Skip to main content

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

  • Protocol
  • First Online:
Natural Killer Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 612))

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 heparinised 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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 279.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Newman, K. C., and Riley, E. M. (2007) Whatever turns you on: accessory-cell-dependent activation of NK cells by pathogens. Nat Rev Immunol 7, 279–291.

    Article  CAS  PubMed  Google Scholar 

  2. Haller, D., Serrant, P., Granato, D., Schiffrin, E. J., and Blum, S. (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.

    CAS  PubMed  Google Scholar 

  3. Newman, K. C., Korbel, D. S., Hafalla, J. C., and Riley, E. M. (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 

  4. Gorak, P. M., Engwerda, C. R., and Kaye, P. M. (1998) Dendritic cells, but not macrophages, produce IL-12 immediately following Leishmania donovani infection. Eur J Immunol 28, 687–695.

    Article  CAS  PubMed  Google Scholar 

  5. Dalod, M., Hamilton, T., Salomon, R., Salazar-Mather, T. P., Henry, S. C., Hamilton, J. D., and Biron, C. A. (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  CAS  PubMed  Google Scholar 

  6. Artavanis-Tsakonas, K., Eleme, K., McQueen, K. L., Cheng, N. W., Parham, P., Davis, D. M., and Riley, E. M. (2003) Activation of a subset of human NK cells upon contact with Plasmodium falciparum-infected erythrocytes. J Immunol 171, 5396–5405.

    CAS  PubMed  Google Scholar 

  7. Guma, M., Cabrera, C., Erkizia, I., Bofill, M., Clotet, B., Ruiz, L., and Lopez-Botet, M. (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 

  8. LaBonte, M. L., McKay, P. F., and Letvin, N. L. (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  CAS  PubMed  Google Scholar 

  9. Korbel, D. S., Newman, K. C., Almeida, C. R., Davis, D. M., and Riley, E. M. (2005) Heterogeneous human NK cell responses to Plasmodium falciparum-infected erythrocytes. J Immunol 175, 7466–7473.

    CAS  PubMed  Google Scholar 

  10. Mohan, K., Moulin, P., and Stevenson, M. M. (1997) Natural killer cell cytokine production, not cytotoxicity, contributes to resistance against blood-stage Plasmodium chabaudi AS infection. J Immunol 159, 4990–4998.

    CAS  PubMed  Google Scholar 

  11. Choudhury, H. R., Sheikh, N. A., Bancroft, G. J., Katz, D. R., and De Souza, J. B. (2000) Early nonspecific immune responses and immunity to blood-stage nonlethal Plasmodium yoelii malaria. Infect Immun 68, 6127–6132.

    Article  CAS  PubMed  Google Scholar 

  12. De Souza, J. B., Williamson, K. H., Otani, T., and Playfair, J. H. (1997) Early gamma interferon responses in lethal and nonlethal murine blood-stage malaria. Infect Immun 65, 1593–1598.

    PubMed  Google Scholar 

  13. Stevenson, M. M., and Riley, E. M. (2004) Innate immunity to malaria. Nat Rev Immunol 4, 169–180.

    Article  CAS  PubMed  Google Scholar 

  14. Kim, C. C., Parikh, S., Sun, J. C., Myrick, A., Lanier, L. L., Rosenthal, P. J., and DeRisi, J. L. (2008) Experimental malaria infection triggers early expansion of natural killer cells. Infect Immun 76, 5873–5882.

    Article  CAS  PubMed  Google Scholar 

  15. Artavanis-Tsakonas, K., and Riley, E. M. (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.

    CAS  PubMed  Google Scholar 

  16. Baratin, M., Roetynck, S., Lepolard, C., Falk, C., Sawadogo, S., Uematsu, S., Akira, S., Ryffel, B., Tiraby, J. G., Alexopoulou, L., Kirschning, C. J., Gysin, J., Vivier, E., and Ugolini, S. (2005) Natural killer cell and macrophage cooperation in MyD88-dependent innate responses to Plasmodium falciparum. Proc Natl Acad Sci U S A 102, 14747–14752.

    Article  CAS  PubMed  Google Scholar 

  17. Maghazachi, A. A. (2003) G protein-coupled receptors in natural killer cells. J Leukoc Biol 74, 16–24.

    Article  CAS  PubMed  Google Scholar 

  18. Fehniger, T. A., Cooper, M. A., Nuovo, G. J., Cella, M., Facchetti, F., Colonna, M., and Caligiuri, M. A. (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  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Horowitz, A., Riley, E.M. (2010). Activation of Human NK Cells by Malaria-Infected Red Blood Cells. In: Campbell, K. (eds) Natural Killer Cell Protocols. Methods in Molecular Biology, vol 612. Humana Press. https://doi.org/10.1007/978-1-60761-362-6_29

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-362-6_29

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-361-9

  • Online ISBN: 978-1-60761-362-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics