Abstract
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)-γ, interleukin (IL)-2, macrophage inflammatory protein (MIP)-1β, tumor necrosis factor (TNF)-α 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-specific 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.
Acknowledgments
We thank Dr M. Roederer for providing the SPICE software, Dr Immanuel Luescher from the Tetramer Core, Lausanne Branch, Ludwig Institute of Cancer Research, for providing all the tetramers used in these experiments, Dr Bo Dupont and Alice Yeh from his laboratory for performing HLA analysis on one patient, and Dr. Miguel Perales, who provided editorial advice. This work was supported by Swim Across America, the Experimental Therapeutics Center of MSKCC and Ludwig Foundation. J. D. Wolchok was supported by a Damon Runyon-Lilly Clinical Investigator Award.
Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.