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CD8+ T lymphocytes protective against malaria liver stages are primed in skin-draining lymph nodes

Nature Medicine volume 13, pages 10351041 (2007) | Download Citation

Abstract

The success of immunization with irradiated sporozoites is unparalleled among the current vaccination approaches against malaria, but its mechanistic underpinnings have yet to be fully elucidated. Using a model mimicking natural infection by Plasmodium yoelii, we delineated early events governing the development of protective CD8+ T-cell responses to the circumsporozoite protein. We demonstrate that dendritic cells in cutaneous lymph nodes prime the first cohort of CD8+ T cells after an infectious mosquito bite. Ablation of these lymphoid sites greatly impairs subsequent development of protective immunity. Activated CD8+ T cells then travel to systemic sites, including the liver, in a sphingosine-1-phosphate (S1P)-dependent fashion. These effector cells, however, no longer require bone marrow–derived antigen-presenting cells for protection; instead, they recognize antigen on parenchymal cells—presumably parasitized hepatocytes. Therefore, we report an unexpected dichotomy in the tissue restriction of host responses during the development and execution of protective immunity to Plasmodium.

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References

  1. 1.

    , , & Visualization of peptide-specific T-cell immunity and peripheral tolerance induction in vivo. Immunity 1, 327–339 (1994).

  2. 2.

    & Delayed migration of Plasmodium sporozoites from the mosquito bite site to the blood. Am. J. Trop. Med. Hyg. 57, 426–429 (1997).

  3. 3.

    & Intravital microscopy demonstrating antibody-mediated immobilisation of Plasmodium berghei sporozoites injected into skin by mosquitoes. Int. J. Parasitol. 34, 991–996 (2004).

  4. 4.

    et al. Quantitative imaging of Plasmodium transmission from mosquito to mammal. Nat. Med. 12, 220–224 (2006).

  5. 5.

    , , & Protective immunity produced by the injection of x-irradiated sporozoites of Plasmodium berghei. Nature 216, 160–162 (1967).

  6. 6.

    , , & Specificity of protection of man immunized against sporozoite-induced falciparum malaria. Am. J. Med. Sci. 266, 398–403 (1973).

  7. 7.

    et al. Gamma interferon, CD8+ T cells and antibodies required for immunity to malaria sporozoites. Nature 330, 664–666 (1987).

  8. 8.

    , , , & CD8+ T cells (cytotoxic/suppressors) are required for protection in mice immunized with malaria sporozoites. Proc. Natl. Acad. Sci. USA 85, 573–576 (1988).

  9. 9.

    , & The relative contribution of antibodies, CD4+ and CD8+ T cells to sporozoite-induced protection against malaria. Immunology 80, 1–5 (1993).

  10. 10.

    , , , & Single immunizing dose of recombinant adenovirus efficiently induces CD8+ T-cell–mediated protective immunity against malaria. J. Immunol. 158, 1268–1274 (1997).

  11. 11.

    et al. The circumsporozoite protein is an immunodominant protective antigen in irradiated sporozoites. Nature 444, 937–940 (2006).

  12. 12.

    et al. Swift development of protective effector functions in naive CD8+ T cells against malaria liver stages. J. Exp. Med. 194, 173–180 (2001).

  13. 13.

    , , , & Short-term antigen presentation and single clonal burst limit the magnitude of the CD8+ T-cell responses to malaria liver stages. Proc. Natl. Acad. Sci. USA 99, 11819–11824 (2002).

  14. 14.

    et al. Early self-regulatory mechanisms control the magnitude of CD8+ T cell responses against liver stages of murine malaria. J. Immunol. 171, 964–970 (2003).

  15. 15.

    et al. In vivo depletion of CD11c+ dendritic cells abrogates priming of CD8+ T cells by exogenous cell-associated antigens. Immunity 17, 211–220 (2002).

  16. 16.

    , , & Parasitology. Malaria vaccines: back to the future? Science 307, 528–530 (2005).

  17. 17.

    et al. Intravital observation of Plasmodium berghei sporozoite infection of the liver. PLoS Biol. 3, e192 (2005).

  18. 18.

    , & Apoptotic Plasmodium-infected hepatocytes provide antigens to liver dendritic cells. J. Infect. Dis. 191, 1576–1581 (2005).

  19. 19.

    & Quantitative dynamics of Plasmodium yoelli sporozoite transmission by infected anopheline mosquitoes. Infect. Immun. 73, 4363–4369 (2005).

  20. 20.

    , , & Plasmodium sporozoites trickle out of the injection site. Cell. Microbiol. 9, 1215–1222 (2007).

  21. 21.

    et al. The role of intrahepatic lymphocytes in mediating protective immunity induced by attenuated Plasmodium berghei sporozoites. Immunol. Rev. 174, 123–134 (2000).

  22. 22.

    , , , & IL-4 receptor expression on CD8+ T cells is required for the development of protective memory responses against liver stages of malaria parasites. J. Exp. Med. 202, 551–560 (2005).

  23. 23.

    , & B7-DC crosslinking restores antigen uptake and augments antigen-presenting cell function by matured dendritic cells. Proc. Natl. Acad. Sci. USA 102, 11438–11443 (2005).

  24. 24.

    et al. Systemic activation of dendritic cells by Toll–like receptor ligands or malaria infection impairs cross-presentation and antiviral immunity. Nat. Immunol. 7, 165–172 (2006).

