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通过构建人趋化因子CCL25基因过表达慢病毒载体并转染人原代胸腺上皮细胞(TEC),探讨过表达CCL25对人TEC黏附分子表达的影响,为研究CCL25在重症肌无力患者胸腺异常表达的作用奠定基础。PCR和DNA测序鉴定携带CCL25基因的慢病毒构建成功,病毒滴度约为4×108 Tu/ml,慢病毒对TEC感染效率达80%。Western blotting结果显示感染组细胞CCL25蛋白表达量显著增高。RT-PCR结果显示,与对照组相比,感染组HLA-A、HLA-DR表达无显著变化,P-selectin、VCAM-1和ICAM-1表达显著增高,提示我们人胸腺上皮细胞CCL25基因的过表达可一定程度影响TEC的功能特性,可能以通过增加P-selectin、VCAM-1和ICAM-1的表达量来影响淋巴祖细胞的胸腺归巢或胸腺内胸腺细胞的阳性选择过程,进而引起胸腺细胞发育异常,细胞亚群比例失调。
To investigate the effect of CCL25 on the expression of human TEC adhesion molecules by over-expressing the lentiviral vector CCL25 and transfection of human primary thymus epithelial cells (TEC). To investigate the expression of CCL25 in patients with myasthenia gravis The role of foundation. PCR and DNA sequencing identified that the CCL25 gene-carrying lentivirus was successfully constructed with a virus titer of about 4 × 108 Tu / ml and a lentiviral efficiency of TEC of 80%. Western blotting results showed that the expression of CCL25 protein in infected cells was significantly increased. The results of RT-PCR showed that there was no significant change in the expression of HLA-A and HLA-DR, but the expression of P-selectin, VCAM-1 and ICAM-1 were significantly increased in the infected group as compared with the control group, suggesting that the human CCL25 gene in human thymic epithelial cells Overexpression may affect the functional properties of TEC to a certain extent and may affect the positive selection of thymus lymphocytes or thymus lymphocytes in lymphoid progenitor cells by increasing the expression of P-selectin, VCAM-1 and ICAM-1, Thymocyte dysplasia, imbalance of cell subsets.