Effect of HLA-G on proliferation in peripheral blood lymphocytes of childbearing age women and Treg cell subsets
10.3969/j.issn.1000-484X.2015.05.025
- VernacularTitle: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
- Author:
Juan ZHANG
;
Xiaoyun LIU
;
Fumin LIU
;
Xia FENG
- Publication Type:Journal Article
- Keywords:
HLA-G;
TNF-α;
Treg;
Cell proliferation
- From:
Chinese Journal of Immunology
2015;(5):690-694
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the effect of HLA-Gon proliferation in peripheral blood lymphocytes of childbearing age women and Treg cell subsets,and investigate the mechanisms of immune tolerance in pregnancy.Methods:The high expression of HLA-G cho-riocarcinoma cell lines JEG-3 cells with peripheral blood lymphocytes( PBLC) of healthy childbearing age women co-culture,using the HLA-G neutralizing antibodies(87G)and recombinant human tumor necrosis factor receptor typeⅡ-antibody fusion protein(rhTNFR:Fc)to intervene.Experiments were divided into seven groups:①JEG-3+PBLC culture group;②JEG-3+PBLC+87G culture group;③JEG-3+PBLC non-contact culture group;④JEG-3+PBLC+87G non-contact culture group;⑤The control group(PBLC group);⑥JEG-3+PBLC+rhTNFR:Fc culture group;⑦JEG-3+PBLC+87G+rhTNFR:Fc culture group.Detected the PBLC proliferation inhibition by CCK-8 method and the expression of TNF-αmRNA by RT-PCR in①-⑤groups.The proportion of Treg cells were detected by flow cytometry in①-⑦groups.Results:The assay of CCK-8 showed that the PBLC proliferation inhibition rate of 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±5.56)%,(14.67±4.04)%,(37.67±2.31)% and(8.33±3.21)%,there was a statistically significant difference on each group ( P<0.05 ).The result of RT-PCR showed that the expression of TNF-αmRNA in JEG-3+PBLC culture group and JEG-3+PBLC non-contact culture group were significantly lower than the control group.Compared with the corresponding non-87G group,the expression of TNF-αmRNA increased significantly after the intervention of 87G,P<0.05.Compared with the control group, the proportion of Treg cells of JEG-3+PBLC culture group detected by flow cytometry was significantly increased(P<0.05).There was no significant difference between JEG-3+PBLC non-contact culture group and control group ( P>0.05 ).Compared with the JEG-3+PBLC culture group,the proportion of Treg cells of JEG-3+PBLC+87G culture group was significantly decreased(P<0.05).Compared with JEG-3+PBLC culture group,the proportion of Treg cells of JEG-3+PBLC+rhTNFR:Fc culture group was significantly increased( P<0.05).Set JEG-3+PBLC+rhTNFR:Fc culture group as control,the proportion of Treg cells of JEG-3+PBLC+rh TNF:FC+87G culture was significantly decreased,but obviously higher than JEG-3+PBLC+87G culture group,there was a statistically significant difference on each group(P<0.05).Conclusion:HLA-G can inhibit peripheral blood lymphocyte proliferation of childbearing age women and inhibit the expression of TNF-α,and up-regulate the proportion of Treg cells.
