【摘要】 目的:研究肝癌特异性识别基因TCRVβ7.1表达重组体转染外周血单个核细胞(PBMCs)后对T细胞细胞因子表达的影响和信号通路的激活作用。方法:以肝癌特异性T细胞受体Vβ7.1转染PBMCs,流式细胞仪检测TCRVβ7.1表达量;Western blotting检测ERK1/2表达量及磷酸化水平(p-ERK)的改变;用ELISA法检测白细胞介素-4(IL-4)、γ-干扰素(IFN-γ)的表达量。结果:TCRVβ7.1基因转染健康人PBMCs并得到有效表达,转染TCRVβ7.1基因的PBMCs与肝癌细胞共培养后ERK蛋白磷酸化水平明显高于未转染组(P<0.01)。p-ERK1/2水平与T细胞激活有关。ELISA结果表明,转染PBMCs细胞与肝癌细胞共培养后,IFN-γ水平明显高于未转染组,而IL-4无明显改变。结论:TCRVβ7.1转染PBMCs与肝癌细胞共培养后,ERK信号通路被激活,IFN-γ表达增高。更多还原

【Abstract】 AIM:To investigate the effect of tumor-specific T cell receptor(TCR) gene transfection on production of cytokine and signaling activation in T cells.METHODS:TCRVβ7.1 gene was transferred into peripheral blood mononuclear cells(PBMCs) obtained from healthy adults,and the expression of Vβ7.1 was detected by flow cytometry before and after transfection.The total quantities of protein and phosphorylation of ERK1/2 were detected by Western blotting.The expressions of IL-4 and IFN-γ were detected by ELISA.RESULTS:The results of flow cytometry showed that TCRVβ7.1 protein was efficiently expressed after transfection.The phosphorylation level of ERK increased significantly in TCRVβ7.1-modified PBMCs,and was related with the activation of T cells.The expression of IFN-γ was significantly higher in TCR-transfected cells than that in non-transfected cells.The expression of IL-4,however,has no distinct difference between groups.CONCLUSION:The transfection of TCRVβ7.1 induces phosphorylation of ERK1/2 and production of IFN-γ,and activates T lymphocytes.更多还原