TP04: Regulation of virus-specific CD4 T helper cell responses in viral hepatitis

CD4 T helper cells are central players in the regulation of the antiviral immune response. They are particularly important for the maintenance of cytotoxic CD8 T cell responses and the emergence of antibody-producing B cells. Distinct subpopulations of the CD4 T helper compartment are involved in these functions. This project aims to define the composition of the CD4 T helper compartment in the setting of acute and chronic viral hepatitis. These observations could provide a blueprint of successful CD4 T cell responses that could inform vaccine design.

Prof (Apl) Dr. Tobias Böttler

Csernalabics B, Marinescu MS, Maurer L, Kelsch L, Werner J, Baumann K, Zoldan K, Panning M, Reuken P, Bruns T, Bengsch B, Neumann-Haefelin C, Hofmann M, Thimme R, Dao Thi VL, Boettler T. Efficient formation and maintenance of humoral and CD4 T cell immunity targeting the viral capsid in acute-resolving hepatitis E infection. J Hepatol. 2023 Dec 26:S0168-8278(23)05370-9. doi: 10.1016/j.jhep.2023.12.016. (TP01, TP02, TP04, TP20, TP21, TP22)

Winkler F, Hipp AV, Ramirez C, Martin B, Villa M, Neuwirt E, Gorka O, Aerssens J, Johansson SE, Rana N, Llewellyn-Lacey S, Price DA, Panning M, Gross O, Pearce EL, Herrmann CM, Schumann K, Hannibal L, Neumann-Haefelin C, Boettler T, Knolle P, Hofmann M, Wohlleber D, Thimme R, Bengsch B. Enolase represents a metabolic checkpoint controlling the differential exhaustion programs of hepatitis virus-specific CD8+ T cells. Gut accepted (TP01, TP02, TP04, TP05, TP13, TP20, TP21, Z03)

Luxenburger H, Reeg D, Lang-Meli J, Reinscheid M, Eisner M, Bettinger D, Oberhardt V, Alizei ES, Wild K,Graeser A, Karl V, Sagar, Emmerich F, Klein F, Panning M, Huzly D, Bengsch B, Boettler T, Elling R, Thimme R, Hofmann M, Neumann-Haefelin C. (2023) Boosting compromised SARS-CoV-2-specific immunity with mRNA vaccination in liver transplant recipients. J Hepatol. 78(5):1017-1027. doi: 10.1016/j.jhep.2023.02.007.

Reinscheid M, Luxenburger H, Karl V, Graeser A, Giese S, Ciminski K, Reeg DB, Oberhardt V, Roehlen N, Lang-Meli J, Heim K, Gross N, Baum C, Rieg S, Speer C, Emmerich F, Breisinger S, Steinmann D, Bengsch B, Boettler T, Kochs G, Schwemmle M, Thimme R, Neumann-Haefelin C, Hofmann M. (2022) COVID-19 mRNA booster vaccine induces transient CD8+ T effector cell responses while conserving the memory pool for subsequent reactivation. Nat Commun. 13(1):4631. doi: 10.1038/s41467-022-32324-x.

Barsch M, Salié H, Schlaak AE, Zhang Z, Hess M, Mayer LS, Tauber C, Otto-Mora P, Ohtani T, Nilsson T, Wischer L, Winkler F, Manne S, Rech A, Schmitt-Graeff A, Bronsert P, Hofmann M, Neumann-Haefelin C, Boettler T, Fichtner-Feigl S, van Boemmel F, Berg T, Rimassa L, Di Tommaso L, Saeed A, D'Alessio A, Pinato DJ, Bettinger D, Binder H, Wherry JE, Schultheiss M, Thimme R, Bengsch B. (2022) T-cell exhaustion and residency dynamics inform clinical outcomes in hepatocellular carcinoma. J Hepatol. 77(2):397-409. doi: 10.1016/j.jhep.2022.02.032.

