Hepatitis B virus (HBV)-specific CD8 T cells are functionally exhausted in chronic hepatitis B infection, and this condition can be corrected only partially through the modulation of inhibitory pathways, which suggests that a more complex molecular interplay underlies T cell exhaustion. To gain broader insight into this process and identify additional targets for the restoration of T cell function, we compared the transcriptome profiles of HBV-specific CD8 T cells from patients with acute and chronic disease with those of HBV-specific CD8 T cells from patients able to resolve HBV infection spontaneously and influenza (FLU)-specific CD8 T cells from healthy participants. The results indicate that exhausted HBV-specific CD8 T cells are markedly impaired at multiple levels and show substantial downregulation of various cellular processes centered on extensive mitochondrial alterations. A notable improvement of mitochondrial and antiviral CD8 functions was elicited by mitochondrion-targeted antioxidants, which suggests a central role for reactive oxygen species (ROS) in T cell exhaustion. Thus, mitochondria represent promising targets for novel reconstitution therapies to treat chronic hepatitis B infection.
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Gene Expression Omnibus
European Association For The Study Of The Liver. EASL clinical practice guidelines: Management of chronic hepatitis B virus infection. J. Hepatol. 57, 167–185 (2012).
Bertoletti, A. & Ferrari, C. Innate and adaptive immune responses in chronic hepatitis B virus infections: towards restoration of immune control of viral infection. Gut 61, 1754–1764 (2012).
Rehermann, B. & Nascimbeni, M. Immunology of hepatitis B virus and hepatitis C virus infection. Nat. Rev. Immunol. 5, 215–229 (2005).
Barber, D.L. et al. Restoring function in exhausted CD8 T cells during chronic viral infection. Nature 439, 682–687 (2006).
Odorizzi, P.M. & Wherry, E.J. Inhibitory receptors on lymphocytes: insights from infections. J. Immunol. 188, 2957–2965 (2012).
Alatrakchi, N. & Koziel, M. Regulatory T cells and viral liver disease. J. Viral Hepat. 16, 223–229 (2009).
Chen, J.H. et al. Prostaglandin E2 and programmed cell death 1 signaling coordinately impair CTL function and survival during chronic viral infection. Nat. Med. 21, 327–334 (2015).
Ng, C.T. & Oldstone, M.B. IL-10: achieving balance during persistent viral infection. Curr. Top. Microbiol. Immunol. 380, 129–144 (2014).
Veiga-Parga, T., Sehrawat, S. & Rouse, B.T. Role of regulatory T cells during virus infection. Immunol. Rev. 255, 182–196 (2013).
Bengsch, B., Martin, B. & Thimme, R. Restoration of HBV-specific CD8+ T cell function by PD-1 blockade in inactive carrier patients is linked to T cell differentiation. J. Hepatol. 61, 1212–1219 (2014).
Boni, C. et al. Characterization of hepatitis B virus (HBV)-specific T-cell dysfunction in chronic HBV infection. J. Virol. 81, 4215–4225 (2007).
Fisicaro, P. et al. Combined blockade of programmed death-1 and activation of CD137 increase responses of human liver T cells against HBV, but not HCV. Gastroenterology 143, 1576–1585.e4 (2012).
Nebbia, G. et al. Upregulation of the Tim-3/galectin-9 pathway of T cell exhaustion in chronic hepatitis B virus infection. PLoS One 7, e47648 (2012).
Raziorrouh, B. et al. The immunoregulatory role of CD244 in chronic hepatitis B infection and its inhibitory potential on virus-specific CD8+ T-cell function. Hepatology 52, 1934–1947 (2010).
Schurich, A. et al. Role of the coinhibitory receptor cytotoxic T lymphocyte antigen-4 on apoptosis-Prone CD8 T cells in persistent hepatitis B virus infection. Hepatology 53, 1494–1503 (2011).
Tamayo, P. et al. Interpreting patterns of gene expression with self-organizing maps: methods and application to hematopoietic differentiation. Proc. Natl. Acad. Sci. USA 96, 2907–2912 (1999).
Latner, D.R., Kaech, S.M. & Ahmed, R. Enhanced expression of cell cycle regulatory genes in virus-specific memory CD8+ T cells. J. Virol. 78, 10953–10959 (2004).
Wherry, E.J. & Ahmed, R. Memory CD8 T-cell differentiation during viral infection. J. Virol. 78, 5535–5545 (2004).
Pockley, A.G., Muthana, M. & Calderwood, S.K. The dual immunoregulatory roles of stress proteins. Trends Biochem. Sci. 33, 71–79 (2008).
Rowbotham, N.J., Hager-Theodorides, A.L., Furmanski, A.L. & Crompton, T. A novel role for Hedgehog in T-cell receptor signaling: implications for development and immunity. Cell Cycle 6, 2138–2142 (2007).
