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 language | English (US) |
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Article number | e1743 |
Journal | Journal of Visualized Experiments |
Issue number | 38 |
DOIs | |
State | Published - 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
- General Neuroscience
- General Chemical Engineering
- General Biochemistry, Genetics and Molecular Biology
- General Immunology and Microbiology