Kemming J, Gundlach S, Panning M, Huzly D, Huang J, Lütgehetmann M, Pischke S, Schulze zur Wiesch J, Emmerich F, Llewellyn-Lacey S, Price DA, Tanriver Y, Warnatz K, Boettler T, Thimme R, Hofmann M, Fischer N, Neumann-Haefelin C. (2022) Mechanisms of CD8+ T-cell failure in chronic hepatitis E virus infection. J Hepatol. 77(4):978-990. https://doi.org/10.1016/j.jhep.2022.05.019.

Lang-Meli J, Luxenburger H, Wild K, Karl V, Oberhardt V, Salimi Alizei E, Graeser A, Reinscheid M, Roehlen N, Reeg DB, Giese S, Ciminski K, Götz V, August D, Rieg S, Waller CF, Wengenmayer T, Staudacher D, Huzly D, Bengsch B, Kochs G, Schwemmle M, Emmerich F, Boettler T, Thimme R, Hofmann M, Neumann-Haefelin C. (2022) SARS-CoV-2-specific T-cell epitope repertoire in convalescent and mRNA-vaccinated individuals. Nat Microbiol. 7(5):675-679. doi: 10.1038/s41564-022-01106-y.

Boettler T, Csernalabics B, Salié H, Luxenburger H, Wischer L, Alizei ES , Zoldan K, Krimmel L, Bronsert P, Schwabenland M, Prinz M, Mogler C, Neumann-Haefelin C, Thimme R, Hofmann M, Bengsch B. (2022) SARS-CoV-2 vaccination can elicit a CD8 T-cell dominant hepatitis. J Hepatol. 77(3):653-659 https://doi.org/10.1016/j.jhep.2022.03.040.

Boettler T , Gill U S, Allweiss L, Pollicino T, Tavis J E, Zoulim F. (2022) Assessing immunological and virological responses in the liver: Implications for the cure of chronic hepatitis B virus infection. JHEP Rep. Volume 4(6), 100480, ISSN 2589-5559. https://doi.org/10.1016/j.jhepr.2022.100480.

Oberhardt V, Hofmann M, Thimme R, Neumann-Haefelin C. (2022) Adaptive immune responses, immune escape and immune-mediated pathogenesis during HDV infection. Viruses. 20;14(2):198. doi: 10.3390/v14020198.

Wild K, Smits M, Killmer S, Strohmeier S, Neumann-Haefelin C, Bengsch B, Krammer F, Schwemmle M, Hofmann M, Thimme R, Zoldan K, Boettler T. (2021) Pre-existing immunity and vaccine history determine hemagglutinin-specific CD4 T cell and IgG response following seasonal influenza vaccination. Nat Commun. 18;12(1):6720. doi: 10.1038/s41467-021-27064-3.

Zoldan K, Ehrlich S, Killmer S, Wild K, Smits M, Russ M, Globig AM, Hofmann M, Thimme R, Boettler T. (2021) Th1-biased Hepatitis C virus-specific follicular T helper-like cells effectively support B cells after antiviral therapy. Front Immunol. 30;12:742061. doi: 10.3389/fimmu.2021.742061.

Schulien I, Kemming J, Oberhardt V, Wild K, Seidel LM, Killmer S, Sagar, Daul F, Salvat Lago M, Decker A, Luxenburger H, Binder B, Bettinger D, Sogukpinar O, Rieg S, Panning M, Huzly D, Schwemmle M, Kochs G, Waller CF, Nieters A, Duerschmied D, Emmerich F, Mei HE, Schulz AR, Llewellyn-Lacey S, Price DA, Boettler T, Bengsch B, Thimme R, Hofmann M, Neumann-Haefelin C. (2021) Characterization of pre-existing and induced SARS-CoV-2-specific CD8+ T cells. Nat Med. 27(1):78-85. doi: 10.1038/s41591-020-01143-2.

Oberhardt V, Luxenburger H, Kemming J, Schulien I, Ciminski K, Giese S, Csernalabics B, Lang-Meli J, Janowska I, Staniek J, Wild K, Basho K, Marinescu MS, Fuchs J, Topfstedt F, Janda A, Sogukpinar O, Hilger H, Stete K, Emmerich F, Bengsch B, Waller CF, Rieg S, Sagar, Boettler T, Zoldan K, Kochs G, Schwemmle M, Rizzi M, Thimme R, Neumann-Haefelin C, Hofmann M. (2021) Rapid and stable mobilization of CD8+ T cells by SARS-CoV-2 mRNA vaccine. Nature. 597(7875):268-273. doi: 10.1038/s41586-021-03841-4.

