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目的 :制备戊型肝炎病毒 (HEV)缅甸株和墨西哥株ORF2重组蛋白 (p16 6Bur和p16 6Mex)的单克隆抗体(McAbs) ,用于分析HEV不同基因型B细胞抗原表位的特点。方法 :将免疫BALB C小鼠脾细胞与SP2 0骨髓瘤细胞融合 ,获得分泌抗 p16 6Bur和抗 p16 6MexMcAbs的杂交瘤细胞株 ,然后采用ELISA和免疫印迹法测定McAbs与不同基因型HEVORF2编码蛋白p16 6的免疫反应性。结果 :获得 4株杂交瘤细胞株 ,即分泌抗 p16 6BurMcAbs的 2C2 、2B1 以及分泌抗 p16 6MexMcAbs的D8G1 0 和E5E1 2 ,其中 2B1 分泌的McAb仅能与第Ⅰ、Ⅱ基因型编码的重组蛋白结合 ,而其余 3株分泌的McAbs既能与I、II基因型HEV的p16 6重组蛋白发生反应 ,也能与III、IV基因型的p16 6蛋白反应。结论 :HEV第Ⅰ、Ⅱ基因型与Ⅲ、Ⅳ基因型ORF2编码蛋白既有共同又有不同的B细胞抗原表位。
OBJECTIVE: To prepare McAbs of hepatitis E virus (HEV) strain Myanmar and ORF2 recombinant proteins of Mexico strain (p16 6Bur and p16 6Mex) for analyzing the characteristics of B cell epitopes of different genotypes of HEV. Methods: The spleen cells of immunized BALB C mice were fused with SP2 0 myeloma cells, and the hybridoma cell lines secreting anti-p16 6Bur and anti-p16 6MexMcAbs were obtained. ELISA and Western blotting were used to determine the relationship between McAbs and HEVORF2-encoding proteins p16 6 immunoreactivity. RESULTS: Four hybridoma cell lines, 2C2 and 2B1 secreting anti-p16 6BurMcAbs and D8G10 and E5E12 secreting anti-p166MexMcAbs were obtained. The McAbs secreted by 2B1 could only bind to the recombinant proteins encoded by genotypes I and II , Whereas McAbs secreted by the remaining 3 strains both reacted with the p166 recombinant protein of the HEV of the I and II genotypes and also reacted with the p166 proteins of the III and IV genotypes. Conclusion: The genotypes Ⅰ and Ⅱ of HEV have both common and different B cell epitopes with ORF2 encoding proteins of genotype Ⅲ and Ⅳ.