Article | Published:

IRF4-dependent dendritic cells regulate CD8+ T-cell differentiation and memory responses in influenza infection

Mucosal Immunology (2019) | Download Citation



Acute respiratory disease caused by influenza viruses is imperfectly mitigated by annual vaccination to select strains. Development of vaccines that elicit lung-resident memory CD8+ T cells (TRM) would offer more universal protection to seasonal and emerging pandemic viruses. Understanding how lung-resident dendritic cells (DCs) regulate TRM differentiation would be an important step in this process. Here, we used CD11c-cre-Irf4f/f (KO) mice, which lack lung-resident IRF4-dependent CD11b+CD24hi DCs and show IRF4 deficiency in other lung cDC subsets, to determine if IRF4-expressing DCs regulate CD8+ memory precursor cells and TRM during influenza A virus (IAV) infection. KO mice showed defective CD8+ T-cell memory, stemming from a deficit of T regulatory cells and memory precursor cells with decreased Foxo1 expression. Transfer of wild-type CD11b+CD24hi DCs into KO mice restored CD8+ memory precursor cell numbers to wild-type levels. KO mice recovered from a primary infection harbored reduced numbers of CD8+ TRM and showed deficient expansion of IFNγ+CD8+ T cells and increased lung pathology upon challenge with heterosubtypic IAV. Thus, vaccination strategies that harness the function of IRF4-dependent DCs could promote the differentiation of CD8+ TRM during IAV infection.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


  1. 1.

    Braciale, T. J., Sun, J. & Kim, T. S. Regulating the adaptive immune response to respiratory virus infection. Nat. Rev. Immunol. 12, 295–305 (2012).

  2. 2.

    Moskophidis, D. & Kioussis, D. Contribution of virus-specific CD8 + cytotoxic T cells to virus clearance or pathologic manifestations of influenza virus infection in a T cell receptor transgenic mouse model. J. Exp. Med 188, 223–232 (1998).

  3. 3.

    Krammer, F. et al. Influenza. Nat. Rev. Dis. Prim. 4, 3 (2018).

  4. 4.

    Zens, K. D., Chen, J. K. & Farber, D. L. Vaccine-generated lung tissue-resident memory T cells provide heterosubtypic protection to influenza infection. JCI Insight 1, e85832 (2016).

  5. 5.

    Wakim, L. M., Smith, J., Caminschi, I., Lahoud, M. H. & Villadangos, J. A. Antibody-targeted vaccination to lung dendritic cells generates tissue-resident memory CD8 T cells that are highly protective against influenza virus infection. Mucosal Immunol. 8, 1060–1071 (2015).

  6. 6.

    Bajaña, S., Turner, S., Paul, J., Ainsua-Enrich, E. & Kovats, S. IRF4 and IRF8 act in CD11c+ cells to regulate terminal differentiation of lung tissue dendritic cells. J. Immunol. 196, 1666–1677 (2016).

  7. 7.

    Schlitzer, A. et al. IRF4 transcription factor-dependent CD11b+ dendritic cells in human and mouse control mucosal IL-17 cytokine responses. Immunity 38, 970–983 (2013).

  8. 8.

    Edelson, B. T. et al. Peripheral CD103+ dendritic cells form a unified subset developmentally related to CD8alpha+ conventional dendritic cells. J. Exp. Med. 207, 823–836 (2010).

  9. 9.

    Ginhoux, F. et al. The origin and development of nonlymphoid tissue CD103+ DCs. J. Exp. Med. 206, 3115–3130 (2009).

  10. 10.

    Haniffa, M. et al. Human tissues contain CD141hi cross-presenting dendritic cells with functional homology to mouse CD103+ nonlymphoid dendritic cells. Immunity 37, 60–73 (2012).

  11. 11.

    Nakano, H. et al. Blood-derived inflammatory dendritic cells in lymph nodes stimulate acute T helper type 1 immune responses. Nat. Immunol. 10, 394–402 (2009).

