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In Vitro Assay of Plasmodium-Infected Red Blood Cell Killing by Cytotoxic Lymphocytes
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In Vitro Assay of Plasmodium-Infected Red Blood Cell Killing by Cytotoxic Lymphocytes

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Activation of human NK cells by Plasmodium-infected red blood cells

Amir Horowitz1, Eleanor M Riley

  • 1Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, UK.

Methods in Molecular Biology (Clifton, N.J.)
|September 20, 2012
PubMed

Insights

This protocol details how to measure human Natural Killer (NK) cell activation after exposure to malaria-infected red blood cells using flow cytometry. It outlines methods for parasite culture, cell separation, and analysis of CD69 and IFN-γ expression.

Area of Science:

  • Immunology
  • Infectious Diseases
  • Parasitology

Background:

  • Malaria poses a significant global health challenge, necessitating research into host immune responses.
  • Human Natural Killer (NK) cells play a crucial role in the innate immune response to infections, including malaria.
  • Understanding NK cell activation is vital for developing effective malaria control strategies.

Purpose of the Study:

  • To describe a detailed protocol for assessing human NK cell activation in vitro.
  • To provide a standardized method for evaluating NK cell responses to Plasmodium falciparum-infected red blood cells.
  • To facilitate research on NK cell-mediated immunity against malaria.

Main Methods:

  • In vitro propagation and purification of Plasmodium falciparum parasites.
  • Separation of peripheral blood mononuclear cells (PBMC) from human blood via density centrifugation.
  • Flow cytometry analysis of NK cell surface CD69 expression and intracellular IFN-γ accumulation following in vitro stimulation.

Main Results:

  • The protocol enables the assessment of NK cell activation markers (CD69 and IFN-γ) upon stimulation with malaria-infected erythrocytes.
  • Typical flow cytometry (FACS) plots demonstrating NK cell activation are presented.
  • The described methods allow for quantitative analysis of NK cell responses.

Conclusions:

  • This protocol provides a reproducible method for studying human NK cell activation in the context of malaria.
  • The findings contribute to a better understanding of the innate immune response to Plasmodium falciparum.
  • This methodology can be applied to investigate immunomodulatory therapies for malaria.

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