Schwabenland M, Salié H, Tanevski J, Killmer S, Lago MS, Schlaak AE, Mayer L, Matschke J, Püschel K, Fitzek A, Ondruschka B, Mei HE, Boettler T, Neumann-Haefelin C, Hofmann M, Breithaupt A, Genc N, Stadelmann C, Saez-Rodriguez J, Bronsert P, Knobeloch KP, Blank T, Thimme R, Glatzel M, Prinz M, Bengsch B. (2021) Deep spatial profiling of human COVID-19 brains reveals neuroinflammation with distinct microanatomical microglia-T cell interactions. Immunity. 13;54(7):1594-1610.e11. doi: 10.1016/j.immuni.2021.06.002.

Dudek M, Pfister D, Donakonda S, ....Thimme R, Böttler T, Tacke F, Dufour J-F, Haller D, Murray PJ, Heeren R, Zehn D, Böttcher JP, Heikenwälder M, Knolle PA. (2021) Auto-aggressive CXCR6+ CD8 T cells cause liver immune pathology in NASH. Nature. 592, 444–449. doi: 10.1038/s41586-021-03568-2.

Basho K, Zoldan K, Schultheiss M, Bettinger D, Globig A-M, Bengsch B, Neumann-Haefelin C, Klocperk A, Warnatz K, Hofmann M, Thimme R, Boettler T. (2021) IL-2 contributes to cirrhosis-associated immune dysfunction by impairing follicular T helper cells in advanced cirrhosis. J Hepatol. 74(3):649-660. doi: 10.1016/j.jhep.2020.10.012.

Hensel N, Gu Z, Sagar, Wieland D, Jechow K, Kemming J, Llewellyn-Lacey S, Gostick E, Sogukpinar O, Emmerich F, Price DA, Bengsch B, Boettler T, Neumann-Haefelin C, Eils R, Conrad C, Bartenschlager R, Grün D, Ishaque N, Thimme R, Hofmann M. (2021) Memory-like HCV-specific CD8+ T cells retain a molecular scar after cure of chronic HCV infection. Nat Immunol. 22(2):229-239. doi: 10.1038/s41590-020-00817-w.

Heim K, Binder B, Sagar, Wieland D, Hensel N, Llewellyn-Lacey S, Gostick E, Price DA, Emmerich F, Vingerhoet H, Kraft ARM, Cornberg M, Boettler T, Neumann-Haefelin C, Zehn D, Bengsch B, Hofmann M, Thimme R. (2020) TOX defines the degree of CD8+ T cell dysfunction in distinct phases of chronic HBV infection. Gut. 70(8): 1550–1560. doi: 10.1136/gutjnl-2020-322404

Basho K, Zoldan K, Schultheiss M, Bettinger D, Globig AM, Bengsch B, Neumann-Haefelin C, Klocperk A, Warnatz K, Hofmann M, Thimme R, Boettler T. (2020) IL-2 contributes to cirrhosis-associated immune dysfunction by impairing follicular T helper cells in advanced cirrhosis. J Hepatol. 24:S0168-8278(20)33692-8. doi: 10.1016/j.jhep.2020.10.012.

Kok F, Rosenblatt M, Teusel M, Nizharadze T, Gonçalves Magalhães V, 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(7):e8955. doi: 10.15252/msb.20198955.

