Flow cytometric analysis of macaque whole blood for antigen-specific intracellular cytokine production by T lymphocytes |
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Authors: | Keeney Talitha S Nomura Laurel E Maecker Holder T Sastry K Jagannadha |
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Institution: | Department of Veterinary Sciences, The University of Texas M. D. Anderson Cancer Center, Bastrop, TX 78602, USA. |
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Abstract: | We report here the standardized conditions for stimulation of macaque whole blood samples with various protein or peptide antigens, and production of significant intracellular levels of interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) in CD4+ as well as CD8+ T lymphocytes. We observed significantly higher levels of TNF-alpha compared with IFN-gamma in both CD4+ and CD8+ T lymphocytes from all the macaque whole blood samples stimulated with staphylococcal enterotoxin B (SEB) as an antigen. Similarly, when whole blood samples from rhesus macaques immunized with an HIV envelope peptide cocktail vaccine were stimulated with either the peptide cocktail or recombinant gp160, we observed production of significant levels of TNF-alpha by both CD4+ and CD8+ T lymphocytes. These results strongly support the utility of the whole blood cytokine flow cytometry methodology for determining antigen-specific immune responses of macaques in vaccine studies. |
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Keywords: | CD4 CD8 flow cytometry IFN-γ IL-2 Macaca mulatta non-human primate rhesus macaque SEB TNF-α |
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