  12. 12.

    Aldridge, J. R. J. et al. TNF/iNOS-producing dendritic cells are the necessary evil of lethal influenza virus infection. Proc. Natl Acad. Sci. USA 106, 5306–5311 (2009).

  13. 13.

    Kim, T. S. & Braciale, T. J. Respiratory dendritic cell subsets differ in their capacity to support the induction of virus-specific cytotoxic CD8+ T cell responses. PLoS ONE 4, e4204 (2009).

  14. 14.

    GeurtsvanKessel, C. H. et al. Clearance of influenza virus from the lung depends on migratory langerin+ CD11b- but not plasmacytoid dendritic cells. J. Exp. Med. 205, 1621–1634 (2008).

  15. 15.

    Ballesteros-Tato, A., Leon, B., Lund, F. E. & Randall, T. D. Temporal changes in dendritic cell subsets, cross-priming and costimulation via CD70 control CD8(+) T cell responses to influenza. Nat. Immunol. 11, 216–224 (2010).

  16. 16.

    Ho, A. W. et al. Lung CD103+ dendritic cells efficiently transport influenza virus to the lymph node and load viral antigen onto MHC class I for presentation to CD8 T cells. J. Immunol. 187, 6011–6021 (2011).

  17. 17.

    Helft, J. et al. Cross-presenting CD103+ dendritic cells are protected from influenza virus infection. J. Clin. Invest. 122, 4037–4047 (2012).

  18. 18.

    Krishnaswamy, J. K. et al. Migratory CD11b+ conventional dendritic cells induce T follicular helper cell-dependent antibody responses. Sci. Immunol. 2, pii: eaam9169 (2017).

  19. 19.

    Kim, T. S., Gorski, S. A., Hahn, S., Murphy, K. M. & Braciale, T. J. Distinct dendritic cell subsets dictate the fate decision between effector and memory CD8(+) T cell differentiation by a CD24-dependent mechanism. Immunity 40, 400–413 (2014).

  20. 20.

    Martínez-López, M., Iborra, S., Conde-Garrosa, R. & Sancho, D. Batf3-dependent CD103+ dendritic cells are major producers of IL-12 that drive local Th1 immunity against Leishmania major infection in mice. Eur. J. Immunol. 45, 119–129 (2015).

  21. 21.

    Williams, J. W. et al. Transcription factor IRF4 drives dendritic cells to promote Th2 differentiation. Nat. Commun. 4, 2990 (2013).

  22. 22.

    Rosato, P. C., Beura, L. K. & Masopust, D. Tissue resident memory T cells and viral immunity. Curr. Opin. Virol. 22, 44–50 (2017).

  23. 23.

    Gebhardt, T., Mueller, S. N., Heath, W. R. & Carbone, F. R. Peripheral tissue surveillance and residency by memory T cells. Trends Immunol. 34, 27–32 (2013).

  24. 24.

    Wu, T. et al. Lung-resident memory CD8 T cells (TRM) are indispensable for optimal cross-protection against pulmonary virus infection. J. Leukoc. Biol. 95, 215–224 (2014).

  25. 25.

    McMaster, S. R., Wilson, J. J., Wang, H. & Kohlmeier, J. E. Airway-resident memory CD8 T cells provide antigen-specific protection against respiratory virus challenge through rapid IFN-γ production. J. Immunol. 195, 203–209 (2015).

  26. 26.

    Slütter, B. et al. Dynamics of influenza-induced lung-resident memory T cells underlie waning heterosubtypic immunity. Sci. Immunol. 2, pii: eaag2031 (2017).

  27. 27.

    Shane, H. L., Reagin, K. L. & Klonowski, K. D. The respiratory environment diverts the development of antiviral memory CD8 T cells. J. Immunol. 200, 3752–3761 (2018).

  28. 28.

    Purwar, R. et al. Resident memory T cells (T(RM)) are abundant in human lung: diversity, function, and antigen specificity. PLoS ONE 6, e16245 (2011).

