Journal
CYTOTHERAPY
Volume 11, Issue 7, Pages 912-U127Publisher
ELSEVIER SCI LTD
DOI: 10.3109/14653240903136987
Keywords
Antigen-specific T cell; CD4 T cell; CD8 T cell; immune monitoring; phenotype; polyfunctional analysis
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Funding
- Swim Across America
- Experimental Therapeutics Center of MSKCC
- Ludwig Foundation
- Damon Runyon-Lilly Clinical Investigator Award
- NATIONAL CANCER INSTITUTE [P01CA033049] Funding Source: NIH RePORTER
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Background aims Monitoring cellular immune responses is one prerequisite for the rational development of cancer vaccines. Methods We 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)-gamma, interleukin (IL)-2, macrophage inflammatory protein (MIP)-1 beta, tumor necrosis factor (TNF)-alpha and CD107a. Results Autologous 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-specifi c CD4(+) and CD8(+) T cells were detectable using this method. Conclusions Our 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.
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