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Natural killer cells migrate into and control simian immunodeficiency virus replication in lymph node follicles in African green monkeys

Nature Medicine volume 23, pages 12771286 (2017) | Download Citation

Abstract

Natural killer (NK) cells play an essential role in antiviral immunity, but knowledge of their function in secondary lymphoid organs is incomplete. Lymph node follicles constitute a major viral reservoir during infections with HIV-1 and simian immunodeficiency virus of macaques (SIVmac). In contrast, during nonpathogenic infection with SIV from African green monkeys (SIVagm), follicles remain generally virus free. We show that NK cells in secondary lymphoid organs from chronically SIVagm-infected African green monkeys (AGMs) were frequently CXCR5+ and entered and persisted in lymph node follicles throughout the follow-up (240 d post-infection). These follicles were strongly positive for IL-15, which was primarily presented in its membrane-bound form by follicular dendritic cells. NK cell depletion through treatment with anti-IL-15 monoclonal antibody during chronic SIVagm infection resulted in high viral replication rates in follicles and the T cell zone and increased viral DNA in lymph nodes. Our data suggest that, in nonpathogenic SIV infection, NK cells migrate into follicles and play a major role in viral reservoir control in lymph nodes.

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References

  1. 1.

    , , & Immune responses during spontaneous control of HIV and AIDS: what is the hope for a cure? Phil. Trans. R. Soc. Lond. B 369, 20130436 (2014).

  2. 2.

    & The lymph node in HIV pathogenesis. Semin. Immunol. 20, 187–195 (2008).

  3. 3.

    & HIV-associated chronic immune activation. Immunol. Rev. 254, 78–101 (2013).

  4. 4.

    et al. Early stages of simian immunodeficiency virus infection in lymph nodes. Evidence for high viral load and successive populations of target cells. Am. J. Pathol. 144, 1226–1237 (1994).

  5. 5.

    et al. Expansion of HIV-specific T follicular helper cells in chronic HIV infection. J. Clin. Invest. 122, 3271–3280 (2012).

  6. 6.

    et al. Differential infection patterns of CD4+ T cells and lymphoid tissue viral burden distinguish progressive and nonprogressive lentiviral infections. Blood 120, 4172–4181 (2012).

  7. 7.

    et al. CD4 T follicular helper cell dynamics during SIV infection. J. Clin. Invest. 122, 3281–3294 (2012).

  8. 8.

    et al. Persistent simian immunodeficiency virus infection drives differentiation, aberrant accumulation, and latent infection of germinal center follicular T helper cells. J. Virol. 90, 1578–1587 (2015).

  9. 9.

    et al. Decreased T follicular regulatory cell/T follicular helper cell (TFH) in simian immunodeficiency virus–infected rhesus macaques may contribute to accumulation of TFH in chronic infection. J. Immunol. 195, 3237–3247 (2015).

  10. 10.

    Pathobiology of HIV/SIV-associated changes in secondary lymphoid tissues. Immunol. Rev. 254, 65–77 (2013).

  11. 11.

    & Follicular dendritic cells initiate and maintain infection of the germinal centers by human immunodeficiency virus. Curr. Top. Microbiol. Immunol. 201, 141–159 (1995).

  12. 12.

    et al. Lymphoid germinal centers are reservoirs of human immunodeficiency virus type 1 RNA. J. Infect. Dis. 164, 1051–1057 (1991).

  13. 13.

    et al. B cell follicle sanctuary permits persistent productive simian immunodeficiency virus infection in elite controllers. Nat. Med. 21, 132–139 (2015).

  14. 14.

    et al. CXCR5+ follicular cytotoxic T cells control viral infection in B cell follicles. Nat. Immunol. 17, 1187–1196 (2016).

  15. 15.

    et al. PD-1+ and follicular helper T cells are responsible for persistent HIV-1 transcription in treated aviremic individuals. Nat. Med. 22, 754–761 (2016).

  16. 16.

    & TFH in HIV latency and as sources of replication-competent virus. Trends Microbiol. 24, 338–344 (2016).

  17. 17.

    , , , & Functions of natural killer cells. Nat. Immunol. 9, 503–510 (2008).

  18. 18.

    & Innate immune control of HIV. Cold Spring Harb. Perspect. Med. 2, a007070 (2012).

  19. 19.

    & Natural killer cell distribution and trafficking in human tissues. Front. Immunol. 3, 347 (2012).

  20. 20.

    et al. Natural killer cell behavior in lymph nodes revealed by static and real-time imaging. J. Exp. Med. 203, 619–631 (2006).

  21. 21.

    , , & Natural killer cells actively patrol peripheral lymph nodes forming stable conjugates to eliminate MHC-mismatched targets. Proc. Natl. Acad. Sci. USA 104, 12081–12086 (2007).

  22. 22.

    et al. Dynamic behavior of NK cells during activation in lymph nodes. Blood 114, 3227–3234 (2009).

  23. 23.

    et al. In vivo imaging of inflammasome activation reveals a subcapsular macrophage burst response that mobilizes innate and adaptive immunity. Nat. Med. 22, 64–71 (2016).

