Memory CD4+ T cells induce innate responses independently of pathogen

Journal name:
Nature Medicine
Volume:
16,
Pages:
558–564
Year published:
DOI:
doi:10.1038/nm.2142
Received
Accepted
Published online

Abstract

Inflammation induced by recognition of pathogen-associated molecular patterns markedly affects subsequent adaptive responses. We asked whether the adaptive immune system can also affect the character and magnitude of innate inflammatory responses. We found that the response of memory, but not naive, CD4+ T cells enhances production of multiple innate inflammatory cytokines and chemokines (IICs) in the lung and that, during influenza infection, this leads to early control of virus. Memory CD4+ T cell–induced IICs and viral control require cognate antigen recognition and are optimal when memory cells are either T helper type 1 (TH1) or TH17 polarized but are independent of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) production and do not require activation of conserved pathogen recognition pathways. This represents a previously undescribed mechanism by which memory CD4+ T cells induce an early innate response that enhances immune protection against pathogens.

At a glance

Figures

  1. Memory CD4+ T cells induce an acute increase in IICs upon influenza infection.
    Figure 1: Memory CD4+ T cells induce an acute increase in IICs upon influenza infection.

    (a) IIC concentrations 40 h after challenge in naive C57BL/6 mice, or mice primed with influenza A/Phil 60 d before treatment, treated with isotype, CD4- or Thy1.2-depleting antibody before influenza A/PR8 challenge (n = 5 mice per group). (b) IIC concentrations after bulk CD4+ T cells were isolated from naive or influenza A/PR8–primed mice (polyclonal memory) and equal numbers transferred to naive hosts or, alternatively, naive or in vivo– or in vitro–generated HNT memory cells were adoptively transferred to naive BALB/c hosts. All recipients were challenged with influenza A/PR8 and lung homogenates assessed for IIC after 40 h (n = 5 mice per group). Dotted lines in all figures represent levels of IICs in the absence of infection. Error bars indicate s.d.; *P < 0.05, **P < 0.005 (one-way analysis of variance (ANOVA) followed by Bonferroni's post hoc test).

  2. Role of IFN-[gamma], TNF-[alpha] and CCL3 in IIC upregulation by memory CD4+ T cells.
    Figure 2: Role of IFN-γ, TNF-α and CCL3 in IIC upregulation by memory CD4+ T cells.

    (a) Fold increase in IFN-γ detected in lung homogenate with transfer of TH1-polarized memory versus naive Ifng−/− CD4+ T cells to unprimed mice, on days 2 and 3 after infection with influenza A/PR8 (n = 5 mice per group). (b) Staining of lung cells for the indicated surface markers to identify IFN-γ–producing cells (EYFP+) after memory OT-II cells transferred to unprimed C57BL/6 Yeti hosts (n = 5 mice per group). NK, natural killer; flu, influenza. (c,d) Naive or TH1-polarized memory OT-II cells were transferred to WT, Ifngr−/−, Tnfrsf1ab−/− or Ccr5−/− hosts and then infected with influenza A/PR8. (c) Concentrations of IICs at 40 h after infection. (d) Viral titer on day 4 after infection (n = 5 mice per d). Error bars indicate s.d.; *P < 0.05, **P < 0.005, ***P < 0.001 (Student's t test). ND, not detected.

  3. TH1- or TH17-polarization is required for enhanced IIC response and viral control.
    Figure 3: TH1- or TH17-polarization is required for enhanced IIC response and viral control.

    Naive, TH1-, TH17- or TH2-polarized or TH0 unpolarized memory HNT cells were transferred to BALB/c hosts, which were then infected with influenza A/PR8. (a) Concentrations of IICs in lungs 40 h after infection (n = 5 mice per group). (b) Pulmonary viral titers (n = 5 mice per group). Error bars indicate s.d.; *P < 0.05, **P < 0.005, ***P < 0.001 (one-way ANOVA followed by Bonferroni's post hoc test).

  4. Recognition of antigen in the lung is sufficient for IIC upregulation.
    Figure 4: Recognition of antigen in the lung is sufficient for IIC upregulation.

    CFSE-labeled Thy-disparate naive or TH1-polarized memory HNT cells were transferred to separate hosts, which were then infected with influenza A/PR8 (n = 5 per group). (a) Numbers of donor cells in spleen, draining lymph node (DLN) and lung 40 h after infection. (b) Representative CFSE and CD69 expression of donor cells 2 and 6 d after infection. Naive or memory OT-II cells were transferred to sham-treated or splenectomized Lta−/− hosts and infected with influenza A/PR8-OVAII. (c) Pulmonary IIC concentrations at 40 h after infection (n = 5 mice per group). (d) Pulmonary viral titers (n = 5 per day). Error bars indicate s.d.; *P < 0.05, **P < 0.005, ***P < 0.001 (one-way ANOVA followed by Bonferroni's post hoc test or t test).

