Flow cytometric analysis of macaque whole blood for antigen-specific intracellular cytokine production by T lymphocytes.
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.
Department of Veterinary Sciences, The University of Texas M. D. Anderson Cancer Center, Bastrop, TX 78602, USA.
Talitha S
TS
Keeney
Laurel E
LE
Nomura
Holder T
HT
Maecker
K Jagannadha
KJ
Sastry
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000502:physiology; D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects / Q000502:physiology; D004768:Enterotoxins / Q000494:pharmacology; D005434:Flow Cytometry; D019302:HIV Envelope Protein gp160 / Q000494:pharmacology; D007371:Interferon-gamma / Q000096:biosynthesis*; D008253:Macaca mulatta / Q000097:blood* / Q000276:immunology; D011994:Recombinant Proteins / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D004768:Enterotoxins; D019302:HIV Envelope Protein gp160; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; C031818:enterotoxin B, staphylococcal; D007371:Interferon-gamma