Wang S S, Han J Y, Wu X W, Cao R H, Qi H G, Xia Z X, Chen D, Gong F L, Chen S
Institute of Organ Transplantation, Key Laboratory appointed by China Ministry of Education and China Ministry of Health, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Transplant Proc. 2004 Oct;36(8):2473-4. doi: 10.1016/j.transproceed.2004.08.048.
Human natural killer (NK) cells, which can directly lyse porcine endothelial cells, play an important role in xenotransplantation. HLA-G is a nonclassical major histocompatibility complex (MHC) class I molecules that has been implicated in protecting susceptible target cells from lysis by NK cells. The objective was to study the effect of protecting porcine endothelial cells transfected with HLA-G1 from human NK cell lysis.
The recombinant expression vector pcDNA3-HLA-G1 was transfected into primary cultured porcine aortic endothelial cells (PAECs) by lipofection. Surface expression of HLA-G1 in transected PAECs was confirmed by an immunofluoresence technique. Peripheral blood mononuclear cells (PBMC) and NK cell line (NK92) were used as NK effects cells with pcDNA3-HLA-G1-transfected PAECs as targets in a MTT method using pcDNA3 transfection as a negative control.
Expression of HLA-G1 on PAECs conferred significant protection against NK-mediated lysis. The rate of NK92 cytotoxicity was reduced to 41.5% +/- 14.0% from 75.3% +/- 10.5% in the control group (P < .01). Similarly the rate of the PBMC cytotoxicity among different donors (n = 7) was reduced to 45.4% +/- 12.1% in contrast to 74.6% +/- 11.2% in the control group (P < .05).
HLA-G1 molecules can directly protect xenogeneic PAECs against attack by human NK cells. These results indicate that the expression of HLA-G1 on the porcine cell surface may provide a new approach to overcome NK-mediated immunity to xenografts.
人类自然杀伤（NK）细胞可直接裂解猪内皮细胞，在异种移植中起重要作用。HLA - G是一种非经典的主要组织相容性复合体（MHC）I类分子，与保护易感靶细胞免受NK细胞裂解有关。目的是研究保护转染HLA - G1的猪内皮细胞免受人类NK细胞裂解的效果。
通过脂质体转染将重组表达载体pcDNA3 - HLA - G1转染至原代培养的猪主动脉内皮细胞（PAECs）。通过免疫荧光技术确认转染的PAECs中HLA - G1的表面表达。以pcDNA3转染作为阴性对照，采用MTT法，将外周血单个核细胞（PBMC）和NK细胞系（NK92）用作NK效应细胞，以pcDNA3 - HLA - G1转染的PAECs作为靶细胞。
PAECs上HLA - G1的表达赋予了对NK介导的裂解的显著保护作用。NK92细胞毒性率从对照组的75.3%±10.5%降至41.5%±14.0%（P <.01）。同样，不同供体（n = 7）的PBMC细胞毒性率从对照组的74.6%±11.2%降至45.4%±12.1%（P <.05）。
HLA - G1分子可直接保护异种PAECs免受人类NK细胞的攻击。这些结果表明，猪细胞表面HLA - G1的表达可能为克服NK介导的异种移植免疫提供一种新方法。
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