Ludwig Center for Cancer Immunotherapy, Immunology Program, Sloan-Kettering Institute, Newy York, New York 10021, USA.
Cytotherapy. 2009;11(7):912-22. doi: 10.3109/14653240903136987.
Monitoring cellular immune responses is one prerequisite for the rational development of cancer vaccines.
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.
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.
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.
监测细胞免疫反应是癌症疫苗合理开发的前提条件之一。
我们描述了一项优化和验证体外 T 细胞培养方法的努力，通过确定 CD4(+)和 CD8(+)T 细胞的表型和功能来实现，包括测量表型标志物 CCR7、CD45RA、CD28 和 CD27 以及功能标志物干扰素 (IFN)-γ、白细胞介素 (IL)-2、巨噬细胞炎症蛋白 (MIP)-1β、肿瘤坏死因子 (TNF)-α 和 CD107a。
自体外周血单核细胞 (PBMC) 是有效的刺激物，在添加白细胞介素 (IL)-2 和 IL-15 细胞因子的短期培养中可扩增抗原 (Ag)-特异性 CD8(+)T 细胞。使用这种方法可以检测到多能 Ag-特异性 CD4(+)和 CD8(+)T 细胞。
我们的培养系统代表了一种强大的人类 T 细胞培养方案，可允许对疫苗诱导的 CD4(+)和 CD8(+)T 细胞反应进行表型、定量和定性评估。
