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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 the expression of porcine endothelial cell (PECs) in human peripheral blood mononuclear cells (PBMCs) and natural killer cell line 92 (NK cells) after transfecting human leukocyte antigen G 1 (HLA-G1) -92) Changes in cell-mediated killing. Methods The eukaryotic expression vector pcDNA3.0 carrying HLA-G1 gene was transfected into PECs cells by liposome-encapsulation method. The level of HLA-G1 was detected by RT-PCR at the protein level by indirect immunofluorescence Expression; Take 6 healthy volunteers, 5ml venous blood per person, isolated mononuclear cells and NK-92 as effector cells, 51Cr release test to detect changes in its killing effect. Results After HLA-G1 gene transfection, the expression of HLA-G1 gene was detected in the transfected endothelial cells at both protein and RNA levels. The cytotoxicity of PBMC and NK-92 cells to PECs was significantly decreased, The difference was statistically significant (P <0.05). Conclusion The PECs transfected with HLA-G1 can effectively reduce the cytotoxicity mediated by human NK cells, which provides a new idea for inhibiting the rejection of xenogeneic cells.