Detection of antigen-specific T cell cytokine expression in whole blood by flow cytometry.
We have recently described a highly sensitive flow cytometric technique, based on the ability to detect single cell expression of cytokines, to simultaneously quantitate and phenotypically characterize antigen-specific memory/effector T cells in PBMC cultures. In this report, we describe a simplified procedural modification which enables the rapid detection of low frequency memory CD4+ and CD8+ T cells expressing cytokines in response to soluble antigen in whole blood. When compared with T cell responses in PBMC cultures, whole blood cultures demonstrated similar but slightly higher percentages of T cells responsive to specific antigen. In addition, T cell responses to cytomegalovirus in whole blood were observed only in sensitized (seropositive) individuals, and CD4+ T cell responses could be blocked by anti-class II MHC antibodies. This procedure may provide a means to examine direct effects of pharmacological drug concentrations on T cell immunity in clinical samples.
Becton Dickinson Immunocytometry Systems, San Jose, CA 95131, USA. maria_suni@bdis.com
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D000956:Antigens, Viral / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D002478:Cells, Cultured; D016207:Cytokines / Q000097:blood*; D003586:Cytomegalovirus Infections / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007156:Immunologic Memory; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
D000956:Antigens, Viral; D016207:Cytokines