  25. 25.

    et al. FTY720 immunosuppression impairs effector T cell peripheral homing without affecting induction, expansion, and memory. J. Immunol. 164, 5761–5770 (2000).

  26. 26.

    , & FTY720: sphingosine-1-phosphate receptor 1 in the control of lymphocyte egress and endothelial barrier function. Am. J. Transplant. 4, 1019–1025 (2004).

  27. 27.

    & Phenotype and function of rat dendritic cell subsets. APMIS 111, 756–765 (2003).

  28. 28.

    , & Collection of lymph-borne dendritic cells in the rat. Nat. Protoc. 1, 2263–2270 (2006).

  29. 29.

    et al. A T-cell clone directed at the circumsporozoite protein which protects mice against both Plasmodium yoelii and Plasmodium berghei. J. Immunol. 149, 2103–2109 (1992).

  30. 30.

    et al. Interferon-gamma-independent CD8+ T-cell-mediated protective anti-malaria immunity elicited by recombinant adenovirus. Parasite Immunol. 22, 157–160 (2000).

  31. 31.

    , , & Immunoelectron microscopic localization of circumsporozoite antigen in the differentiating exoerythrocytic trophozoite of Plasmodium berghei. Cell Biol. Int. Rep. 12, 123–129 (1988).

  32. 32.

    , & Antigen presentation by liver cells controls intrahepatic T-cell trapping, whereas bone marrow–derived cells preferentially promote intrahepatic T-cell apoptosis. J. Immunol. 167, 667–673 (2001).

  33. 33.

    et al. Cellular and molecular mechanisms of liver tolerance. Immunol. Rev. 213, 101–118 (2006).

  34. 34.

    & The dissection of CD8+ T cells during liver-stage infection. Curr. Top. Microbiol. Immunol. 297, 1–24 (2005).

  35. 35.

    & Malaria sporozoites release circumsporozoite protein from their apical end and translocate it along their surface. J. Protozool. 38, 411–421 (1991).

  36. 36.

    et al. Migration of Plasmodium sporozoites through cells before infection. Science 291, 141–144 (2001).

  37. 37.

    et al. Priming of CD8+ T cell responses following immunization with heat-killed Plasmodium sporozoites. Eur. J. Immunol. 36, 1179–1186 (2006).

  38. 38.

    , , & Kupffer cell engraftment across the major histocompatibility barrier in mice: bone marrow origin, class II antigen expression, and antigen-presenting capacity. J. Pediatr. Gastroenterol. Nutr. 11, 525–533 (1990).

  39. 39.

    et al. Commitment of bone marrow cells to hepatic stellate cells in mouse. J. Hepatol. 40, 255–260 (2004).

  40. 40.

    , , & Nomadic or sessile: can Kupffer cells function as portals for malaria sporozoites to the liver? Cell Microbiol. 8, 1537–1546 (2006).

  41. 41.

    et al. Kupffer cells are obligatory for Plasmodium yoelii sporozoite infection of the liver. Cell. Microbiol. 9, 397–412 (2007).

  42. 42.

    et al. Reevaluation of bone marrow–derived cells as a source for hepatocyte regeneration. Cell Transplant. 13, 659–666 (2004).

  43. 43.

    et al. CD8+ cytolytic T-cell clones derived against the Plasmodium yoelii circumsporozoite protein protect against malaria. Int. Immunol. 3, 579–585 (1991).

  44. 44.

    et al. Priming with recombinant influenza virus followed by administration of recombinant vaccinia virus induces CD8+ T-cell-mediated protective immunity against malaria. Proc. Natl. Acad. Sci. USA 90, 5214–5218 (1993).

  45. 45.

    , , & Preferential localization of effector memory cells in nonlymphoid tissue. Science 291, 2413–2417 (2001).

  46. 46.

    et al. The dendritic cell populations of mouse lymph nodes. J. Immunol. 167, 741–748 (2001).

  47. 47.

    , & ELISPOT assay to measure antigen-specific murine CD8+ T cell responses. J. Immunol. Methods 252, 207–218 (2001).

  48. 48.

    et al. Detection of malaria liver-stages in mice infected through the bite of a single Anopheles mosquito using a highly sensitive real-time PCR. Int. J. Parasitol. 31, 1499–1502 (2001).

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Acknowledgements

The authors wish to thank G. Milon for exciting discussions, and D. Griffin, A.F. Scott, N.J. Singh and W.R. Heath for helpful comments and suggestions. I. Villa performed a few preliminary experiments. Flow cytometry was done in the Becton Dickinson Immune Function Laboratory at the Johns Hopkins Bloomberg School of Public Health. This work was supported by US National Institutes of Health grant no. AI44375. S.K. was supported by a joint fellowship from Turkish Scientific Technical Research Council and the North Atlantic Treaty Organization (TUBITAK-NATO).

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Affiliations

  1. Department of Molecular Microbiology and Immunology, Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, 615 N. Wolfe Street, Baltimore, Maryland 21205, USA.

    • Sumana Chakravarty
    • , Ian A Cockburn
    • , Salih Kuk
    • , Michael G Overstreet
    •  & Fidel Zavala
  2. Department of Microbiology and Immunology, University of Maryland School of Medicine, 660 West Redwood St., Baltimore, Maryland 21201, USA.

    • John B Sacci

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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Fidel Zavala.

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DOI

https://doi.org/10.1038/nm1628

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