T-bet optimizes CD4 T-cell responses against influenza through CXCR3-dependent lung trafficking but not functional programming

Abstract

Although clearance of many intracellular pathogens requires T-bet-dependent CD4 T cell programming, the extent to which T-bet is needed to direct protective CD4 responses against influenza is not known. Here, we characterize wild-type and T-bet-deficient CD4 cells during murine influenza infection. Surprisingly, although T-bet expression has broad impacts on cytokine production by virus-specific CD4 cells, the protective efficacy of T-bet-deficient effector cells is only marginally reduced. This reduction is due to lower CXCR3 expression, leading to suboptimal accumulation of activated T-bet-deficient cells in the infected lung. However, T-bet-deficient cells outcompete wild-type cells to form lung-resident and circulating memory populations following viral clearance, and primed T-bet-deficient mice efficiently clear supralethal heterosubtypic influenza challenges even when depleted of CD8 T cells. These results are relevant to the identification of more incisive correlates of protective T cells and for vaccines that aim to induce durable cellular immunity against influenza.

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References

  1. 1.

    Swain, S. L., McKinstry, K. K. & Strutt, T. M. Expanding roles for CD4( + ) T cells in immunity to viruses. Nat. Rev. Immunol. 12, 136–148 (2012).

  2. 2.

    Szabo, S. J. et al. A novel transcription factor, T-bet, directs Th1 lineage commitment. Cell 100, 655–669 (2000).

  3. 3.

    McKinstry, K. K. et al. Memory CD4 + T cells protect against influenza through multiple synergizing mechanisms. J. Clin. Invest 122, 2847–2856 (2012).

  4. 4.

    Strutt, T. M. et al. Multipronged CD4( + ) T-cell effector and memory responses cooperate to provide potent immunity against respiratory virus. Immunol. Rev. 255, 149–164 (2013).

  5. 5.

    Graham, M. B., Braciale, V. L. & Braciale, T. J. Influenza virus-specific CD4 + T helper type 2 T lymphocytes do not promote recovery from experimental virus infection. J. Exp. Med. 180, 1273–1282 (1994).

  6. 6.

    Brown, D. M., Dilzer, A. M., Meents, D. L. & Swain, S. L. CD4 T cell-mediated protection from lethal influenza: perforin and antibody-mediated mechanisms give a one-two punch. J. Immunol. 177, 2888–2898 (2006).

  7. 7.

    Teijaro, J. R., Verhoeven, D., Page, C. A., Turner, D. & Farber, D. L. Memory CD4 T cells direct protective responses to influenza virus in the lungs through helper-independent mechanisms. J. Virol. 84, 9217–9226 (2010).

  8. 8.

    Brown, D. M., Lee, S., Garcia-Hernandez Mde, L. & Swain, S. L. Multifunctional CD4 cells expressing gamma interferon and perforin mediate protection against lethal influenza virus infection. J. Virol. 86, 6792–6803 (2012).

  9. 9.

    Bot, A., Bot, S. & Bona, C. A. Protective role of gamma interferon during the recall response to influenza virus. J. Virol. 72, 6637–6645 (1998).

  10. 10.

    Graham, M. B. et al. Response to influenza infection in mice with a targeted disruption in the interferon gamma gene. J. Exp. Med. 178, 1725–1732 (1993).

  11. 11.

    Califano, D. et al. IFN-gamma increases susceptibility to influenza A infection through suppression of group II innate lymphoid cells. Mucosal Immunol. 11, 209–219 (2018).

  12. 12.

    Nicol, M. Q. et al. Lack of IFNgamma signaling attenuates spread of influenza A virus in vivo and leads to reduced pathogenesis. Virology 526, 155–164 (2018).

  13. 13.

    McKinstry, K. K. et al. IL-10 deficiency unleashes an influenza-specific Th17 response and enhances survival against high-dose challenge. J. Immunol. 182, 7353–7363 (2009).

  14. 14.

    Eliasson, D. G. et al. M2e-tetramer-specific memory CD4 T cells are broadly protective against influenza infection. Mucosal Immunol. 11, 273–289 (2018).

  15. 15.

    Lord, G. M. et al. T-bet is required for optimal proinflammatory CD4 + T-cell trafficking. Blood 106, 3432–3439 (2005).

  16. 16.

    Strutt, T. M., McKinstry, K. K., Kuang, Y., Bradley, L. M. & Swain, S. L. Memory CD4 + T-cell-mediated protection depends on secondary effectors that are distinct from and superior to primary effectors. Proc. Natl. Acad. Sci. USA 109, E2551–E2560 (2012).

  17. 17.

