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目的研究人脐静脉内皮细胞系(ECV304)表达的HLA-G1对同种异体外周血单个核细胞(PBMC)Th1/Th2型细胞因子分泌的影响。方法采用脂质体介导的基因转染技术将pcDNA3-HLA-G1转入ECV304,以间接免疫荧光技术在蛋白质水平上检测HLA-G1分子在ECV304上的表达;以表达HLA-G1的ECV304作为刺激细胞,灭活后,与健康人PBMC共同培养,用ELISA检测上清液中Th1/Th2型细胞因子的浓度,观察HLA-G1对同种异体抗原激活的PBMC分泌Th1/Th2型细胞因子的影响。结果与转染pcDNA3空质粒的对照组相比,HLA-G1能使PBMC的IL-10分泌增加(P<0.05),而IL-2,IFN-γI、L-4分泌无明显影响。结论HLA-G1能引起由同种异体抗原激活的PBMC分泌IL-10增加,提示HLA-G1有可能使Th1/Th2型细胞因子向Th2型偏移。
Objective To investigate the effect of HLA-G1 expressed by human umbilical vein endothelial cell line (ECV304) on the secretion of Th1 / Th2 cytokines in allogeneic peripheral blood mononuclear cells (PBMCs). Methods The pcDNA3-HLA-G1 was transfected into ECV304 by liposome-mediated gene transfection and the expression of HLA-G1 on ECV304 was detected by indirect immunofluorescence. The ECV304 expressing HLA-G1 was used as The cells were stimulated, inactivated and cultured with PBMCs of healthy people. The concentrations of Th1 / Th2 cytokines in the supernatants were detected by ELISA. The effect of HLA-G1 on the secretion of Th1 / Th2 cytokines by alloantigen-activated PBMCs influences. Results Compared with the control group transfected with pcDNA3 empty plasmid, HLA-G1 increased the secretion of IL-10 in PBMCs (P <0.05), but had no effect on the secretion of IL-2, IFN-γI and L-4. Conclusion HLA-G1 can induce the increase of IL-10 secretion by PBMCs activated by alloantigen, suggesting that HLA-G1 might shift Th1 / Th2 cytokines to Th2 type.