摘要
目的探讨肝癌特异性T细胞受体基因转染对记忆性T细胞体外诱导分化的影响。方法以肝癌特异性T细胞受体Vβ7.1转染PBMC,流式细胞术检测目的基因表达。肝癌细胞BEL-7402刺激诱导记忆性T细胞;以流式细胞术检测记忆性T细胞标志性表面分子表达验证记忆性T细胞表型及分化亚群;以AnnexinV-PI双染检测肿瘤细胞的凋亡,ELISA检测细胞因子IFN-γ分泌探讨记忆性T细胞免疫功能。结果TCRVβ7.1基因成功转染到T细胞并得到有效表达。与PBMC对照组相比,TCRVβ7.1转染组CD45RO表达显著上升。以CD45RA及CCR7表达分析记忆性T细胞亚群显示其表型主要为效应型记忆性T细胞。分化后的记忆性T细胞诱导肿瘤细胞凋亡率及IFN-γ分泌显著上升。结论肿瘤特异性TCR基因转染可促进以效应型记忆性T细胞(TEM)为主的记忆性T细胞分化,TEM将通过诱导凋亡与分泌细胞因子作用发挥其免疫功能。
Objective To investigate effect of tumor-specific T cell receptor gene transfection on memory T cell differentiation in vitro. Methods TCRVβ7.1 gene was transferred into peripheral blood mononuclear cells （PBMCs） obtained from healthy adults, and the expression of V137.1 was detected by flow cytometry before and after the transfection. Memory T cell differentiation was induced by stimulation with the hepatocarcinoma cell line BEL-7402 in vitro. The expression of surface molecules CD45RO, CD45RA and CCR7 was analyzed by flow cytometry to identify the phenotype and subsets of the memory T cells. Fluorescence-activated cell sorting was performed to detect the apoptosis of the tumor cells, and enzyme-linked immunoabsorbent assay was used to determine the production of interferon-γ（IFN-γ） for assessing the immune function of the memory T cells. Results Flow cytometry showed that TCRVβ7.1 gene was efficiently expressed after transfection. After stimulation by the tumor cells in vitro, the expression of CD45RO in TCRVβ7.1 gene-modified T cells increased gradually, and analysis of the coexpression of CD45RA and CCR7 revealed that the effector memory T cells constituted the majority of the differentiated memory T cells. The apoptotic rate of the tumor cells induced by the T cells increased significantly with also obviously increased INF-γ secretion in the memory T cells. Conclusion Tumor-specific TCRVβ7.1 gene transfection can promote the differentiation of the memory T cells, the majority of which belongs to effector memory T cells that perform immune functions by inducing apoptosis and cytokine secretion.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2008年第3期313-316,共4页
Journal of Southern Medical University
基金
国家自然科学基金(30572124)
广东省自然科学基金(034046)
广东省科技计划项目(2004B31201001)~~
关键词
肿瘤
T细胞受体
转染
记忆性T细胞
分化
tumor
T cell receptor
transfection
memory T cells
differentiation
