Multicolor flow cytometry analyses of cellular immune response in rhesus macaques

Hong He, Amy N. Courtney, Eric Wieder, K. Jagannadha Sastry

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

The rhesus macaque model is currently the best available model for HIV-AIDS with respect to understanding the pathogenesis as well as for the development of vaccines and therapeutics. Here, we describe a method for the detailed phenotypic and functional analyses of cellular immune responses, specifically intracellular cytokine production by CD4+ and CD8+ T cells as well as the individual memory subsets. We obtained precise quantitative and qualitative measures for the production of interferon gamma (INF-) and interleukin (IL) -2 in both CD4+ and CD8+ T cells from the rhesus macaque PBMC stimulated with PMA plus ionomycin (PMA+I). The cytokine profiles were different in the different subsets of memory cells. Furthermore, this protocol provided us the sensitivity to demonstrate even minor fractions of antigen specific CD4+ and CD8+ T cell subsets within the PBMC samples from rhesus macaques immunized with an HIV envelope peptide cocktail vaccine developed in our laboratory. The multicolor flow cytometry technique is a powerful tool to precisely identify different populations of T cells with cytokine-producing capability following non-specific or antigen-specific stimulation.

Original languageEnglish (US)
Article numbere1743
JournalJournal of Visualized Experiments
Issue number38
DOIs
StatePublished - Apr 2010

Keywords

  • Cytokine production
  • FACS
  • Flow cytometry
  • HIV
  • Immune response
  • Intracellular cytokine staining
  • Issue 38
  • Jove immunology
  • Rhesus macaque
  • T cells

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

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