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Updated: Jun 18, 2026
Generation of Human Alloantigen-specific T Cells from Peripheral Blood
Published on: November 21, 2014
Optimization and validation of a robust human T-cell culture method for monitoring phenotypic and polyfunctional
Yun Lin1, Humilidad F Gallardo, Geoffrey Y Ku
1Ludwig Center for Cancer Immunotherapy, Immunology Program, Sloan-Kettering Institute, Newy York, New York 10021, USA.
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)-1beta, 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-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.
