HLA-G反义基因增强免疫效应细胞杀伤活性的研究  

Enhancement of Killer Activity of Immune Effector Cells Caused by HLA-G Antisense Oligodeoxynucleotides

作　　者：温见燕[1] 张玲[1] 王芸[1] 毛海婷[1] 温培娥[1] 崔树龄[1] 李登华[1] 顾洪涛[1] 

机构地区：[1]山东省医学科学院基础医学研究所,济南250062

出　　处：《中国肿瘤生物治疗杂志》2004年第2期124-128,共5页Chinese Journal of Cancer Biotherapy

基　　金：山东省自然科学基金资助项目(Y2001C08)

摘　　要：目的:研究HLA—G反义寡核苷酸逆转肿瘤细胞免疫逃逸,提高免疫效应细胞识别杀伤活性的作用和机制。方法:采用反义核酸技术合成HLA-G反义寡核苷酸(ASODN),硫代化修饰与脂质体形成复合物,转导入高表达HLA—G的绒毛膜癌细胞系JEG-3。采用RT-PCR方法检测HLA—G mRNA表达水平的变化;流式细胞术检测细胞表面HLA—G蛋白表达水平的变化;MTT法检测:NK杀伤活性、CD3AK杀伤、增殖活性的变化;ELISA方法探讨ASODN调节CD3AK细胞因子IFN-γ产生的变化。结果:HLA-G ASODN可显著抑制HLA—G mRNA和蛋白水平的表达,逆转可溶型HLA—G分子对NK细胞杀伤活性的抑制作用;可部分逆转HLA—G分子对CD3AK细胞产生IFN-γ的抑制作用,并增强CD3AK的增殖活性,增强JEG-3细胞对CD3AK的杀伤敏感性。结论:HLA—G ASODN通过抑制肿瘤细胞HLA—G mRNA和蛋白水平的表达,增强NK,CD3AK的杀伤作用和机制,发挥逆转肿瘤细胞免疫逃逸作用。Objective: To explore the mechanism of HLA-G antisense oligodeoxynucleotides ( ASODN) on reversion of tumour immune evasion and enhancement of killer activity of immune effector cells. Methods: The expression of HLA-G in JEG-3 cell line was studied by RT-PCR and flow cytometry. MTT assay was used in our studies to detect the change of NK and CD3AK killer activity after ASODN treatment. The effect of IFN-γ secreted by CD3AK after coculture of tumour-lymphocytes was explored by ELISA. Results: HLA-G ASODN could obviously inhibit the expression of HLA-G mRNA and protein. The supernatant of JEG-3 treated by ASODN could reverse the inhibiton of soluble HLA-G molecules to NK killer activity. On the other hand, HLA-G ASODN also could reverse the inhibition of HLA-G to the production of IFN-γ secreted by CD3AK and enhanced the proliferation of CD3AK. Meanwhile, it also improved the susceptibility of JEG-3 to CD3AK. Conclusion: HLA-G ASODN could inhibits the expression of HLA-G mRNA and protein in tumour cells and partly restore the cytotoxicity of NK and CD3AK cells, and thereby partly reverse immune evasion of tumour cells caused by souble HLA-G molecules.

关 键 词：反义寡核苷酸 HLA-G 肿瘤免疫逃逸 JEG-3 

分 类 号：R730.5[医药卫生—肿瘤]