HLA-G慢病毒转染抑制NK细胞毒功能的实验研究

Expression of HLA-G molecule on HEC-I-B cell and inhibit NKcytotoxicity activity

目的建立-株稳定表达人类白细胞抗原G（human leucocyte antigenG，HLA—G）的子宫内膜癌细胞系，探讨HLA—G对NK92细胞的杀伤活性影响。方法将HLA—G慢病毒包装并转染于HEC-1-B细胞系，稳定转染HLA—G基因的I-IEC-1-B与NK92细胞系共培养，并使用乳酸脱氢酶（1actate dehydrognase，LDH）释放法检测HLA—G表达对自然杀伤（naturalkiller，NK）92细胞杀伤活性的影响。结果外源基因HLA—G成功介导转染在人子宫内膜癌HEC-1-B细胞系，通过免疫荧光可见HLA—G表达率为95％，乳酸脱氢酶（LDH）释放法检测HLA—G基因表达能够有效抑制NK92细胞的杀伤活性，与对照组相比有统计学意义（P〈0．05）。结论HLA—G是-个重要的免疫抑制分子，稳定表达HLA—G的子宫内膜癌细胞可明显抑制NK92细胞的杀伤活性，可能诱导移植免疫耐受的发生。

Objective To establish a stable expression of human leucocyte antigen G （HLA-G） on the human endometrial carcinoma （HEC-1-B） cell line by lentivirus transfection and study on its inhibition of natural killer （NK）-92 cytoto xicity activity. Methods HLA-G was packaged by lentivirus transfection on the HEC-1-B ceils. The HEC-1-B ceils with stable expression I-ILA-G were co-cultured with NK92 cells, and NK cytotoxicity was detected by lactate dehydrognase （LDH）method. Results HLA-G was successfully transfected into the HEC-1-B cells, and the transfection efficiency was 95% by immunofluorescence. As compared with the control, HLA-G expression on HEC-1-B can effectively inhibit the cytotoxicity of NK92 cells （P〈 0.05）. Conclusion The expression of HLA-G on the HEC-1-B cells can significantly inhibit the cytotoxicity of NK cells. It may play a role in transplantation immune tolerance.