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。
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 transfecfion. 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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2009年第8期1505-1508,共4页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30572124)
广东省自然科学基金资助项目(No.034046)
广东省科技计划资助项目(No.2004B31201001)