    Ghosh, S., Chackerian, A. A., Parker, C. M., Ballantyne, C. M. & Behar, S. M. The LFA-1 adhesion molecule is required for protective immunity during pulmonary Mycobacterium tuberculosis infection. J. Immunol. 176, 4914–4922 (2006).

  18. 18.

    Kohlmeier, J. E. et al. CXCR3 directs antigen-specific effector CD4 + T cell migration to the lung during parainfluenza virus infection. J. Immunol. 183, 4378–4384 (2009).

  19. 19.

    Mikhak, Z., Strassner, J. P. & Luster, A. D. Lung dendritic cells imprint T cell lung homing and promote lung immunity through the chemokine receptor CCR4. J. Exp. Med. 210, 1855–1869 (2013).

  20. 20.

    Lazarevic, V., Glimcher, L. H. & Lord, G. M. T-bet: a bridge between innate and adaptive immunity. Nat. Rev. Immunol. 13, 777–789 (2013).

  21. 21.

    Groom, J. R. et al. CXCR3 chemokine receptor-ligand interactions in the lymph node optimize CD4 + T helper 1 cell differentiation. Immunity 37, 1091–1103 (2012).

  22. 22.

    Wang, X. et al. IL-17A Promotes Pulmonary B-1a Cell Differentiation via Induction of Blimp-1 Expression during Influenza Virus Infection. PLoS Pathog. 12, e1005367 (2016).

  23. 23.

    Crowe, C. R. et al. Critical role of IL-17RA in immunopathology of influenza infection. J. Immunol. 183, 5301–5310 (2009).

  24. 24.

    Marshall, H. D. et al. Differential expression of Ly6C and T-bet distinguish effector and memory Th1 CD4(+) cell properties during viral infection. Immunity 35, 633–646 (2011).

  25. 25.

    Hu, Z., Blackman, M. A., Kaye, K. M. & Usherwood, E. J. Functional heterogeneity in the CD4 + T cell response to murine gamma-herpesvirus 68. J. Immunol. 194, 2746–2756 (2015).

  26. 26.

    McKinstry, K. K. et al. Effector CD4 T-cell transition to memory requires late cognate interactions that induce autocrine IL-2. Nat. Commun. 5, 5377 (2014).

  27. 27.

    Dhume, K. & McKinstry, K. K. Early programming and late-acting checkpoints governing the development of CD4 T-cell memory. Immunology 155, 53–62 (2018).

  28. 28.

    Strutt, T. M. et al. IL-15 supports the generation of protective lung-resident memory CD4 T cells. Mucosal Immunol. 11, 668–680 (2018).

  29. 29.

    Zens, K. D. et al. Reduced generation of lung tissue-resident memory T cells during infancy. J. Exp. Med 214, 2915–2932 (2017).

  30. 30.

    Kamperschroer, C., Dibble, J. P., Meents, D. L., Schwartzberg, P. L. & Swain, S. L. SAP is required for Th cell function and for immunity to influenza. J. Immunol. 177, 5317–5327 (2006).

  31. 31.

    Strutt, T. M. et al. Direct IL-6 Signals Maximize Protective Secondary CD4 T Cell Responses against Influenza. J. Immunol. 197, 3260–3270 (2016).

  32. 32.

    Strutt, T. M. et al. Memory CD4 + T cells induce innate responses independently of pathogen. Nat. Med. 16, 558–564 (2010). 551p following 564.

  33. 33.

    Rosas, L. E. et al. Cutting edge: STAT1 and T-bet play distinct roles in determining outcome of visceral leishmaniasis caused by Leishmania donovani. J. Immunol. 177, 22–25 (2006).

  34. 34.

    Harms Pritchard, G. et al. Diverse roles for T-bet in the effector responses required for resistance to infection. J. Immunol. 194, 1131–1140 (2015).

  35. 35.

    Sullivan, B. M. et al. Increased susceptibility of mice lacking T-bet to infection with Mycobacterium tuberculosis correlates with increased IL-10 and decreased IFN-gamma production. J. Immunol. 175, 4593–4602 (2005).

  36. 36.

    Matsuyama, M. et al. Role of Th1/Th17 balance regulated by T-bet in a mouse model of Mycobacterium avium complex disease. J. Immunol. 192, 1707–1717 (2014).

  37. 37.

    Hultgren, O. H., Verdrengh, M. & Tarkowski, A. T-box transcription-factor-deficient mice display increased joint pathology and failure of infection control during staphylococcal arthritis. Microbes Infect. 6, 529–535 (2004).

  38. 38.

    Ravindran, R., Foley, J., Stoklasek, T., Glimcher, L. H. & McSorley, S. J. Expression of T-bet by CD4 T cells is essential for resistance to Salmonella infection. J. Immunol. 175, 4603–4610 (2005).

