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Malaria pp 447–464Cite as

Activation of Human NK Cells by Plasmodium-Infected Red Blood Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 923))

Abstract

This chapter describes a protocol to assess activation of human NK cells following in vitro stimulation with malaria-infected red blood cells. Activation is assessed by flow cytometry, staining for cell surface expression of CD69 and accumulation of intracellular IFN-γ. Procedures are described for in vitro propagation and purification of Plasmodium falciparum parasites, separation of peripheral blood mononuclear cells from heparinized blood by density centrifugation, in vitro culture of PBMC and for staining and analysis of PBMC by flow cytometry. Some examples of typical FACS plots are shown.

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References

  1. Ferreira A et al (1986) Inhibition of development of exoerythrocytic forms of malaria ­parasites by gamma-interferon. Science 232:881–884

    Article  PubMed  CAS  Google Scholar 

  2. Romero P et al (1989) Cloned cytotoxic T cells recognize an epitope in the circumsporozoite protein and protect against malaria. Nature 341:323–326

    Article  PubMed  CAS  Google Scholar 

  3. Ing R et al (2006) Interaction of mouse dendritic cells and malaria-infected erythrocytes: uptake, maturation, and antigen presentation. J Immunol 176:441–450

    PubMed  CAS  Google Scholar 

  4. McGilvray ID et al (2000) Nonopsonic monocyte/macrophage phagocytosis of Plasmodium falciparum-parasitized erythrocytes: a role for CD36 in malarial clearance. Blood 96:3231–3240

    PubMed  CAS  Google Scholar 

  5. Mohan K et al (1997) Natural killer cell cytokine production, not cytotoxicity, contributes to resistance against blood-stage Plasmodium ­chabaudi AS infection. J Immunol 159:4990–4998

    PubMed  CAS  Google Scholar 

  6. Choudhury HR et al (2000) Early nonspecific immune responses and immunity to blood-stage nonlethal Plasmodium yoelii malaria. Infect Immun 68:6127–6132

    Article  PubMed  CAS  Google Scholar 

  7. De Souza JB et al (1997) Early gamma interferon responses in lethal and nonlethal murine blood-stage malaria. Infect Immun 65:1593–1598

    PubMed  Google Scholar 

  8. Stevenson MM, Riley EM (2004) Innate immunity to malaria. Nat Rev Immunol 4:169–180

    Article  PubMed  CAS  Google Scholar 

  9. Roland J et al (2006) NK cell responses to Plasmodium infection and control of intrahepatic parasite development. J Immunol 177:1229–1239

    PubMed  CAS  Google Scholar 

  10. Hansen DS et al (2007) NK cells stimulate recruitment of CXCR3+ T cells to the brain during Plasmodium berghei-mediated cerebral malaria. J Immunol 178:5779–5788

    PubMed  CAS  Google Scholar 

  11. Kim CC et al (2008) Experimental malaria infection triggers early expansion of natural killer cells. Infect Immun 76:5873–5882

    Article  PubMed  CAS  Google Scholar 

  12. Artavanis-Tsakonas K, Riley EM (2002) Innate immune response to malaria: rapid induction of IFN-gamma from human NK cells by live Plasmodium falciparum-infected erythrocytes. J Immunol 169:2956–2963

    PubMed  CAS  Google Scholar 

  13. Korbel DS et al (2005) Heterogeneous human NK cell responses to Plasmodium falciparum-infected erythrocytes. J Immunol 175:7466–7473

    PubMed  CAS  Google Scholar 

  14. Baratin M et al (2005) Natural killer cell and macrophage cooperation in MyD88-dependent innate responses to Plasmodium falciparum. Proc Natl Acad Sci USA 102:14747–14752

    Article  PubMed  CAS  Google Scholar 

  15. Newman KC, Riley EM (2007) Whatever turns you on: accessory-cell-dependent activation of NK cells by pathogens. Nat Rev Immunol 7:279–291

    Article  PubMed  CAS  Google Scholar 

  16. Artavanis-Tsakonas K et al (2003) Activation of a subset of human NK cells upon contact with Plasmodium falciparum-infected erythrocytes. J Immunol 171:5396–5405

    PubMed  CAS  Google Scholar 

  17. Newman KC et al (2006) Cross-talk with myeloid accessory cells regulates human natural killer cell interferon-gamma responses to malaria. PLoS Pathog 2:e118

    Article  PubMed  Google Scholar 

  18. Horowitz A et al (2010) Cross-talk between T cells and NK cells generates rapid effector responses to Plasmodium falciparum-infected erythrocytes. J Immunol 184:6043–6052

    Article  PubMed  CAS  Google Scholar 

  19. Korbel DS et al (2009) Killer Ig-like receptor (KIR) genotype predicts the capacity of human KIR-positive CD56dim NK cells to respond to pathogen-associated signals. J Immunol 182:6426–6434

    Article  PubMed  CAS  Google Scholar 

  20. Haller D et al (2002) Activation of human NK cells by staphylococci and lactobacilli requires cell contact-dependent costimulation by autologous monocytes. Clin Diagn Lab Immunol 9:649–657

    PubMed  CAS  Google Scholar 

  21. Gorak PM et al (1998) Dendritic cells, but not macrophages, produce IL-12 immediately following Leishmania donovani infection. Eur J Immunol 28:687–695

    Article  PubMed  CAS  Google Scholar 

  22. Dalod M et al (2003) Dendritic cell responses to early murine cytomegalovirus infection: subset functional specialization and differential regulation by interferon alpha/beta. J Exp Med 197:885–898

    Article  PubMed  CAS  Google Scholar 

  23. Maghazachi AA (2003) G protein-coupled receptors in natural killer cells. J Leukoc Biol 74:16–24

    Article  PubMed  CAS  Google Scholar 

  24. Fehniger TA et al (2003) CD56bright natural killer cells are present in human lymph nodes and are activated by T cell-derived IL-2: a potential new link between adaptive and innate immunity. Blood 101:3052–3057

    Article  PubMed  CAS  Google Scholar 

  25. Guma M et al (2006) Human cytomegalovirus infection is associated with increased proportions of NK cells that express the CD94/NKG2C receptor in aviremic HIV-1-positive patients. J Infect Dis 194:38–41

    Article  PubMed  Google Scholar 

  26. LaBonte ML et al (2006) Evidence of NK cell dysfunction in SIV-infected rhesus monkeys: impairment of cytokine secretion and NKG2C/C2 expression. Eur J Immunol 36:2424–2433

    Article  PubMed  CAS  Google Scholar 

  27. Miyagami T, Waki S (1985) In vitro cultivation of Plasmodium falciparum under aerobic atmosphere in a CO2 incubator. J Parasitol 71:262–263

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Eleanor M. Riley .

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Horowitz, A., Riley, E.M. (2012). Activation of Human NK Cells by Plasmodium-Infected Red Blood Cells. In: Ménard, R. (eds) Malaria. Methods in Molecular Biology, vol 923. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-026-7_31

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  • DOI: https://doi.org/10.1007/978-1-62703-026-7_31

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-025-0

  • Online ISBN: 978-1-62703-026-7

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