  5. Cognate recognition of antigen on MHC class II-expressing CD11c+ cells is sufficient to induce IIC upregulation.
    Figure 5: Cognate recognition of antigen on MHC class II–expressing CD11c+ cells is sufficient to induce IIC upregulation.

    (a) The ratio of viral titers on day 4 in naive C57/BL6 mice receiving naive or TH1-polarized OT-II memory cells and then infected with influenza A/PR8 or A/PR8-OVAII. (n = 5 mice per group per virus.) (b,c) Concentrations of IICs in lung homogenates 40 h after influenza A/PR8-OVAII infection in WT, H2-Ab1−/− or CD11c Tg.H2-Ab1−/− mice receiving TH1-polarized memory OT-II cells (n = 5 mice per group). (b) and pulmonary viral titers (n = 5 mice per group) (c). (d,e) Number of CD11c+ cells in the lung 40 h after infection in mice receiving naive or memory cells (d) and (e) expression of MHC-II, CD40 and CD80 on CD11c+ cells (e). (f) Expression of MHC-II and CD40 on DCs cultured with memory cells. Anti-CD3, CD3-specific antibody. (g) Cytokines detected in 48-h supernatants of naive or memory HNT cells cultured with DCs. Error bars indicate s.d.; *P < 0.05, **P < 0.005, ***P < 0.001 (one-way ANOVA followed by Bonferroni's post hoc test or t test).

  6. Memory CD4+ T cells induce IIC responses independently of PAMP recognition.
    Figure 6: Memory CD4+ T cells induce IIC responses independently of PAMP recognition.

    (a,b) Naive or TH1-polarized OT-II memory cells were transferred to naive WT, Ifnar2−/− or Myd88−/−;Ticam1 hosts, which were then infected with influenza A/PR8-OVAII. (a) Concentrations of IICs detected in lungs 40 h after infection (n = 5 mice per group). (b) Viral titers. (c) Concentrations of IICs 40 h after naive or TH1-polarized memory OT-II cells were transferred to C57BL/6 hosts, which were then administered 100 or 10 μg of soluble LPS-free OVA intranasally (i.n.) (n = 5 mice per group). (d) Memory OT-II cells were transferred to C57/BL6 hosts administered LPS-free OVA as in c and infected with A/PR8 on the same day or 7 d later, or only infected with A/PR8, and viral titers determined (n = 5 per group). Error bars indicate s.d.; *P < 0.05, **P < 0.005, ***P < 0.001 (one-way ANOVA followed by Bonferroni's post hoc test or t test).