Tanaka, T., Shibazaki, A., Ono, R. & Kaisho, T. HSP70 mediates degradation of the p65 subunit of nuclear factor κB to inhibit inflammatory signaling. Sci. Signal. 7, ra119 (2014).
Subramanian, A. et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl. Acad. Sci. USA 102, 15545–15550 (2005).
Goldberg, A.L. Functions of the proteasome: from protein degradation and immune surveillance to cancer therapy. Biochem. Soc. Trans. 35, 12–17 (2007).
Chondrogianni, N. et al. Proteasome activation: An innovative promising approach for delaying aging and retarding age-related diseases. Ageing Res. Rev. 23, 37–55 (2015).
Pandita, T.K. ATM function and telomere stability. Oncogene 21, 611–618 (2002).
Sperka, T., Wang, J. & Rudolph, K.L. DNA damage checkpoints in stem cells, ageing and cancer. Nat. Rev. Mol. Cell Biol. 13, 579–590 (2012).
Sena, L.A. et al. Mitochondria are required for antigen-specific T cell activation through reactive oxygen species signaling. Immunity 38, 225–236 (2013).
Buck, M.D., O'Sullivan, D. & Pearce, E.L. T cell metabolism drives immunity. J. Exp. Med. 212, 1345–1360 (2015).
Groettrup, M., Kirk, C.J. & Basler, M. Proteasomes in immune cells: more than peptide producers? Nat. Rev. Immunol. 10, 73–78 (2010).
Kelso, G.F. et al. Selective targeting of a redox-active ubiquinone to mitochondria within cells: antioxidant and antiapoptotic properties. J. Biol. Chem. 276, 4588–4596 (2001).
Dessolin, J. et al. Selective targeting of synthetic antioxidants to mitochondria: towards a mitochondrial medicine for neurodegenerative diseases? Eur. J. Pharmacol. 447, 155–161 (2002).
Devadas, S., Zaritskaya, L., Rhee, S.G., Oberley, L. & Williams, M.S. Discrete generation of superoxide and hydrogen peroxide by T cell receptor stimulation: selective regulation of mitogen-activated protein kinase activation and fas ligand expression. J. Exp. Med. 195, 59–70 (2002).
Harari, A. et al. Functional signatures of protective antiviral T-cell immunity in human virus infections. Immunol. Rev. 211, 236–254 (2006).
McBride, H.M., Neuspiel, M. & Wasiak, S. Mitochondria: more than just a powerhouse. Curr. Biol. 16, R551–R560 (2006).
Smith-Garvin, J.E., Koretzky, G.A. & Jordan, M.S. T cell activation. Annu. Rev. Immunol. 27, 591–619 (2009).
Kumari, S., Curado, S., Mayya, V. & Dustin, M.L. T cell antigen receptor activation and actin cytoskeleton remodeling. Biochim. Biophys. Acta 1838, 546–556 (2014).
Wherry, E.J. et al. Molecular signature of CD8+ T cell exhaustion during chronic viral infection. Immunity 27, 670–684 (2007).
Doering, T.A. et al. Network analysis reveals centrally connected genes and pathways involved in CD8+ T cell exhaustion versus memory. Immunity 37, 1130–1144 (2012).
Kaech, S.M., Hemby, S., Kersh, E. & Ahmed, R. Molecular and functional profiling of memory CD8 T cell differentiation. Cell 111, 837–851 (2002).
Bengsch, B. et al. Bioenergetics insufficiencies due to metabolic alterations regulated by the inhibitory receptor PD-1 are an early driver of CD8+ T cell exhaustion. Immunity 45, 358–373 (2016).
Petrovas, C. et al. Increased mitochondrial mass characterizes the survival defect of HIV-specific CD8+ T cells. Blood 109, 2505–2513 (2007).
López-Otín, C., Blasco, M.A., Partridge, L., Serrano, M. & Kroemer, G. The hallmarks of aging. Cell 153, 1194–1217 (2013).
Ponnappan, S. & Ponnappan, U. Aging and immune function: molecular mechanisms to interventions. Antioxid. Redox Signal. 14, 1551–1585 (2011).
Ron-Harel, N., Sharpe, A.H. & Haigis, M.C. Mitochondrial metabolism in T cell activation and senescence: a mini-review. Gerontology 61, 131–138 (2015).
Sansoni, P. et al. The immune system in extreme longevity. Exp. Gerontol. 43, 61–65 (2008).
O'Sullivan, R.J., Kubicek, S., Schreiber, S.L. & Karlseder, J. Reduced histone biosynthesis and chromatin changes arising from a damage signal at telomeres. Nat. Struct. Mol. Biol. 17, 1218–1225 (2010).
Su, C. et al. DNA damage induces downregulation of histone gene expression through the G1 checkpoint pathway. EMBO J. 23, 1133–1143 (2004).
