人类胎盘enEVTs通过NCAM1募集白细胞并抑制Th1细胞分化  

Human placental enEVTs recruit leukocytes through NCAM1 and inhibit Th1 cell differentiation

作　　者：马烨灵 张巧英[1] 郑长武[1] 冉娜 方丹娜 MA Ye-ling;ZHANG Qiao-ying;ZHENG Chang-wu;RAN Na;FANG Dan-na(Medical School,Shaoxing University,Shaoxing 312000,China.)

机构地区：[1]绍兴文理学院医学院,浙江绍兴312000

出　　处：《中国病理生理杂志》2022年第9期1634-1644,共11页Chinese Journal of Pathophysiology

基　　金：绍兴文理学院科研启动基金(No.20210022)。

摘　　要：目的:探讨血管内绒毛外滋养层细胞(endovascular extravillous trophoblasts,enEVTs)调控妊娠免疫耐受的机制。方法:通过磁珠分选技术获得人工流产早孕蜕膜中的原代enEVTs和间质绒毛外滋养层细胞(interstitial extravillous trophoblasts,iEVTs),再将enEVTs和iEVTs培养上清分别处理Naive CD4^(+)T细胞,并将enEVTs培养上清处理组定义为实验组,iEVTs培养上清处理组定义为对照组。通过流式细胞术比较实验组和对照组诱导CD4^(+)IFNγ^(+)1型辅助性T细胞(type 1 T helper cells,Th1细胞)分化比例;通过免疫荧光实验比较enEVTs和iEVTs中神经细胞黏附分子1(neural cell adhesion molecule 1,NCAM1)的表达情况;通过enEVTs和iEVTs分别与白细胞共培养,比较enEVTs对白细胞的黏附作用;进一步对37例人类胎盘进行绒毛间隙的苏木精-伊红(hematoxylin-eosin,HE)染色,进行白细胞募集的统计和检测。结果:流式细胞术结果显示,在细胞体外共培养实验中enEVTs相比于iEVTs显著抑制CD4^(+)IFNγ^(+)Th1细胞的分化(P<0.05);免疫荧光结果表明,enEVTs相比于iEVTs特异高表达NCAM1;原代细胞共培养结果证明,enEVTs可黏附母血中的白细胞;HE染色结果显示,enEVTs可富集母血白细胞。结论:(1)enEVTs特异表达NCAM1并募集母血的白细胞于enEVTs周围;(2)enEVTs抑制Th1细胞的分化,进而促进妊娠过程中的免疫耐受。AIM:Study the mechanism of endovascular extravillous trophoblasts(enEVTs)in regulating immune tolerance during pregnancy.METHODS:Interstitial extravillous trophoblasts(iEVTs),enEVTs and na?ve CD4^(+)T were obtained from the decidua of early pregnancy by magnetic bead sorting. The naive CD4^(+)T cells were treated with enEVTs or iEVTs supernatant. The enEVTs supernatant-treated group was defined as the experimental group,and the iEVTs supernatant-treated group was defined as the control group. The proportion of differentiated CD4^(+)IFNγ^(+)type 1 T helper cells(Th1 cells)was analyzed by flow cytometry analysis. The expression of neural cell adhesion molecule 1(NCAM1)in iEVTs and enEVTs was analyzed. The adhesion of enEVTs to leukocytes was studied by co-culture experiment. Furthermore,in order to detect the recruitment of leukocytes,hematoxylin-eosin(HE)staining of the intervillous space was performed in 37 human placentas.RESULTS:Flow cytometry analysis revealed that enEVTs significantly inhibited the differentiation of CD4^(+)IFNγ^(+)Th1 cells compared with iEVTs. Immunofluorescence results showed that NCAM1 was highly expressed in enEVTs. Primary cell co-culture results showed that enEVTs could adhere to maternal leukocytes. The results of HE staining further revealed that maternal leukocytes were recruited by enEVTs.CONCLUSION:Human placental enEVTs recruit leukocytes through NCAM1. The enEVTs inhibit the differentiation of Th1 cells and promote immune tolerance during pregnancy.

关 键 词：血管内绒毛外滋养层细胞 神经细胞黏附分子1 TH1细胞 白细胞 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学] R363.21[医药卫生—基础医学]