References

  1. Janeway, C.A. Jr. & Medzhitov, R. Innate immune recognition. Annu. Rev. Immunol. 20, 197216 (2002).
  2. Iwasaki, A. & Medzhitov, R. Toll-like receptor control of the adaptive immune responses. Nat. Immunol. 5, 987995 (2004).
  3. Pulendran, B. Modulating vaccine responses with dendritic cells and Toll-like receptors. Immunol. Rev. 199, 227250 (2004).
  4. Pulendran, B., Palucka, K. & Banchereau, J. Sensing pathogens and tuning immune responses. Science 293, 253256 (2001).
  5. Powell, T.J. et al. Priming with cold-adapted influenza A does not prevent infection but elicits long-lived protection against supralethal challenge with heterosubtypic virus. J. Immunol. 178, 10301038 (2007).
  6. Swain, S.L. et al. CD4+ T-cell memory: generation and multi-faceted roles for CD4+ T cells in protective immunity to influenza. Immunol. Rev. 211, 822 (2006).
  7. Rogers, P.R., Dubey, C. & Swain, S.L. Qualitative changes accompany memory T cell generation: faster, more effective responses at lower doses of antigen. J. Immunol. 164, 23382346 (2000).
  8. London, C.A., Lodge, M.P. & Abbas, A.K. Functional responses and costimulator dependence of memory CD4+ T cells. J. Immunol. 164, 265272 (2000).
  9. Bradley, L.M., Duncan, D.D., Yoshimoto, K. & Swain, S.L. Memory effectors: a potent, IL-4–secreting helper T cell population that develops in vivo after restimulation with antigen. J. Immunol. 150, 31193130 (1993).
  10. Dahl, M.E., Dabbagh, K., Liggitt, D., Kim, S. & Lewis, D.B. Viral-induced T helper type 1 responses enhance allergic disease by effects on lung dendritic cells. Nat. Immunol. 5, 337343 (2004).
  11. Didierlaurent, A. et al. Sustained desensitization to bacterial Toll-like receptor ligands after resolution of respiratory influenza infection. J. Exp. Med. 205, 323329 (2008).
  12. Chace, J.H., Cowdery, J.S. & Field, E.H. Effect of anti-CD4 on CD4 subsets. I. Anti-CD4 preferentially deletes resting, naive CD4 cells and spares activated CD4 cells. J. Immunol. 152, 405412 (1994).
  13. Scott, B. et al. A role for non-MHC genetic polymorphism in susceptibility to spontaneous autoimmunity. Immunity 1, 7383 (1994).
  14. McKinstry, K.K. et al. Rapid default transition of CD4 T cell effectors to functional memory cells. J. Exp. Med. 204, 21992211 (2007).
  15. Thomas, P.G. et al. An unexpected antibody response to an engineered influenza virus modifies CD8+ T cell responses. Proc. Natl. Acad. Sci. USA 103, 27642769 (2006).
  16. Mayer, K.D. et al. The functional heterogeneity of type 1 effector T cells in response to infection is related to the potential for IFN-γ production. J. Immunol. 174, 77327739 (2005).
  17. Boehm, U., Klamp, T., Groot, M. & Howard, J.C. Cellular responses to interferon-gamma. Annu. Rev. Immunol. 15, 749795 (1997).
  18. Farber, J.M. Mig and IP-10: CXC chemokines that target lymphocytes. J. Leukoc. Biol. 61, 246257 (1997).
  19. Amichay, D. et al. Genes for chemokines MuMig and Crg-2 are induced in protozoan and viral infections in response to IFN-γ with patterns of tissue expression that suggest nonredundant roles in vivo . J. Immunol. 157, 45114520 (1996).
  20. Nakanishi, Y., Lu, B., Gerard, C. & Iwasaki, A. CD8+ T lymphocyte mobilization to virus-infected tissue requires CD4+ T-cell help. Nature 462, 510513 (2009).
  21. 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, 28882898 (2006).
  22. McKinstry, K.K. et al. IL-10 deficiency unleashes an influenza-specific TH17 response and enhances survival against high-dose challenge. J. Immunol. 182, 73537363 (2009).
  23. Wakim, L.M., Waithman, J., van Rooijen, N., Heath, W.R. & Carbone, F.R. Dendritic cell-induced memory T cell activation in nonlymphoid tissues. Science 319, 198202 (2008).
  24. De Togni, P. et al. Abnormal development of peripheral lymphoid organs in mice deficient in lymphotoxin. Science 264, 703707 (1994).
  25. Debbabi, H. et al. Primary type II alveolar epithelial cells present microbial antigens to antigen-specific CD4+ T cells. Am. J. Physiol. Lung Cell Mol. Physiol. 289, L274L279 (2005).
  26. Lemos, M.P., Fan, L., Lo, D. & Laufer, T.M. CD8α+ and CD11b+ dendritic cell–restricted MHC class II controls TH1 CD4+ T cell immunity. J. Immunol. 171, 50775084 (2003).
  27. Diebold, S.S., Kaisho, T., Hemmi, H., Akira, S. & Reis e Sousa, C. Innate antiviral responses by means of TLR7-mediated recognition of single-stranded RNA. Science 303, 15291531 (2004).
  28. Imai, Y. et al. Identification of oxidative stress and Toll-like receptor 4 signaling as a key pathway of acute lung injury. Cell 133, 235249 (2008).
  29. Pichlmair, A. et al. RIG-I–mediated antiviral responses to single-stranded RNA bearing 5′-phosphates. Science 314, 9971001 (2006).