  24. 24.

    et al. Early viral replication in lymph nodes provides HIV with a means by which to escape NK-cell-mediated control. Eur. J. Immunol. 41, 2729–2740 (2011).

  25. 25.

    , , , & Innate immune cell responses in non pathogenic versus pathogenic SIV infections. Curr. Opin. Virol. 19, 37–44 (2016).

  26. 26.

    et al. Acute loss of intestinal CD4+ T cells is not predictive of simian immunodeficiency virus virulence. J. Immunol. 179, 3035–3046 (2007).

  27. 27.

    et al. Lack of dichotomy between virus load of peripheral blood and lymph nodes during long-term simian immunodeficiency virus infection of African green monkeys. Virology 219, 367–375 (1996).

  28. 28.

    et al. High levels of viral replication during primary simian immunodeficiency virus SIVagm infection are rapidly and strongly controlled in African green monkeys. J. Virol. 74, 7538–7547 (2000).

  29. 29.

    et al. Viral load in tissues during the early and chronic phase of non-pathogenic SIVagm infection. J. Med. Primatol. 33, 83–97 (2004).

  30. 30.

    et al. Wide range of viral load in healthy African green monkeys naturally infected with simian immunodeficiency virus. J. Virol. 74, 11744–11753 (2000).

  31. 31.

    et al. Simian immunodeficiency virus replicates to high levels in naturally infected African green monkeys without inducing immunologic or neurologic disease. J. Virol. 75, 2262–2275 (2001).

  32. 32.

    et al. Early divergence in lymphoid tissue apoptosis between pathogenic and nonpathogenic simian immunodeficiency virus infections of nonhuman primates. J. Virol. 82, 1175–1184 (2008).

  33. 33.

    & SIVagm: genetic and biological features associated with replication. Front. Biosci. 8, d1170–d1185 (2003).

  34. 34.

    et al. Acute SIV infection in sooty mangabey monkeys is characterized by rapid virus clearance from lymph nodes and absence of productive infection in germinal centers. PLoS One 8, e57785 (2013).

  35. 35.

    et al. Innate immune responses and rapid control of inflammation in African green monkeys treated or not with interferon-α during primary SIVagm infection. PLoS Pathog. 10, e1004241 (2014).

  36. 36.

    et al. Effect of anti-IL-15 administration on T cell and NK cell homeostasis in rhesus macaques. J. Immunol. 197, 1183–1198 (2016).

  37. 37.

    et al. In vivo evidence for a dependence on interleukin 15 for survival of natural killer cells. Blood 100, 3633–3638 (2002).

  38. 38.

    et al. Differential effects of IL-15 on the generation, maintenance and cytotoxic potential of adaptive cellular responses induced by DNA vaccination. Vaccine 33, 1188–1196 (2015).

  39. 39.

    , , , & Accumulation of cytotoxic CD16+ NK cells in simian immunodeficiency virus–infected lymph nodes associated with in situ differentiation and functional anergy. J. Virol. 89, 6887–6894 (2015).

  40. 40.

    et al. A B-cell-homing chemokine made in lymphoid follicles activates Burkitt's lymphoma receptor-1. Nature 391, 799–803 (1998).

  41. 41.

    , & CXCR5+ T cells: follicular homing takes center stage in T-helper-cell responses. Trends Immunol. 23, 250–254 (2002).

  42. 42.

    et al. Generation of cellular immune memory and B-cell immunity is impaired by natural killer cells. Nat. Commun. 6, 6375 (2015).

  43. 43.

    & Homeostasis of the antibody response: immunoregulation by NK cells. Science 222, 581–585 (1983).

  44. 44.

    , , & Human B cell activation by autologous NK cells is regulated by CD40–CD40 ligand interaction: role of memory B cells and CD5+ B cells. J. Immunol. 167, 6132–6139 (2001).

  45. 45.

    , & Interactions between B lymphocytes and NK cells. FASEB J. 8, 1012–1018 (1994).

  46. 46.

    et al. IL-15 promotes activation and expansion of CD8+ T cells in HIV-1 infection. J. Clin. Invest. 126, 2745–2756 (2016).

  47. 47.

    , , , & NK cells are primed by ANRS MVAHIV-infected DCs, via a mechanism involving NKG2D and membrane-bound IL-15, to control HIV-1 infection in CD4+ T cells. Eur. J. Immunol. 44, 2370–2379 (2014).

  48. 48.

    , , & Follicular dendritic cells produce IL-15 that enhances germinal center B cell proliferation in membrane-bound form. J. Immunol. 173, 6676–6683 (2004).

  49. 49.

    et al. A human CD34+ subset resides in lymph nodes and differentiates into CD56bright natural killer cells. Immunity 22, 295–304 (2005).

  50. 50.

    & Trans-presentation: a novel mechanism regulating IL-15 delivery and responses. Immunol. Lett. 127, 85–92 (2010).

  51. 51.

    et al. IL-15.IL-15Rα complex shedding following trans-presentation is essential for the survival of IL-15 responding NK and T cells. Proc. Natl. Acad. Sci. USA 111, 8565–8570 (2014).

