人类白细胞抗原G1转染猪内皮细胞降低异种细胞排斥反应的研究  

STUDY ON HUMAN LEUKOCYTE ANTIGEN G1 REDUCING XENO-CELL-REJECTION BY TRANSFECTING PORCINE ENDOTHELIAL CELLS

作　　者：夏振雄[1] 韩军艳[2] 王树森[1] 曹荣华[1] 朱珉[1] 祁洪刚[1] 陈栋[1] 吴雄文[2] 龚非力[2] 陈实[1] 

机构地区：[1]华中科技大学同济医学院器官移植研究所,武汉430030 [2]华中科技大学同济医学院免疫学教研室,武汉430030

出　　处：《中国修复重建外科杂志》2005年第6期486-489,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家高技术研究发展计划(863)资助项目(2001AA216071)~~

摘　　要：目的研究转染人类白细胞抗原G1(humanleukocyteantigenG1,HLA-G1)基因后的猪内皮细胞系(porcineendothelialcells,PECs)细胞,对人外周血单个核细胞(peripheralbloodmononuclearcell,PBMC)和自然杀伤细胞92(naturekillercell92,NK-92)细胞介导的杀伤作用的改变。方法利用脂质体包裹的方法,将携带HLA-G1基因的真核表达载体pcDNA3.0转染PECs细胞,在蛋白质水平通过间接免疫荧光法、在RNA水平通过RT-PCR,检测HLA-G1的表达;取6名健康志愿者,每人5ml静脉血,分离出单个核细胞和NK-92作为效应细胞,利用51Cr释放试验来检测其杀伤作用的改变。结果转染HLA-G1基因后,在转染的内皮细胞中蛋白质水平和RNA水平均检测到HLA-G1基因的表达;PBMC和NK-92细胞介导的对PECs细胞的杀伤作用均明显降低,差异有统计学意义(P<0.05)。结论转染HLA-G1后的PECs,可有效地降低由人NK细胞介导的杀伤作用,从而为抑制异种细胞排斥反应提供了新的思路。Objective To study whether the porcine endothelial cells (PECs) lines transfected by HLA-G1 can alter the lysis mediated by human peripheral blood mononuclear cell (PBMC) and natural killer cell 92(NK-92). Methods By use of liposomes pack, the pcDNA3.0 eukaryotic expression vector carrying HLA-G1 was transfected into PECs. Using indirect immunofluorescence and RT-PCR assays, the HLA-G1 expression in PECs was detected. The alteration of the lysis mediated by PBMC and NK-92 was detected (by ^(51)Cr-release) assays. Results HLA-G1 expression could be detected in PECs after transfection of HLA-G1 at the levels of protein and RNA. It also could be found that the survival rate of transfected PECs was much higher than that of non-transfected PECs, when both of them faced the lysis mediated by human PBMC and NK-92.After transfecting the expression of HLA-G1 could be found in the transfected PECs and the lysis mediated by PBMC and NK-92 to PECs decreased obviously (P<0.05). Conclusion The (PECs) transfected by HLA-G1 can decrease the NK lysis, so that it may provide us a new thought to inhibit the xeno-cell-rejection.

关 键 词：异种排斥反应 猪内皮细胞系 人类白细胞抗原G1 

分 类 号：R392[医药卫生—免疫学]