乙型肝炎病毒核心启动子缺失变异对病毒抗原表达的影响

Effects of partial deletion in the core promoter of hepatitis B virus genome on viral antigen ex-pressions

目的为研究乙型肝炎病毒(HBV)核心启动子(CP)20/21bp部分缺失(nt1748/1747至nt1767)及同时存在的A1896点变异对病毒抗原表达的影响。方法利用前期构建的HBV全基因的重组载体转染HepG2细胞后,对病毒抗原进行ELISA检测及Western-blotting分析。结果变异株分泌到细胞外的HBsAg、HBeAg及细胞内的HBcAg表达量较野毒株均明显减少。结论HBVCP20/21bp部分缺失株及同时存在的A1896点变异株的病毒抗原表达较野毒株显著下降。

Objective To study the effects of 20/21 bp partial deletion mutation (from nt 1 748 or nt 1 747 to nt 1 767) in the core promoter (CP) region of hepatitis B virus (HBV) genome complicated by precore stop condon mutation at nt 1 896 on the expression of the viral antigens. Methods Eukaryotic expression vector containing full-length HBV genome with the above mutations was constructed. After transfection of the recombinant HBV plasmids into HepG2 cells, the expression of the viral antigens was examined with enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis. Results As shown by ELISA and Western blotting analysis, the amount of extracellular secretion of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) along with intracellular hepatitis B core antigen (HBcAg) in the cells transfected with vectors containing HBV genomes with partial deletion in the CP region was markedly reduced compared with that produced by wild-type HBV. Conclusion The mutations in question causes marked reduction in viral antigen production by HBV in comparison that by wild-type HBV.