摘要
目的 :探讨HBV野生株 (WT)及核壳蛋白变异株L97、V6 0上调HepG2细胞表面HLA I表达的机制。方法 :将已构建的重组表达载体EBO WT、EBO L97及EBO V6 0 ,分别经脂质体介导转染HepG2细胞 ,以空载体EBO作为对照。用RT PCR半定量法 ,检测细胞内HLA A基因及抗原提呈相关基因LMP2、TAP1和tapasinmRNA的表达 ;用Westernblot测定细胞内HLA I蛋白的表达。结果 :3株HBV重组表达载体转染的细胞 ,均呈现明显的HLA AcDNA的PCR扩增条带及HLA I蛋白条带 ,但其条带的强度有差异 ,EBO L97、EBO WT和EBO V6 0依次减弱。TAP1cDNA扩增带清晰 ,3株HBV间无明显差别。对照细胞未呈现HLA A的条带和仅呈现微弱的TAP1扩增带。各转染细胞均未检出LMP2及tapasinmRNA的表达。 结论 :HBV能诱导HepG2细胞内HLA I分子的合成量 ,使TAP1基因转录增强 ,HLA I的表达上调。核壳蛋白变异株L97和V6 0 ,可使宿主细胞内HLA
AIM: To explore the mechanism of up-regulation of HLA-I expression on HepG2 cells by wild type (WT) and nucleocapsid mutants (L97 and V60) of hepatitis B virus (HBV). METHODS: The HBV-stable expression vectors EBO-WT, EBO-L97 and EBO-V60 were transfected into HepG2 cells via the liposome mediation, respectively. The cells were assayed by semi-quantitative RT-PCR for HLA-A gene and antigen presentation-related genes LMP2, TAP1, and tapasin mRNA expression. Western blot was applied for analysis of HLA-I protein expression in the cells. RESULTS: HepG2 cells transfected by the 3 HBV expression vectors expressed HLA-A and TAP1, while there was no expression of HLA-A and only marginal expression of TAP1 in HepG2 cells transfected by control vector EBO. The expression level of HLA-A in the transfected cells decreased successively in the order of EBO-L97, EBO-WT and EBO-V60. There was no significant difference in the expression level of TAP1 between HepG2 cells transfected by the 3 HBV expression vectors. No detectable expression of LMP2 and tapasin was observed for all transfected cells. CONCLUSION: HBV can induce the expression of HLA-I molecule and TAP1 in HepG2 cells.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第1期74-77,共4页
Chinese Journal of Cellular and Molecular Immunology
