrosis in a preclinical model.
Yang, Taihua
Poenisch, Marion
Khanal, Rajendra
Hu, Qingluan
Dai, Zhen
Li, Ruomeng
Song, Guangqi
Yuan, Qinggong
Yao, Qunyan
Shen, Xizhong
Taubert, Richard
Engel, Bastian
Jaeckel, Elmar
Vogel, Arndt
Falk, Christine S
Schambach, Axel
Gerovska, Daniela
Araúzo-Bravo, Marcos J
Vondran, Florian W R
Cantz, Tobias
Horscroft, Nigel
Balakrishnan, Asha
Chevessier, Frédéric
Ott, Michael
Sharma, Amar Deep
TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.
Transcription factors
mRNA therapeutics
protein replacement and cirrhosis
Background & aims: Therapeutic targeting of injuries that require transient restoration of proteins by mRNA delivery is an attractive approach that, until recently, has remained poorly explored. In this study, we examined the therapeutic utility of mRNA delivery for liver fibrosis and cirrhosis. Specifically, we aimed to demonstrate the therapeutic efficacy of human hepatocyte nuclear factor alpha (HNF4A) mRNA in mouse models of fibrosis and cirrhosis.
Methods: We investigated restoration of hepatocyte functions by HNF4A mRNA transfection in vitro, and analyzed the attenuation of liver fibrosis and cirrhosis in multiple mouse models, by delivering hepatocyte-targeted biodegradable lipid nanoparticles (LNPs) encapsulating HNF4A mRNA. To identify potential mechanisms of action, we performed microarray-based gene expression profiling, single-cell RNA sequencing, and chromatin immunoprecipitation. We used primary liver cells and human liver buds for additional functional validation.
Results: Expression of HNF4A mRNA led to restoration of the metabolic activity of fibrotic primary murine and human hepatocytes in vitro. Repeated in vivo delivery of LNP-encapsulated HNF4A mRNA induced a robust inhibition of fibrogenesis in 4 independent mouse models of hepatotoxin- and cholestasis-induced liver fibrosis. Mechanistically, we discovered that paraoxonase 1 is a direct target of HNF4A and it contributes to HNF4A-mediated attenuation of liver fibrosis via modulation of liver macrophages and hepatic stellate cells.
Conclusion: Collectively, our findings provide the first direct preclinical evidence of the applicability of HNF4A mRNA therapeutics for the treatment of fibrosis in the liver.
Lay summary: Liver fibrosis and cirrhosis remain unmet medical needs and contribute to high mortality worldwide. Herein, we take advantage of a promising therapeutic approach to treat liver fibrosis and cirrhosis. We demonstrate that restoration of a key gene, HNF4A, via mRNA encapsulated in lipid nanoparticles decreased injury in multiple mouse models of fibrosis and cirrhosis. Our study provides proof-of-concept that mRNA therapy is a promising strategy for reversing liver fibrosis and cirrhosis.
2021-10-12
2021-10-12
2021-08-25
Article
J Hepatol. 2021 Aug 25:S0168-8278(21)02006-7. doi: 10.1016/j.jhep.2021.08.011. Epub ahead of print.
34453962
10.1016/j.jhep.2021.08.011
http://hdl.handle.net/10033/623071
1600-0641
Journal of hepatology
en
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Elsevier
oai:repository.helmholtz-hzi.de:10033/6230762021-10-21T03:21:07Zcom_10033_6839com_10033_621852col_10033_621853col_10033_621495
Evolution of cytokine production capacity in ancient and modern European populations.
Domínguez-Andrés, Jorge
Kuijpers, Yunus
Bakker, Olivier B
Jaeger, Martin
Xu, Cheng-Jian
Van der Meer, Jos Wm
Jakobsson, Mattias
Bertranpetit, Jaume
Joosten, Leo Ab
Li, Yang
Netea, Mihai G
CSSB, Centre for Structural Systembiologie, Notkestr.85, 22607 Hamburg. Germany.
adaptation
disease
evolution
genetics
genomics
human
immune system
immunology
inflammation
neolithic
As our ancestors migrated throughout different continents, natural selection increased the presence of alleles advantageous in the new environments. Heritable variations that alter the susceptibility to diseases vary with the historical period, the virulence of the infections, and their geographical spread. In this study we built polygenic scores for heritable traits that influence the genetic adaptation in the production of cytokines and immune-mediated disorders, including infectious, inflammatory, and autoimmune diseases, and applied them to the genomes of several ancient European populations. We observed that the advent of the Neolithic was a turning point for immune-mediated traits in Europeans, favoring those alleles linked with the development of tolerance against intracellular pathogens and promoting inflammatory responses against extracellular microbes. These evolutionary patterns are also associated with an increased presence of traits related to inflammatory and auto-immune diseases.
2021-10-20
2021-10-20
2021-09-07
Article
Elife. 2021 Sep 7;10:e64971. doi: 10.7554/eLife.64971.
34488939
10.7554/eLife.64971
http://hdl.handle.net/10033/623076
2050-084X
eLife
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
eLife Sciences Publications
oai:repository.helmholtz-hzi.de:10033/6232052022-06-14T01:56:56Zcom_10033_6839col_10033_621495
Guidance for design and endpoints of clinical trials in chronic hepatitis B - Report from the 2019 EASL-AASLD HBV Treatment Endpoints Conference.
Cornberg, Markus
Lok, Anna Suk-Fong
Terrault, Norah A
Zoulim, Fabien
Antiviral therapy
CpAM
Hepatitis B surface antigen
Hepatitis D
Immunomodulatory therapy
Nucleos(t)ide analogues
2022-06-13
2022-06-13
2020-03
2019-07-10
Article
Other
31730789
10.1016/j.jhep.2019.11.003
http://hdl.handle.net/10033/623205
1600-0641
Journal of hepatology
en
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International