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.