  52. 52.

    , & IL-15-mediated induction of LFA-1 is a late step required for cytotoxic differentiation of human NK cells from CD34+Lin bone marrow cells. J. Immunol. 171, 683–690 (2003).

  53. 53.

    , , & Dendritic cells support the in vivo development and maintenance of NK cells via IL-15 trans-presentation. J. Immunol. 183, 4948–4956 (2009).

  54. 54.

    & Expansion or depletion of T follicular helper cells during HIV infection: consequences for B cell responses. Curr. HIV Res. 11, 595–600 (2013).

  55. 55.

    , & Follicular dendritic cells in HIV-induced lymphadenopathy and AIDS. APMIS Suppl. 8, 16–23 (1989).

  56. 56.

    et al. Simian immunodeficiency virus (SIV)-specific CD8+ T-cell responses in vervet African green monkeys chronically infected with SIVagm. J. Virol. 82, 11577–11588 (2008).

  57. 57.

    et al. Gag p27-specific B- and T-cell responses in simian immunodeficiency virus SIVagm-infected African green monkeys. J. Virol. 83, 2770–2777 (2009).

  58. 58.

    et al. Inhibition of adaptive immune responses leads to a fatal clinical outcome in SIV-infected pigtailed macaques but not vervet African green monkeys. PLoS Pathog. 5, e1000691 (2009).

  59. 59.

    et al. Experimental depletion of CD8+ cells in acutely SIVagm-infected African green monkeys results in increased viral replication. Retrovirology 7, 42 (2010).

  60. 60.

    et al. Nonpathogenic SIV infection of African green monkeys induces a strong but rapidly controlled type I IFN response. J. Clin. Invest. 119, 3544–3555 (2009).

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Acknowledgements

This work was supported by the Investissements d'Avenir program managed by the National Agency of Research (ANR) under reference ANR-10-LABX-77, the ANRS and the L'Oréal Foundation. The Infectious Disease Models and Innovative Therapies (IDMIT) center in Fontenay-aux-Roses, France, was funded by the French government's Investissements d'Avenir program for infrastructures (PIA) under grant ANR-11-INBS-0008, the PIA grant ANR-10-EQPX-02-01 funding the FlowCyTech facility at IDMIT. R.K.R. was supported by National Institutes of Health (NIH) grant RO1 DE026014. The anti-IL-15 monoclonal antibody was a gift from the NIH Nonhuman Primate Reagent Resource, supported by AI126683 and OD010976. N.H. and M.J.P. were recipients of postdoctoral fellowships from the French Vaccine Research Institute (Créteil, France) and Sidaction, respectively. P.R. and T.G.-T. received a PhD fellowship from the University Paris Diderot, Sorbonne Paris Cité (BIOSPC), and the Pasteur-Paris University PhD program supported by the Institut Carnot Pasteur Microbes et Santé, respectively. We are grateful to the veterinarians and the staff of the IDMIT animal facility, in particular V. Contreras, B. Delache, J.-M. Helies, V. Monnet, J. Morin and C. Joubert, for their excellent work. We thank L. Irbah, T. Kortulewski and C. Chapon for access to the stellar IDMIT imaging core facility as well as C. Cassan, S. Guenounou and A. Cosma for access to the state-of-the-art IDMIT FlowCyTech core facility.

Author information

Affiliations

  1. Unit on HIV, Inflammation and Persistence, Institut Pasteur, Paris, France.

    • Nicolas Huot
    • , Beatrice Jacquelin
    • , Thalia Garcia-Tellez
    • , Philippe Rascle
    • , Mickaël J Ploquin
    •  & Michaela Müller-Trutwin
  2. Vaccine Research Institute, Créteil, France.

    • Nicolas Huot
    • , Philippe Rascle
    •  & Michaela Müller-Trutwin
  3. Université Paris Diderot, Sorbonne Paris Cité, Paris, France.

    • Philippe Rascle
  4. Epidemiology Unit, Institut Pasteur, Paris, France.

    • Yoann Madec
  5. Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.

    • R Keith Reeves
  6. Commissariat à l'Energie Atomique et aux Énergies Alternatives (CEA), Université Paris Sud 11, INSERM U1184, Immunology of Viral Infections and Autoimmune Diseases (IMVA), Institut de Biologie François Jacob (IBJF), Infectious Disease Models and Innovative Therapies (IDMIT) Department, Fontenay-aux-Roses, France.

    • Nathalie Derreudre-Bosquet

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Contributions

N.H. contributed to the project design, set up methods and performed the experiments; N.D.-B. provided samples; N.H., B.J., T.G.-T., P.R. and M.J.P. processed the samples; P.R. prepared the IL15 probe; N.H. and Y.M. performed the statistical analyses; N.H., B.J., R.K.R. and M.M.-T. analyzed and interpreted the data; B.J. prepared the ethical protocols; N.H. and M.M.-T. wrote the manuscript; and M.M.-T. conceived the project and directed the research.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Michaela Müller-Trutwin.

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DOI

https://doi.org/10.1038/nm.4421