  29. 29.

    Kumar, B. V. et al. Human tissue-resident memory T cells are defined by core transcriptional and functional signatures in lymphoid and mucosal sites. Cell Rep. 20, 2921–2934 (2017).

  30. 30.

    Iborra, S. et al. Optimal generation of tissue-resident but not circulating memory T cells during viral infection requires crosspriming by DNGR-1+ dendritic cells. Immunity 45, 847–860 (2016).

  31. 31.

    Kaech, S. M. & Cui, W. Transcriptional control of effector and memory CD8+ T cell differentiation. Nat. Rev. Immunol. 12, 749–761 (2012).

  32. 32.

    Chang, J. T., Wherry, E. J. & Goldrath, A. W. Molecular regulation of effector and memory T cell differentiation. Nat. Immunol. 15, 1104–1115 (2014).

  33. 33.

    Mackay, L. K. & Kallies, A. Transcriptional regulation of tissue-resident lymphocytes. Trends Immunol. 38, 94–103 (2017).

  34. 34.

    Laidlaw, B. J. et al. Production of IL-10 by CD4(+) regulatory T cells during the resolution of infection promotes the maturation of memory CD8(+) T cells. Nat. Immunol. 16, 871–879 (2015).

  35. 35.

    Croom, H. A. et al. Memory precursor phenotype of CD8+ T cells reflects early antigenic experience rather than memory numbers in a model of localized acute influenza infection. Eur. J. Immunol. 41, 682–693 (2011).

  36. 36.

    Betts, R. J. et al. Influenza A virus infection results in a robust, antigen-responsive, and widely disseminated Foxp3+ regulatory T cell response. J. Virol. 86, 2817–2825 (2012).

  37. 37.

    Akbari, M. et al. IRF4 in dendritic cells inhibits IL-12 production and controls Th1 immune responses against Leishmania major. J. Immunol. 192, 2271–2279 (2014).

  38. 38.

    Rao, R. R., Li, Q., Gubbels Bupp, M. R. & Shrikant, P. A. Transcription factor Foxo1 represses T-bet-mediated effector functions and promotes memory CD8(+) T cell differentiation. Immunity 36, 374–387 (2012).

  39. 39.

    Hess Michelini, R., Doedens, A. L., Goldrath, A. W. & Hedrick, S. M. Differentiation of CD8 memory T cells depends on Foxo1. J. Exp. Med. 210, 1189–1200 (2013).

  40. 40.

    Carlson, C. M. et al. Kruppel-like factor 2 regulates thymocyte and T-cell migration. Nature 442, 299–302 (2006).

  41. 41.

    Li, M. O. & Flavell, R. A. Contextual regulation of inflammation: a duet by transforming growth factor-beta and interleukin-10. Immunity 28, 468–476 (2008).

  42. 42.

    Kreijtz, J. H. et al. Primary influenza A virus infection induces cross-protective immunity against a lethal infection with a heterosubtypic virus strain in mice. Vaccine 25, 612–620 (2007).

  43. 43.

    Tejera, M. M., Kim, E. H., Sullivan, J. A., Plisch, E. H. & Suresh, M. FoxO1 controls effector-to-memory transition and maintenance of functional CD8 T cell memory. J. Immunol. 191, 187–199 (2013).

  44. 44.

    Delpoux, A. et al. Continuous activity of Foxo1 is required to prevent anergy and maintain the memory state of CD8+ T cells. J. Exp. Med. 215, 575–594 (2018).

  45. 45.

    Kim, E. H. & Suresh, M. Role of PI3K/Akt signaling in memory CD8 T cell differentiation. Front. Immunol. 4, 20 (2013).

  46. 46.

    Chen, W. et al. Conversion of peripheral CD4+ CD25- naive T cells to CD4+ CD25+ regulatory T cells by TGF-beta induction of transcription factor Foxp3. J. Exp. Med. 198, 1875–1886 (2003).

  47. 47.

