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目的:证实HLA-G1分子能够抑制NK细胞对同种血管内皮细胞系的杀伤作用。方法:采用脂质体介导的DNA转染技术,以构建的真核表达质粒pcDNA3-HLA-G1转染人脐静脉内皮细胞系ECV304,再用免疫荧光法检测表达的HLA-G1分子。并用MTT比色法检测HLA-G1对NK细胞杀伤活性的影响。结果:ECV304细胞上可稳定表达HLA-G1。NK细胞对空质粒pcDNA3转染的ECV304细胞的杀伤率为(50.6±18.1)%;而对pcDNA3-HLA-G1转染的ECV304细胞的杀伤率为(29.7±11.4)%,两者差异具有显著意义(P<0.01)。结论:HLA-G1分子可明显抑制NK细胞对同种血管内皮细胞的杀伤效应。
Objective: To confirm that HLA-G1 molecules can inhibit NK cell killing of allogeneic vascular endothelial cell line. Methods: The human umbilical vein endothelial cell line ECV304 was transfected with the eukaryotic expression plasmid pcDNA3-HLA-G1 by using liposome-mediated DNA transfection technique. The expressed HLA-G1 molecules were detected by immunofluorescence. MTT assay was used to detect the effect of HLA-G1 on NK cell killing activity. Results: HLA-G1 was stably expressed on ECV304 cells. The killing rate of NK cells to ECV304 cells transfected with empty plasmid pcDNA3 was (50.6 ± 18.1)%, while the killing rate of ECV304 cells transfected with pcDNA3-HLA-G1 cells was (29.7 ± 11.4)%, the difference was significant Significance (P <0.01). Conclusion: HLA-G1 can obviously inhibit the killing effect of NK cells on allogeneic vascular endothelial cells.