Optimization and validation of a robust human T-cell culture method for monitoring phenotypic and polyfunctional antigen-specific CD4 and CD8 T-cell responses

Background aimsMonitoring cellular immune responses is one prerequisite for the rational development of cancer vaccines.MethodsWe describe an extensive effort to optimize and validate quantitatively an in vitro T-cell culture method by determining the phenotype and function of both CD4+ and CD8+ T cells, including measurement of the phenotype markers CCR7, CD45RA, CD28 and CD27 and the functional markers interferon (IFN)-γ, interleukin (IL)-2, macrophage inflammatory protein (MIP)-1β, tumor necrosis factor (TNF)-α and CD107a.ResultsAutologous peripheral blood mononuclear cells (PBMC) were potent stimulators that expanded antigen (Ag)-specific CD8+ T cells during short-term culture with the addition of IL-2 and IL-15 cytokines. Polyfunctional Ag-specific CD4+ and CD8+ T cells were detectable using this method.ConclusionsOur culture system represents a robust human T-cell culture protocol that permits phenotypic, quantitative and qualitative evaluation of vaccine-induced CD4+ and CD8+ T-cell responses.