Smits M, Zoldan K, Ishaque N, Gu Z, Jechow K, Wieland D, Conrad C, Eils R, Fauvelle C, Baumert TF, Emmerich F, Bengsch B, Neumann-Haefelin C, Hofmann M, Thimme R, Boettler T. 2020. Follicular T helper cells shape the HCV-specific CD4 T cell repertoire after viral elimination. J Clin Invest. 130(2):998-1009. (TP01, TP01A, TP02, TP04, Z02)

Smits M, Zoldan K, Wieland D, Ishaque N, Gu Z, Jechow K, Conrad C, Eils R, Fauvelle C, Baumert TF, Emmerich F, Bengsch B, Neumann-Haefelin C, Hofmann M, Thimme R, Boettler T. 2019. Follicular T helper cells shape HCV-specific CD4 T cell repertoire after viral elimination. J Clin Invest. 2019 Nov 7. pii: 129642. doi: 10.1172/JCI129642. [Epub ahead of print]

Ackermann C, Smits M, Woost R, Eberhard JM, Peine S, Kummer S, Marget M, Kuntzen T, Kwok WW, Lohse AW, Jacobs T, Boettler T, Schulze Zur Wiesch J. 2019. HCV-specific CD4+ T cells of patients with acute and chronic HCV infection display high expression of TIGIT and other co-inhibitory mole-cules. Sci Rep. 9(1):10624.

Jacobi FJ, Wild K, Smits M, Zoldan K, Csernalabics B, Flecken T, Lang J, Ehrenmann P, Emmerich F, Hofmann M, Thimme R, Neumann-Haefelin C, Boettler T. 2019. OX40 stimulation and PD-L1 blockade synergistically augment HBV-specific CD4 T cells in patients with HBeAg-negative infection. J Hepatol. 70(6):1103-1113.

Schuch A, Zecher BF, Müller PA, Correia MP, Daul F, Rennert C, Tauber C, Schlitt K, Boettler T, Neumann-Haefelin C, Hengel H, Pircher H, Cerwenka A, Thimme R, Hofmann M. 2019. NK-cell responses are biased towards CD16-mediated effector functions in chronic hepatitis B virus infection. J Hepatol. 70(3):351-360.

Luxenburger H, Graß F, Baermann J, Boettler T, Marget M, Emmerich F, Panning M, Thimme R, Nitsch-ke K, Neumann-Haefelin C. 2018. Differential virus-specific CD8(+)T-cell epitope repertoire in hepatitis C virus genotype 1 versus 4. J Viral Hepat. 25(7):779-790.

Raziorrouh B, Sacher K, Tawar RG, Emmerich F, Neumann-Haefelin C, Baumert TF, Thimme R, Boettler T. 2016. Virus-Specific CD4+ T Cells Have Functional and Phenotypic Characteristics of Follicular T-Helper Cells in Patients With Acute and Chronic HCV Infections. Gastroenterology. 150(3):696-706

Boettler T, Choi YS, Salek-Ardakani S, Cheng Y, Moeckel F, Croft M, Crotty S, von Herrath M. 2013. Exogenous OX40 stimulation during lymphocytic choriomeningitis virus infection impairs follicular Th cell differentiation and diverts CD4 T cells into the effector lineage by upregulating Blimp-1. J Immunol. 191(19):5026-35.

Boettler T, Moeckel F, Cheng Y, Heeg M, Salek-Ardakani S, Crotty S, Croft M, von Herrath MG. 2012. OX40 facilitates control of a persistent virus infection. PLoS Pathog. 8:e1002913.

Boettler T, Panther E, Bengsch B, Nazarova N, Spangenberg HC, Blum HE, Thimme R. 2006. Expres-sion of the interleukin-7 receptor alpha chain (CD127) on virus-specific CD8+ T cells identifies function-ally and phenotypically defined memory T cells during acute resolving hepatitis B virus infection. J Virol. 80(7):3532-40.

Boettler T, Spangenberg HC, Neumann-Haefelin C, Panther E, Urbani S, Ferrari C, Blum HE, von Weizsäcker F, Thimme R. 2005. T cells with a CD4+CD25+ regulatory phenotype suppress in vitro pro-liferation of virus-specific CD8+ T cells during chronic HCV infection. J Virol. 79(12):7860-7.

