[通过转染猪内皮细胞研究人类白细胞抗原G1减轻异种细胞排斥反应]

[Study on human leukocyte antigen G1 reducing xeno-cell-rejection by transfecting porcine endothelial cells].

作者信息

Xia Zhenxiong, Han Junyan, Wang Shusen, Zeng Ronghua, Zhu Min, Qi Honggang, Chen Dong, Wu Xiongwen, Gong Feili, Chen Shi

机构信息

Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, P. R. China.

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2005 Jun;19(6):486-9.

PMID:16038470

Abstract

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 51Cr-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.

摘要

目的

研究转染HLA - G1的猪内皮细胞（PECs）系是否能改变人外周血单个核细胞（PBMC）和自然杀伤细胞92（NK - 92）介导的细胞溶解作用。

方法

利用脂质体包裹，将携带HLA - G1的pcDNA3.0真核表达载体转染至PECs。采用间接免疫荧光和RT - PCR检测法，检测PECs中HLA - G1的表达情况。通过51Cr释放试验检测PBMC和NK - 92介导的细胞溶解作用的变化。

结果

转染HLA - G1后，可在PECs中检测到HLA - G1在蛋白质和RNA水平的表达。还发现，当转染的PECs和未转染的PECs面对人PBMC和NK - 92介导的细胞溶解作用时，转染的PECs的存活率远高于未转染的PECs。转染后，在转染的PECs中可检测到HLA - G1的表达，且PBMC和NK - 92对PECs介导的细胞溶解作用明显降低（P<0.05）。