  30. Guarda, G. et al. T cells dampen innate immune responses through inhibition of NLRP1 and NLRP3 inflammasomes. Nature 460, 269273 (2009).
  31. Kim, K.D. et al. Adaptive immune cells temper initial innate responses. Nat. Med. 13, 12481252 (2007).
  32. Salomon, R., Hoffmann, E. & Webster, R.G. Inhibition of the cytokine response does not protect against lethal H5N1 influenza infection. Proc. Natl. Acad. Sci. USA 104, 1247912481 (2007).
  33. Tuvim, M.J., Evans, S.E., Clement, C.G., Dickey, B.F. & Gilbert, B.E. Augmented lung inflammation protects against influenza A pneumonia. PLoS One 4, e4176 (2009).
  34. Kalinski, P. & Moser, M. Consensual immunity: success-driven development of T-helper-1 and T-helper-2 responses. Nat. Rev. Immunol. 5, 251260 (2005).
  35. Hale, B.G., Randall, R.E., Ortin, J. & Jackson, D. The multifunctional NS1 protein of influenza A viruses. J. Gen. Virol. 89, 23592376 (2008).
  36. Joffre, O., Nolte, M.A., Sporri, R. & Reis e Sousa, C. Inflammatory signals in dendritic cell activation and the induction of adaptive immunity. Immunol. Rev. 227, 234247 (2009).
  37. Dienz, O. et al. The induction of antibody production by IL-6 is indirectly mediated by IL-21 produced by CD4+ T cells. J. Exp. Med. 206, 6978 (2009).
  38. Szretter, K.J. et al. Role of host cytokine responses in the pathogenesis of avian H5N1 influenza viruses in mice. J. Virol. 81, 27362744 (2007).
  39. Schmitz, N., Kurrer, M., Bachmann, M.F. & Kopf, M. Interleukin-1 is responsible for acute lung immunopathology but increases survival of respiratory influenza virus infection. J. Virol. 79, 64416448 (2005).
  40. Lee, S.W., Youn, J.W., Seong, B.L. & Sung, Y.C. IL-6 induces long-term protective immunity against a lethal challenge of influenza virus. Vaccine 17, 490496 (1999).
  41. Hama, Y. et al. Interleukin 12 is a primary cytokine responding to influenza virus infection in the respiratory tract of mice. Acta Virol. 53, 233240 (2009).
  42. GeurtsvanKessel, C.H. & Lambrecht, B.N. Division of labor between dendritic cell subsets of the lung. Mucosal Immunol. 1, 442450 (2008).
  43. McGill, J., Heusel, J.W. & Legge, K.L. Innate immune control and regulation of influenza virus infections. J. Leukoc. Biol. 86, 803812 (2009).
  44. Monteiro, J.M., Harvey, C. & Trinchieri, G. Role of interleukin-12 in primary influenza virus infection. J. Virol. 72, 48254831 (1998).
  45. Zhao, M.Q. et al. Alveolar epithelial cell chemokine expression triggered by antigen-specific cytolytic CD8+ T cell recognition. J. Clin. Invest. 106, R49R58 (2000).
  46. Guidotti, L.G. & Chisari, F.V. Noncytolytic control of viral infections by the innate and adaptive immune response. Annu. Rev. Immunol. 19, 6591 (2001).
  47. Le Saout, C., Mennechet, S., Taylor, N. & Hernandez, J. Memory-like CD8+ and CD4+ T cells cooperate to break peripheral tolerance under lymphopenic conditions. Proc. Natl. Acad. Sci. USA 105, 1941419419 (2008).
  48. Elyaman, W. et al. Distinct functions of autoreactive memory and effector CD4+ T cells in experimental autoimmune encephalomyelitis. Am. J. Pathol. 173, 411422 (2008).
  49. Latham, K.A., Whittington, K.B., Zhou, R., Qian, Z. & Rosloniec, E.F. Ex vivo characterization of the autoimmune T cell response in the HLA-DR1 mouse model of collagen-induced arthritis reveals long-term activation of type II collagen-specific cells and their presence in arthritic joints. J. Immunol. 174, 39783985 (2005).
  50. Strutt, T.M., Uzonna, J., McKinstry, K.K. & Bretscher, P.A. Activation of thymic T cells by MHC alloantigen requires syngeneic, activated CD4+ T cells and B cells as APC. Int. Immunol. 18, 719728 (2006).

Download references

Author information

  1. These authors contributed equally to this work.

    • Tara M Strutt &
    • K Kai McKinstry

Affiliations

  1. Trudeau Institute, Saranac Lake, New York, USA.

    • Tara M Strutt,
    • K Kai McKinstry,
    • John P Dibble,
    • Caylin Winchell,
    • Yi Kuang,
    • Jonathan D Curtis,
    • Gail Huston,
    • Richard W Dutton &
    • Susan L Swain

Contributions

T.M.S. and K.K.M. contributed equally to the design, processing, collection and analysis of data and, together with S.L.S., wrote the paper. S.L.S. and R.W.D. contributed to study design. J.P.D., C.W., Y.K., J.D.C. and G.H. processed and collected data. All authors discussed results and commented on the manuscript.

Competing financial interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to:

Author details

Supplementary information

PDF files

  1. Supplementary Text and Figures (864K)

    Supplementary Figures 1–5

Additional data