Project in detail +

It is well established that CD4 T cells are central players in orchestrating antiviral humoral and cellular immun-ity. Despite this important role, little is known about the composition and fate of the virus-specific CD4 T cell compartment in viral hepatitis. In the first funding period, we therefore focussed on the longitudinal analysis and functional modification of hepatitis B virus (HBV)- and hepatitis C virus (HCV)-specific CD4 T cells during chronic infection. Specifically, we identified the costimulatory receptor OX40 in combination with the inhibitory receptor PD-1 as potential targets for immune interventions that aim at improving HBV-specific CD4 T cell function (Jacobi/Wild et al., 2019). In addition, we observed dynamic changes of the HCV-specific CD4 T cell phenotype in patients undergoing antiviral therapy with a shift from a predominant Th1 phenotype in the chronic phase of the infection to a Tfh biased phenotype after viral elimination (Smits et al., 2019). However, in contrast to observations in mouse models where virus-specific Th1 cells can sharply be separated from virus-specific Tfh cells, we observed a strong phenotypic overlap between Th1 and Tfh signatures in HCV-specific CD4 T cells. The implications on Th1 and Tfh cell function and the mech-anisms responsible for this phenotypic overlap remain to be identified. Thus, for the second funding period, we propose to continue our studies on antiviral CD4 T cell responses in viral hepatitis, specifically HBV, HCV and hepatitis E virus (HEV) infection. One focus will be on the identification of the lineage commitment of antiviral CD4 T cells in different phases of viral hepatitis in order to identify whether the lack of a distinct lineage commitment is specific for chronic infection or can also be observed in acute and resolved infection (with Thimme, Hofmann, Bengsch, Z03N). Our preliminary results outlined below also suggest that HCV-specific CD4 T cells in chronic infection have an impaired plastic potential to re-differentiate into different lineages. This lack of plasticity could be a consequence of continuous antigen and interferon (IFN) expo-sure in chronic infection as well as metabolic reprogramming. Indeed, in own previous work, we observed that type I IFNs have the potential to potently impact CD4 T cell function. We found that type I IFNs pro-foundly enhance the activation and function of Tfh cells, suggesting that the strong IFN signature in chronic HCV infection might contribute to the expansion of intrahepatic Tfh cells we have previously observed. Therefore, during the next funding period, we will address the role of type I IFNs in modulating CD4 T helper cell responses, especially follicular T helper (Tfh) cell responses (with Hofmann, Knolle and Klingmül-ler/Timmer). By using newly established HCV-specific CD4 T cell clones and primary HCV-specific CD4 T cells, we plan to identify the pathways that are involved in IFN-mediated enhancement of Tfh responses (with Bartenschlager). Specifically, we aim to identify IFN stimulated genes (ISGs) in T cells after culture in the presence of IFN and in patients with chronic viral hepatitis with the goal to analyse the impact of ISGs on Tfh cell activation, metabolism and function (with Bengsch). In addition, we will analyse CD4 T helper cell responses in HEV infection (with Thimme and Neumann-Haefelin). HEV infection is an emerging disease that serves as an excellent model for the analysis of successful immune responses leading to spontaneous viral clearance in immunocompetent hosts. However, spontaneous clearance of HEV does not necessarily pre-vent from re-infection, suggesting a lack of long-lasting, potent neutralizing antibody (nAb) responses. The failure to establish potent, long-lasting humoral immunity has recently been associated with impaired Tfh cell help, particularly help by Tfh cells with a co-existent Th1 signature. Thus, we aim to analyse the T cell re-sponses elicited by HEV infection (with Z03N and Neumann-Haefelin) to analyse whether the breadth and quality of the Tfh response is associated with the nAb responses against HEV (with Dao Thi/Boulant). More-over, we aim to identify MHC class II restricted epitopes from HEV in order to precisely define the viral pro-teins that are targeted and to generate MHC class II tetramers for the analysis of HEV-specific CD4 T cell responses. These analyses will not only give important insights into the role of CD4 T cell responses in HEV infection, but may also be of clinical relevance for vaccine design. Collectively, we aim to identify the ideal Tfh signature that provides the most potent B cell help and to identify factors that influence Tfh differentia-tion and maintenance.

Milestones of the first funding period:
- Identification and comprehensive profiling of Tfh responses following HBV-vaccination and in acute HBV infection in humans
- Analysis of Tfh responses in a HBV mouse model
- Role of Tfh cells in chronic HBV infection