TP11: Effects of constitutive cytokine signalling on tissue homeostasis and liver cell-intrinsic responsiveness to hepatitis virus infection

We have previously discovered a major impact of the cytokine milieu onto the sensitivity of tissue cells towards virus infection. We will now study the interplay of immune cells as major producers of cytokines and parenchymal cells. We hypothesize that inter-individual differences in immune cell activity might decisively shape the antiviral responsiveness in the liver, and contribute to the clinical outcome of HCV infection (acute self-resolving vs. persisting infection). We will further investigate the molecular mechanisms behind hepatocyte priming, and analyse the reciprocal interaction between parenchymal cells and immune cells.

Dr. Claudia Beisel

Dr. Marco Binder

Urban C*, Welsch H*, Heine K, Wüst S, Haas DA, Dächert C, Pandey A, Pichlmair A*, Binder M.* (2020) Persistent Innate Immune Stimulation Results in IRF3-Mediated but Caspase-Independent Cytostasis. Viruses. 12, 635, doi: 10.3390/v12060635.

Burkart SS, Schweinoch D, Frankish J, Sparn C, Wüst S, Urban C, Merlo M, Magalhães VG, Piras A, Pichlmair A, Willemsen J, Kaderali L, Binder M. (2023) High-resolution kinetic characterization of the RIG-I-signaling pathway and the antiviral response. Life Sci Alliance. 6., doi: 10.26508/lsa.202302059.

Wüst S, Schad P, Burkart SS, Binder M. (2021) Comparative Analysis of Six IRF Family Members in Alveolar Epithelial Cell-Intrinsic Antiviral Responses. Cells. 10, 2600, doi: 10.3390/cells10102600.

Magalhães VG, Lukassen S, Drechsler M, Loske J, Burkart SS, Wüst S, Jacobsen E-M, Röhmel J, Mall MA, Debatin K-M, Eils R, Autenrieth S, Janda A, Lehmann I, Binder M. (2023) Enhanced Airway Epithelial Response to SARS-CoV-2 Infection in Children is Critically Tuned by the Cross-Talk Between Immune and Epithelial Cells. EMBO Rep. e57912 doi: 10.15252/embr.202357912.

Laudenbach BT, Krey K, Emslander Q, Andersen LL, Reim A, Scaturro P, Mundigl S, Dächert C, Manske K, Moser M, Ludwig J, Wohlleber D, Kröger A, Binder M, Pichlmair A. (2021) NUDT2 initiates viral RNA degradation by removal of 5′-phosphates. Nat Commun. 12, 6918, doi:10.1038/s41467-021-27239-y.

Kok F, Rosenblatt M, Teusel M, Nizharadze T, Magalhães VG, Dächert C, Maiwald T, Vlasov A, Wäsch M, Tyufekchieva S, Hoffmann K, Damm G, Seehofer D, Boettler T, Binder M, Timmer J, Schilling M, Klingmüller U. (2020) Disentangling molecular mechanisms regulating sensitization of interferon alpha signal transduction. Mol Syst Biol. 16, e8955, doi: 10.15252/msb.20198955.

Loske J, Röhmel J, Lukassen S, Stricker S, Magalhães VG, Liebig J, Chua RL, Thürmann L, Messingschlager M, Seegebarth A, Timmermann B, Klages S, Ralser M, Sawitzki B, Sander LE, Corman VM, Conrad C, Laudi S, Binder M*, Trump S*, Eils R*, Mall MA*, Lehmann I.* (2022) Pre-activated antiviral innate immunity in the upper airways controls early SARS-CoV-2 infection in children. Nat Biotechnol. 40, 319–324, doi:10.1038/s41587-021-01037-9.

Cords L, Woost R, Kummer S, Brehm TT, Kluge S, Schmiedel S, Jordan S, Lohse AW, Altfeld M, Addo MM, Schulze zur Wiesch J, Beisel C. (2023) Frequency of IRF5+ dendritic cells is associated with the TLR7-induced inflammatory cytokine response in SARS-CoV-2 infection. Cytokine. 162, 156109–156109, doi: 10.1016/j.cyto.2022.156109.

Beisel C, Jordan-Paiz A, Köllmann S, Ahrenstorf AE, Padoan B, Barkhausen T, Addo MM, Altfeld, M. (2023) Sex differences in the percentage of IRF5 positive B cells are associated with higher production of TNF-α in women in response to TLR9 in humans. Biol Sex Differ. 14,11, doi:10.1186/s13293-023-00495-x.

Beisel C, Ziegler S, Zapater GM, Chapel A, Griesbeck M, Hildebrandt H, Lohse AW, Altfeld M. (2017) TLR7-mediated activation of XBP1 correlates with the IFNα production in humans. Cytokine. 94, 55–58, doi:10.1016/j.cyto.2017.04.006.

Project in detail +

In this funding period we will expand on our previous work by widening our focus to immune cells as important producers of cytokines. We will test their capacity to prime hepatocytes to increase tissue responsiveness towards hepatitis virus infection. Analogously to what we observed in the respiratory epithelium with SarsCoV2, inter-individual differences in cytokine production (qualitative and quantitative) by innate immune cells at baseline might decisively tune hepatocyte-intrinsic antiviral sensing. In particular for HCV, which is spontaneously cleared in only 20% of cases, we hypothesize that such inter-individual differences significantly contribute to the differential capacity of patients to clear the infection. We will test this hypothesis using peripheral blood mononuclear cells (PBMCs) of individuals with a defined HCV history. For this, the new Co-PI Beisel joined TP11, who is a clinician scientist specializing in viral hepatitis with access to existing patient samples as well as to newly presenting patients. We also collaborate with Thimme, who has access to further cohorts, e.g. through the FREEZE biobank in Freiburg. We will compare patients who spontaneously resolved HCV infection in the past versus patients who failed to control the virus and required DAA-treatment to clear it. Immune cells of the two groups will be functionally characterized by assessing cytokine profiles at baseline as well as upon stimulation with TLR ligands, and their ability to prime hepatocytes and inhibit HCV infection in vitro.
We will further investigate the general molecular mechanism of cytokine priming of parenchymal cells at a gene regulatory as well as functional level. Gene regulation will be studied in close collaboration with Z03, with whom we recently established single-cell multiomics (RNASeq + ATACSeq) to assess transcriptional and epigenetic changes induced by cytokine priming. Functionally, we will assess the effect of priming with individual cytokines or PBMC-derived authentic combinations of cytokines on the strength of the hepatocyte-intrinsic antiviral response and its impact on HCV, HDV and HEV infection (with Urban/Nkongolo, Bartenschlager and Dao Thi). Overcoming the limitations of monotypic cell-cultures, we will establish HLA-matched hepatocyte–PBMC co-cultures (with Dao Thi), and re evaluate induction of antiviral responses to HCV, HDV and HEV infection in such more authentic “tissue-like” conditions. In preliminary experiments we found cytokines secreted by infected tissue cells in turn influence co-cultured immune cells. Therefore, we hypothesize that a bi-directional parenchymal–immune cell communication plays a crucial and so-far underestimated role in determining innate antiviral responses. We will analyse the impact of hepatocyte-intrinsic antiviral signaling on the surrounding immune cells by single cell approaches incl. high-dimensional cytometry (with Z02), and scRNASeq (with Z03). In this context, an additional focus will lie on the effects of virus-encoded immune antagonists, including possible effects of secreted viral factors such as the glycosylated form of HEV ORF2 (with Dao Thi; collaboration already started) or the enigmatic HBV eAg onto immune cells (with Urban/Nkongolo, Protzer/Wettengel, Schütz/Seitz).