    Jelley-Gibbs, D. M. et al. Unexpected prolonged presentation of influenza antigens promotes CD4 T cell memory generation. J. Exp. Med. 202, 697–706 (2005).

  48. 48.

    Chang, Y. H. et al. Dichotomous expression of TNF superfamily ligands on antigen-presenting cells controls post-priming anti-viral CD4+ T cell immunity. Immunity 47, 943–958.e9 (2017).

  49. 49.

    León, B., Ballesteros-Tato, A., Randall, T. D. & Lund, F. E. Prolonged antigen presentation by immune complex-binding dendritic cells programs the proliferative capacity of memory CD8 T cells. J. Exp. Med. 211, 1637–1655 (2014).

  50. 50.

    Desai, P., Tahiliani, V., Stanfield, J., Abboud, G. & Salek-Ardakani, S. Inflammatory monocytes contribute to the persistence of CXCR3hi CX3CR1lo circulating and lung-resident memory CD8+ T cells following respiratory virus infection. Immunol. Cell Biol. 96, 370–378 (2018).

  51. 51.

    Shin, H., Kumamoto, Y., Gopinath, S. & Iwasaki, A. CD301b+ dendritic cells stimulate tissue-resident memory CD8+ T cells to protect against genital HSV-2. Nat. Commun. 7, 13346 (2016).

  52. 52.

    Park, S. L. et al. Local proliferation maintains a stable pool of tissue-resident memory T cells after antiviral recall responses. Nat. Immunol. 19, 183–191 (2018).

  53. 53.

    Beura, L. K. et al. Intravital mucosal imaging of CD8+ resident memory T cells shows tissue-autonomous recall responses that amplify secondary memory. Nat. Immunol. 19, 173–182 (2018).

  54. 54.

    Klein, U. et al. Transcription factor IRF4 controls plasma cell differentiation and class-switch recombination. Nat. Immunol. 7, 773–782 (2006).

  55. 55.

    Caton, M. L., Smith-Raska, M. R. & Reizis, B. Notch-RBP-J signaling controls the homeostasis of CD8- dendritic cells in the spleen. J. Exp. Med. 204, 1653–1664 (2007).

  56. 56.

    Anderson, K. G. et al. Intravascular staining for discrimination of vascular and tissue leukocytes. Nat. Protoc. 9, 209–222 (2014).

  57. 57.

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

Download references


We thank Dr. Gillian Air and Shelly Gulati at OUHSC for assistance with growing and titering virus in eggs, Dr. Diana Hamilton and Jacob Bass in the OMRF Flow Cytometry Core Facility, and the staff of the Comparative Medicine Facility. This work was supported by NIH HL119501 (to S. Kovats) and by an OMRF Patricia and Don Capra Predoctoral Fellowship (to S. Kadel).

Author information


  1. Arthritis & Clinical Immunology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, 73104, USA

    • Erola Ainsua-Enrich
    • , Ibrahim Hatipoglu
    • , Sapana Kadel
    • , Sean Turner
    • , Jinny Paul
    • , Simar Singh
    • , Harini Bagavant
    •  & Susan Kovats
  2. Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA

    • Sapana Kadel
    •  & Susan Kovats


  1. Search for Erola Ainsua-Enrich in:

  2. Search for Ibrahim Hatipoglu in:

  3. Search for Sapana Kadel in:

  4. Search for Sean Turner in:

  5. Search for Jinny Paul in:

  6. Search for Simar Singh in:

  7. Search for Harini Bagavant in:

  8. Search for Susan Kovats in:


E.A.-E. designed and performed experiments, analyzed data and wrote the manuscript. I.H. and H.B. designed and performed experiments and analyzed data. S. Kadel, S.T., J.P. and S.S. performed experiments. S. Kovats conceived of the study, designed experiments, analyzed data and wrote the manuscript.

Competing interests

The authors declare no competing interests.

Corresponding author

Correspondence to Susan Kovats.

Supplementary information

About this article

Publication history