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Cytotoxic CD8+ T cells recognize and kill Plasmodium vivax–infected reticulocytes

Nature Medicine volume 24, pages 1330–1336 (2018)Cite this article

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Abstract

Plasmodium vivax causes approximately 100 million clinical malaria cases yearly1,2. The basis of protective immunity is poorly understood and thought to be mediated by antibodies3,4. Cytotoxic CD8+ T cells protect against other intracellular parasites by detecting parasite peptides presented by human leukocyte antigen class I on host cells. Cytotoxic CD8+ T cells kill parasite-infected mammalian cells and intracellular parasites by releasing their cytotoxic granules5,6. Perforin delivers the antimicrobial peptide granulysin and death-inducing granzymes into the host cell, and granulysin then delivers granzymes into the parasite. Cytotoxic CD8+ T cells were thought to have no role against Plasmodium spp. blood stages because red blood cells generally do not express human leukocyte antigen class I7. However, P. vivax infects reticulocytes that retain the protein translation machinery. Here we show that P. vivax–infected reticulocytes express human leukocyte antigen class I. Infected patient circulating CD8+ T cells highly express cytotoxic proteins and recognize and form immunological synapses with P. vivax–infected reticulocytes in a human leukocyte antigen–dependent manner, releasing their cytotoxic granules to kill both host cell and intracellular parasite, preventing reinvasion. P. vivax–infected reticulocytes and parasite killing is perforin independent, but depends on granulysin, which generally efficiently forms pores only in microbial membranes8. We find that P. vivax depletes cholesterol from the P. vivax–infected reticulocyte cell membrane, rendering it granulysin-susceptible. This unexpected T cell defense might be mobilized to improve P. vivax vaccine efficacy.

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Fig. 1: Increased frequency of activated CD8+ T cells in the peripheral blood of P. vivax patients.
Fig. 2: Increased HLA-ABC in P. vivax–infected reticulocytes.
Fig. 3: CD8+ T cells are activated by and lyse autologous P. vivax–infected reticulocytes.
Fig. 4: Granulysin binds to infected reticulocytes and mediates host cell lysis and parasite killing.

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Acknowledgements

We thank Dr. Kasturi Haldar and Kenneth Rock for scientific discussions and suggestions during the development of this work. We are grateful to the Program for Technological Development in Tools for Health–PDTIS-FIOCRUZ for use of its facilities, and to the clinic, laboratory and administrative staff, as well as field workers and subjects from Porto Velho who participated in the study. This study was funded by the National Institutes of Health (1R01NS098747 to R.T.G.; the Amazonian-ICEMR U19 AI089681 to C.J., L.R.A., and R.T.G.; 1R01AI116577 and R21AI131632-01 to R.T.G. and J.L.); National Institute of Science and Technology for Vaccines/Conselho Nacional de Desenvolvimento Científico e Tecnológico (465293/2014-0 to C.J., L.R.A., D.B.P. and R.T.G.) and Fundação de Amparo à Pesquisa do Estado de Minas Gerais (RED-00012-14 and APQ-00653-16 to C.J. and R.T.G.); and Fundação de Amparo à Pesquisa do Estado de São Paulo (2016/23618-8 to R.T.G.). C.J., L.R.A., A.T.-C., and R.T.G. are recipients of CNPq fellowships; P.A.C.C. and C.R.R.B. are fellows from FAPEMIG; and C.J. is a fellow from Coordenação de Aperfeiçoamento de Pessoal de Ensino Superior.

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Authors and Affiliations

  1. Instituto René Rachou, Fundação Oswaldo Cruz, Belo Horizonte, Brazil

    Caroline Junqueira, Camila R. R. Barbosa, Pedro A. C. Costa, Andréa Teixeira-Carvalho, Guilherme Castro & Lis R. Antonelli

  2. Program in Cellular and Molecular Medicine, Boston Children’s Hospital, Boston, MA, USA

    Caroline Junqueira, Sumit Sen Santara, Rafael P. Barbosa, Farokh Dotiwala & Judy Lieberman

  3. Department of Pediatrics, Harvard Medical School, Boston, MA, USA

    Caroline Junqueira, Sumit Sen Santara, Rafael P. Barbosa, Farokh Dotiwala & Judy Lieberman

  4. Centro de Pesquisas em Medicina Tropical, Porto Velho, Brazil

    Dhelio B. Pereira

  5. Plataforma de Medicina Translacional, Fundação Oswaldo Cruz, Ribeirão Preto, Brazil

    Ricardo T. Gazzinelli

  6. Departamento de Bioquímica e Imunologia, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Ricardo T. Gazzinelli

  7. Division of Infectious Disease and Immunology, University of Massachusetts Medical School, Worcester, MA, USA

    Ricardo T. Gazzinelli

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Contributions

C.J., J.L., and R.T.G. conceived the study. C.J. and R.T.G. supervised this study. D.B.P. recruited the patients and healthy donors; C.J., L.R.A., A.T.-C., J.L., and R.T.G. designed and analyzed experiments. C.J., C.R.R.B., L.R.A., P.A.C.C., A.T.-C., G.C., and R.P.B. performed experiments. S.S.S. and F.D. contributed reagents. C.J., C.R.R.B., P.A.C.C., and L.R.A. prepared figures and helped with manuscript preparation. C.J., J.L., and R.T.G. wrote the paper.

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Correspondence to Caroline Junqueira, Judy Lieberman or Ricardo T. Gazzinelli.

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Junqueira, C., Barbosa, C.R.R., Costa, P.A.C. et al. Cytotoxic CD8+ T cells recognize and kill Plasmodium vivax–infected reticulocytes. Nat Med 24, 1330–1336 (2018). https://doi.org/10.1038/s41591-018-0117-4

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