  39. 39.

    Melillo, A. A., Foreman, O., Bosio, C. M. & Elkins, K. L. T-bet regulates immunity to Francisella tularensis live vaccine strain infection, particularly in lungs. Infect. Immun. 82, 1477–1490 (2014).

  40. 40.

    Cobb, D. et al. T-bet-dependent regulation of CD8 + T-cell expansion during experimental Trypanosoma cruzi infection. Immunology 128, 589–599 (2009).

  41. 41.

    Svensson, A., Nordstrom, I., Sun, J. B. & Eriksson, K. Protective immunity to genital herpes simplex [correction of simpex] virus type 2 infection is mediated by T-bet. J. Immunol. 174, 6266–6273 (2005).

  42. 42.

    Matsui, M., Moriya, O., Yoshimoto, T. & Akatsuka, T. T-bet is required for protection against vaccinia virus infection. J. Virol. 79, 12798–12806 (2005).

  43. 43.

    Lebrun, A. et al. T-bet Is Required for the Rapid Clearance of Attenuated Rabies Virus from Central Nervous System Tissue. J. Immunol. 195, 4358–4368 (2015).

  44. 44.

    Glanville, N. et al. Tbet deficiency causes T helper cell dependent airways eosinophilia and mucus hypersecretion in response to Rhinovirus infection. PLoS Pathog. 12, e1005913 (2016).

  45. 45.

    Yang, Y., Xu, J., Niu, Y., Bromberg, J. S. & Ding, Y. T-bet and eomesodermin play critical roles in directing T cell differentiation to Th1 versus Th17. J. Immunol. 181, 8700–8710 (2008).

  46. 46.

    Lazarevic, V. et al. T-bet represses T(H)17 differentiation by preventing Runx1-mediated activation of the gene encoding RORgammat. Nat. Immunol. 12, 96–104 (2011).

  47. 47.

    Intlekofer, A. M. et al. Anomalous type 17 response to viral infection by CD8 + T cells lacking T-bet and eomesodermin. Science 321, 408–411 (2008).

  48. 48.

    Way, S. S. & Wilson, C. B. Cutting edge: immunity and IFN-gamma production during Listeria monocytogenes infection in the absence of T-bet. J. Immunol. 173, 5918–5922 (2004).

  49. 49.

    Oestreich, K. J. et al. Bcl-6 directly represses the gene program of the glycolysis pathway. Nat. Immunol. 15, 957–964 (2014).

  50. 50.

    van der Windt, G. J. & Pearce, E. L. Metabolic switching and fuel choice during T-cell differentiation and memory development. Immunol. Rev. 249, 27–42 (2012).

  51. 51.

    Er J. Z., Koean R. A. G., Ding J. L. Loss of T-bet confers survival advantage to influenza-bacterial superinfection. EMBO J. 38, (2018).

  52. 52.

    Fang, D. et al. Transient T-bet expression functionally specifies a distinct T follicular helper subset. J. Exp. Med. 215, 2705–2714 (2018).

  53. 53.

    Knox, J. J., Myles, A. & Cancro, M. P. T-bet(+) memory B cells: generation, function, and fate. Immunol. Rev. 288, 149–160 (2019).

  54. 54.

    McKinstry, K. K. et al. Rapid default transition of CD4 T cell effectors to functional memory cells. J. Exp. Med. 204, 2199–2211 (2007).

  55. 55.

    Sell, S. et al. Intraepithelial T-cell cytotoxicity, induced bronchus-associated lymphoid tissue, and proliferation of pneumocytes in experimental mouse models of influenza. Viral Immunol. 27, 484–496 (2014).

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Acknowledgements

We thank the NIH Tetramer Core Facility for providing the NP311–235 tetramer and control reagents. We thank the University of Central Florida’s Vivarium staff for providing excellent care for the animals in this study. We thank Dr. Priyadharshini Devarajan for helpful discussions. This work was supported by American Heart Association grant 14SDG18600020 (to K.K.M.), National Institutes of Health Grant AI117457 (to T.M.S.) and by funds provided by the University of Central Florida.

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K.D. and C.F. performed all experiments. K.D. and K.K.M. analyzed data and wrote the manuscript. S.S. performed blinded analysis of histopathology. T.M.S. provided key reagents, reviewed and critiqued the manuscript and figures, and contributed to interpretation of the findings.

Correspondence to K. Kai McKinstry.

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Dhume, K., Finn, C.M., Strutt, T.M. et al. T-bet optimizes CD4 T-cell responses against influenza through CXCR3-dependent lung trafficking but not functional programming. Mucosal Immunol 12, 1220–1230 (2019). https://doi.org/10.1038/s41385-019-0183-z

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