可溶性HLA-G1抑制人自然杀伤细胞NK92对猪内皮细胞PED的黏附

Soluble HLA-G1 inhibits xenotransplantation rejection through degrading NK92 to adhere to PED

目的研究可溶性HLA-G1(HLA-G5)对人自然杀伤细胞(NK细胞)黏附到猪血管内皮细胞(PED)的抑制作用,从而降低人NK细胞对猪血管内皮细胞杀伤杀伤活性。方法利用核转染技术将pcDNA3-HLA-G5转入LCL721.221细胞株。以RT-PCR、Dot-ELISA技术分别在基因水平和蛋白水平检测HLA-G5的表达;以人类NK细胞系(NK92)效应细胞,猪内皮细胞系(PED)为靶细胞,检测NK92在静止和流动状态下对PED的黏附作用;四甲基偶氮唑盐(MTT)法检测HLA-G5抑制NK细胞的杀伤活性。结果与未加入HLA-G5的对照组相比,NK92在静止和流动状态下对PED的黏附功能明显降低(P<0.01),且对HLA-G5组PED的杀伤效率均有明显下降(P<0.01)。结论HLA-G5可以通过抑制NK92对PED的黏附功能,来减轻NK92对PED的杀伤作用。

Objective To study the inhibitory effect of soluble HLA-G1 （ HLA-G5 ） on xenotransplantation rejection through degrading the role of NK92 to adhere to PED. Methods The recombinant expression vector pcDNA3-HLA-G5 was transfected into the lymphoblastoid cell line LCL721. 221 by nucleofection. The expression of HLA-G5 in the transfected LCL721. 221 was detected by RT-PCR and Dot-ELISA technique. NK cell line （NK92） was used as NK effect ceils and the porcine aortic endothelial ceils line （PED） as targets. The cytotoxicity of NK92 was analyzed by MTT. Results HLA-G5 conferred significance in degrading the role of NK92 to adhere to PED （P 〈 0.01 ）, and protected PED against NK2 medicated lysis. The rate of NK92 cell eytotoxieity was reduced to 25.5% as compared to 71.2% in the control group （ P 〈 0.01 ）. Conclusion HLA-G5 can degrade the role of NK92 to adhere to PED, and directly protect PED against NK92, indicating that HLA-G5 may be useful to prevent human NK cells responding to porcine xenografts.