HLA-G对育龄女性外周血淋巴细胞增殖及Treg细胞亚群的影响

发布时间：2019-06-16 18:19

【摘要】：目的:研究HLA-G对育龄女性外周血中淋巴细胞(PBLC)增殖及Treg细胞亚群的影响,探讨其在妊娠免疫耐受中的作用机制。方法:高表达HLA-G的绒癌细胞系JEG-3细胞与健康育龄女性外周血淋巴细胞(PBLC)分组共培养,利用HLA-G中和抗体(87G)和重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rh TNFR∶Fc)进行干预,实验分为7组:1JEG-3+PBLC培养组;2JEG-3+PBLC+87G培养组;3JEG-3+PBLC非接触培养组;4JEG-3+PBLC+87G非接触培养组;5对照组(单纯PBLC组);6JEG-3+PBLC+rh TNFR:Fc培养组;7JEG-3+PBLC+87G+rh TNFR:Fc培养组。1~5每组CCK-8法检测各组PBLC增殖抑制情况,RT-PCR方法检测TNF-αmRNA的表达,1~7每组流式细胞术检测Treg细胞的比例。结果:CCK-8法显示,JEG-3+PBLC培养组、JEG-3+PBLC+87G培养组、JEG-3+PBLC非接触培养组、JEG-3+PBLC+87G非接触培养组PBLC增殖抑制率分别为(48.00±5.56)%、(14.67±4.04)%、(37.67±2.31)%、(8.33±3.21)%,各组间比较差异均有统计学意义,P值均0.05;RT-PCR结果显示JEG-3+PBLC培养组、JEG-3+PBLC非接触培养组TNF-αmRNA的表达量明显低于对照组,加入87G干预后,TNF-αmRNA的表达量较相应无87G组明显上升,P值均0.05。流式细胞检测显示,与对照组相比,JEG-3+PBLC培养组Treg细胞比例明显上升(P0.05),而JEG-3+PBLC非接触培养组差异无统计学意义(P0.05),JEG-3+PBLC+87G培养组Treg细胞比例较JEG-3+PBLC培养组明显下降(P0.05);JEG-3+PBLC+rh TNFR:Fc培养组与JEG-3+PBLC培养组相比,Treg细胞比例明显上升(P0.05);以JEG-3+PBLC+rh TNFR:Fc培养组为对照,JEG-3+PBLC+rh TNF:FC+87G培养组Treg细胞比例明显下降,但明显高于JEG-3+PBLC+87G培养组,各组间比较差异均有统计学意义,P值均0.05。结论:HLA-G可以抑制育龄女性外周血淋巴细胞的增殖及抑制TNF-α的表达,上调Treg细胞的比例。
[Abstract]:Aim: to study the effect of HLA-G on (PBLC) proliferation and Treg cell subsets in peripheral blood of women of childbearing age, and to explore the mechanism of Treg cell subsets in pregnancy immune tolerance. Methods: chorionic carcinoma cell line JEG-3 cells with high expression of HLA-G were co-cultured with (PBLC) of healthy female peripheral blood lymphocytes of childbearing age. HLA-G neutralizing antibody (87G) and recombinant human type 2 tumor necrosis factor receptor-antibody fusion protein (rh TNFR:Fc) were intervened. the cells were divided into 7 groups: 1JEG-3 PBLC culture group, 2JEG-3 PBLC 87G culture group, 3JEG-3 PBLC non-contact culture group, and 2JEG-3 PBLC 87G culture group, 2JEG-3 PBLC 87G culture group, 3JEG-3 PBLC non-contact culture group, 2JEG-3 PBLC 87G culture group, 3JEG-3 PBLC non-contact culture group. 4JEG-3 PBLC 87G non-contact culture group; 5 control group (simple PBLC group); 6JEG-3 PBLC rh TNFR:Fc culture group; 7JEG-3 PBLC 87G rh TNFR:Fc culture group. 1 CCK-8 assay was used to detect the proliferation inhibition of PBLC in each group, RT-PCR method was used to detect the expression of TNF- 伪 mRNA, and flow cytometry was used to detect the proportion of Treg cells in each group. Results: CCK-8 assay showed that the inhibition rates of PBLC proliferation in JEG-3 PBLC culture group, JEG-3 PBLC 87G culture group, JEG-3 PBLC non-contact culture group and JEG-3 PBLC 87G non-contact culture group were (48.00 卤5.56)%, (14.67 卤4.04)%, (37.67 卤2.31)% and (8.33 卤3.21)%, respectively. The results of RT-PCR showed that the expression of TNF- 伪 mRNA in JEG-3 PBLC culture group and JEG-3 PBLC non-contact culture group was significantly lower than that in control group. The expression of TNF- 伪 mRNA in JEG-3 PBLC non-contact culture group was significantly higher than that in non-87G group (P = 0.05). Flow cytometry showed that the proportion of Treg cells in JEG-3 PBLC culture group was significantly higher than that in control group (P 0.05), but there was no significant difference in JEG-3 PBLC non-contact culture group (P 0.05). The proportion of Treg cells in JEG-3 PBLC 87G culture group was significantly lower than that in JEG-3 PBLC culture group (P 0.05), and the proportion of Treg cells in JEG-3 PBLC rh TNFR:Fc culture group was significantly higher than that in JEG-3 PBLC culture group (P 0.05). Compared with JEG-3 PBLC rh TNFR:Fc culture group, the proportion of Treg cells in JEG-3 PBLC rh TNF:FC 87G culture group decreased significantly, but it was significantly higher than that in JEG-3 PBLC 87G culture group. Conclusion: HLA-G can inhibit the proliferation of peripheral blood lymphocytes and the expression of TNF- 伪 in women of childbearing age, and up-regulate the proportion of Treg cells.
【作者单位】：徐州医学院研究生学院;
【分类号】：R714

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