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Flow cytometric analysis of macaque whole blood for antigen‐specific intracellular cytokine production by T lymphocytes
Author(s) -
Keeney Talitha S.,
Nomura Laurel E.,
Maecker Holder T.,
Jagannadha Sastry K.
Publication year - 2003
Publication title -
journal of medical primatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.31
H-Index - 42
eISSN - 1600-0684
pISSN - 0047-2565
DOI - 10.1034/j.1600-0684.2003.00005.x
Subject(s) - antigen , immunology , cytokine , whole blood , macaque , cd8 , biology , interferon gamma , immune system , tumor necrosis factor alpha , flow cytometry , t lymphocyte , paleontology
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‐ γ ) and tumor necrosis factor alpha (TNF‐ α ) in CD4 + as well as CD8 + T lymphocytes. We observed significantly higher levels of TNF‐ α compared with IFN‐ γ 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‐ α 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.