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【目的】研究人类免疫缺陷病毒(HIV)-1 nef调节细胞表面的人类白细胞抗原(HLA)-I类分子及诱导细胞凋亡情况。【方法】将293T细胞分为两组,用脂质体法将含HIV-1 nef基因的质粒及空载体分别转染细胞,Western blot检测HIV-1 nef在转染细胞中的表达,荧光抗体染色结合流式细胞术检测细胞表面的HLA-I类分子表达及细胞凋亡情况。【结果】HIV-1 nef在转染细胞中有表达。转染HIV-1 nef的细胞与转染空载体的细胞经抗HLA-A,B,C-FITC染色后,平均荧光强度分别为205±22及269±15,两者相比有显著性差异(P<0.01);经Annexin V-APC/PI染色后,阳性细胞百分率分别为(60.82±8.32)%及(8.12±5.43)%,两者相比亦有显著性差异(P<0.01)。【结论】HIV-1 nef下调细胞表面的HLA-I类分子及诱导细胞凋亡。
【Objective】 To investigate the effect of human immunodeficiency virus (HIV) -1 nef on the regulation of human leukocyte antigen (HLA) -I on cell surface and the induction of apoptosis. 【Methods】 293T cells were divided into two groups. The plasmids and empty vector containing HIV-1 nef gene were transfected into the cells by lipofectamine respectively. The expression of HIV-1 nef in transfected cells was detected by Western blot. Staining and flow cytometry were used to detect the expression of HLA class I molecules and cell apoptosis on the cell surface. 【Results】 HIV-1 nef was expressed in transfected cells. The average fluorescence intensity of cells transfected with HIV-1 nef and those transfected with empty vector was 205 ± 22 and 269 ± 15 after anti-HLA-A, B and C-FITC staining, respectively (P <0.01). After Annexin V-APC / PI staining, the percentages of positive cells were (60.82 ± 8.32)% and (8.12 ± 5.43)%, respectively. There was also significant difference between the two groups (P <0.01). 【Conclusion】 HIV-1 nef downregulates HLA-I molecules on cell surface and induces apoptosis.