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血管内皮细胞系表达的HLA-G1抑制同种T细胞增殖及活性 预览 被引量:1

Immunotolerance-inducing effect on allogeneic T cells by HLA-G1 expressed on ECV304
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摘要 目的研究血管内皮细胞系人脐静脉内皮细胞(ECV304)表达的HLA-G1分子对同种异体T细胞增殖的作用.方法采用脂质体介导的DNA转染技术,将构建的真核表达质粒pcDNA3-HLA-G1转染ECV304,用免疫荧光法检测其表达的HLA-G1分子.将表达HLA-G1的ECV304与同种异体T细胞进行混合细胞培养后,用四甲基偶氮唑(MTT)比色法检测ECV304表达HLA-G1对外周血T细胞增殖和抗原特异性细胞毒T淋巴细胞(CTL)杀伤活性的影响.结果ECV304转染pcDNA3-HLA-G1后可稳定表达HLA-G1分子;转染空质粒pcDNA3和转染pcDNA3-HLA-G1的T细胞的刺激指数分别为(1.59±0.41)和(1.33±0.46),二组比较,差异有统计学意义(P<0.05);细胞毒实验显示,抗原特异性CTL对转染空质粒pcDNA3和转染pcDNA3-HLA-G1的ECV304的杀伤率分别为(64.81±3.07)%和(52.33±4.48)%,二组比较,差异有统计学意义(P<0.05).结论HLA-G1分子可以抑制同种异体T细胞的增殖和CTL的细胞毒作用. Objective To study the inhibitory effects of HLA-G1 expressed in ECV304 on the proliferation of allogeneic T cells. Methods The recombinant plasmid pcDNA3-HLA-G1 which contained a full-length cDNA of HLA-G1 was constructed, and the ECV304 cell was transfected with the pcDNA3-HLA-G1 by using the lipofectin transfection. The expressed HLA-G1 on the ceil surface was checked by specific monoclonal antibody (G11 E5) with indirect immunofluorescence assay and FCM. The HLA-G1 expressed in ECV304 was used as stimulator in co-culture with allogeneic T cells, to perform allogeneic T cell proliferation assay and to generate ECV304-specific alloreactive CTL. The proliferation of T cells and the CTL's cytotoxicity against ECV304 were tested by the MTT method. Results The expression of HLA-G1 on the surface of the ECV304 was verified with the immunofluo-rescent staining of the pcDNA3-HLA-G1 transfected cells. The proliferation intensity of allogeneic T cells was significantly decreased after the HLA-G1 expressed in ECV304, as the stimulation index of co-culture of allogeneic T cells with plasmid pcDNA3 transfected ECV304 was 1.59±0. 41, meanwhile it was 1.33±0.46 to pcDNA3-HLA-G1 transfected ECV304, with the difference being significant (P〈0.05). The cytotoxicity of allogeneic CTL was also inhibited by the HLA-G1 expressed on the targets, indicated by the specific lysis of specific CTL against pcDNA3-HLA-G1 transfected ECV304, which was (52.33±4.48)%, meanwhile the specific lysis against plasmid pcDNA3 transfected ECV304 was (64.81±3.07)%, with the difference being significant (P〈0.05). Furthermore, this inhibition was blocked by addition of HLA-G specific mAb (G11E5). Conclusions The expression of HLA-G1 in the ECV-304 cells can inhibit the proliferation and cytotoxicity of allogeneic T cells. Our results support the hypothesis that HLA-G1 bears the tolerance-inducing effects in allogeneic T cell reaction. It is hopeful that HLA-G1 provides an alternative tool to treat autoimmune diseases and
作者 张彩娥 邓云华 吴雄文 韩军艳 杨惠军 梁智辉 龚非力 ZHANG Cai-e, DENG Yun-hua, WU Xiong-wen, et al. (Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China)
出处 《中华器官移植杂志》 CAS CSCD 北大核心 2005年第12期 713-715,共3页 Chinese Journal of Organ Transplantation
基金 中国科学院资助项目
关键词 HLA 内皮 血管 T淋巴细胞 HLA Endothelium, vascular T-lymphocytes
作者简介 通讯作者;Email:xiongwenwu@hotmail.com
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  • 1张彩娥,韩军艳,杨惠军,吴雄文,黄亚非,梁智辉,龚非力.血管内皮细胞(ECV304)表达的HIA-G1对NK细胞杀伤活性的抑制作用[J].细胞与分子免疫学杂志,2005,21(6):676-678. 被引量:3
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