Cuervo, A.M. & Macian, F. Autophagy and the immune function in aging. Curr. Opin. Immunol. 29, 97–104 (2014).
Szklarczyk, R., Nooteboom, M. & Osiewacz, H.D. Control of mitochondrial integrity in ageing and disease. Phil. Trans. R. Soc. Lond. B 369, 20130439 (2014).
Weber, T.A. & Reichert, A.S. Impaired quality control of mitochondria: aging from a new perspective. Exp. Gerontol. 45, 503–511 (2010).
Carreau, A., El Hafny-Rahbi, B., Matejuk, A., Grillon, C. & Kieda, C. Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia. J. Cell. Mol. Med. 15, 1239–1253 (2011).
Schurich, A. et al. Distinct metabolic requirements of exhausted and functional virus-specific CD8 T cells in the same host. Cell Rep. 16, 1243–1252 (2016).
Dimeloe, S. et al. The immune-metabolic basis of effector memory CD4+ T cell function under hypoxic conditions. J. Immunol. 196, 106–114 (2016).
D'Souza, A.D., Parikh, N., Kaech, S.M. & Shadel, G.S. Convergence of multiple signaling pathways is required to coordinately up-regulate mtDNA and mitochondrial biogenesis during T cell activation. Mitochondrion 7, 374–385 (2007).
van der Windt, G.J.W. et al. Mitochondrial respiratory capacity is a critical regulator of CD8+ T cell memory development. Immunity 36, 68–78 (2012).
Yi, J.S., Holbrook, B.C., Michalek, R.D., Laniewski, N.G. & Grayson, J.M. Electron transport complex I is required for CD8+ T cell function. J. Immunol. 177, 852–862 (2006).
Buck, M.D. et al. Mitochondrial dynamics controls T cell fate through metabolic programming. Cell 166, 63–76 (2016).
Gane, E.J. et al. The mitochondria-targeted anti-oxidant mitoquinone decreases liver damage in a phase II study of hepatitis C patients. Liver Int. 30, 1019–1026 (2010).
Zhang, J. & Fang, H. in Applications of Self-Organizing Maps (ed. Johnsson, M.) 181–204 (In Tech, Rijeka, Croatia, 2012).
Huang, W., Sherman, B.T. & Lempicki, R.A. Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res. 37, 1–13 (2009).
Wang, J., Duncan, D., Shi, Z. & Zhang, B. WEB-based GEne SeT AnaLysis Toolkit (WebGestalt): update 2013. Nucleic Acids Res. 41, W77–W83 (2013).
Warde-Farley, D. et al. The GeneMANIA prediction server: biological network integration for gene prioritization and predicting gene function. Nucleic Acids Res. 38, W214–W20 (2010).
Saito, R. et al. A travel guide to Cytoscape plugins. Nat. Methods 9, 1069–1076 (2012).
We thank M. Murphy (MRC Mitochondrial Biology Unit Wellcome Trust, Cambridge, UK) for the gift of a MitoQ sample; S. Bicciato (Department of Life Sciences, University of Modena and Reggio Emilia, Italy) for introducing us to GSEA; and the Microarray Facility at the University of Ferrara (http://ltta.tecnopoloferrara.it/bioinformatica.php) for help in the initial phase of bioinformatic analysis. We are also grateful to A. Cossarizza (Department of Surgery, Medicine, Dentistry and Morphological Sciences, University of Modena and Reggio Emilia, Italy) and A. Merli (Department of Life Sciences, University of Parma, Italy) for their helpful discussions. This work was supported by a grant from Regione Emilia-Romagna, Italy (Programma di Ricerca Regione-Università 2010–2012; PRUa1RI-2012-006 to C.F.), by a grant (2012.0033 to C.F.) from Fondazione Cassa di Risparmio di Parma (Italy), and by a FIRB grant (RBAP10TPXK to C.F.) from the Italian Ministry of Education, University and Research (MIUR to C.F.).
P.L.: Consultant for BMS, Roche, Gilead Sciences, GSK, MSD
M.L.: Consultant for Gilead, Jansen, BMS, Arbutus, Galapagos, Assembly Pharma, Sanofi/Aventis
C.F.: Consultant for Gilead, Abbvie, Arrowhead
Supplementary Figures 1–8 & Supplementary Table 4 (PDF 1694 kb)
SOM clusters derived from ANOVA filtered genes (XLSX 76 kb)
Functional analysis results related to SOM clusters (XLSX 36 kb)
List of misregulated genes (XLSX 75 kb)
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Fisicaro, P., Barili, V., Montanini, B. et al. Targeting mitochondrial dysfunction can restore antiviral activity of exhausted HBV-specific CD8 T cells in chronic hepatitis B. Nat Med 23, 327–336 (2017). https://doi.org/10.1038/nm.4275
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