戊型肝炎病毒Ⅰ、Ⅱ与Ⅲ、Ⅳ基因型B细胞抗原表位的异同  

Comparison of B cell antigenic epitopes between hepatitis E virus genotypeⅠ,Ⅱ and genotype Ⅲ,Ⅳ by using monoclonal antibodies

作　　者：赵宇[1] 梁久红[1] 孟继鸿[1] 张兰芳[1] 张红梅[1] 丁福[1] 季晓辉[2] 

机构地区：[1]东南大学医学院病原生物学和免疫学系,南京210009 [2]南京医科大学病原生物学和免疫学系,南京210009

出　　处：《中国免疫学杂志》2004年第9期591-595,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金 ( 3 0 2 712 3 1);江苏省卫生厅医学科技发展基金(H2 0 0 115 )资助项目

摘　　要：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 characterize B cell antigenic epitopes of different genotypes of hepatitis E virus (HEV) with monoclonal antibodies (McAbs) against HEV ORF2-encoded proteins (p166) of the HEV Burma and Mexico strains.Methods:The recombinant proteins (p166Bur and p166Mex) were used to immunize BALB/C mice,respectively. Spleen cells of the immunized mice were fused with SP2/0 myeloma cells. Hybridoma clones which secreted McAbs against p166Bur or p166Mex were screened by an enzyme-linked immunosorbent assay (ELISA). Finally, different p166 recombinant proteins (p166Bur, p166Pak, p166Mor, p166Mex, p166Us, p166Nz and p166Ch) obtained from HEV genotype Ⅰ,Ⅱ,Ⅲ or Ⅳ were tested with the generated McAbs by ELISA and Western blotting.Results:Four hybridoma cell lines, numbered 2C 2, 2B 1 and D 8G 10, E 5E 12 were obtained from the spleen cells of mice immunized with p166Bur and p166Mex, respectively. McAb produced by 2B 1 reacted with p166Bur, p166Pak, p166Mor and p166Mex, which derived from HEV genotypeⅠand Ⅱ, but did not react with p166Us, p166Nz and p166Ch, which derived from HEV genotype Ⅲ and Ⅳ. However, McAbs produced by 2C 2 , D 8G 10 and E 5E 12 were reactive to the p166 proteins prepared not only from genotype Ⅰ and Ⅱ, but also from genotype Ⅲ and Ⅳ.Conclusion:Both common and different antigenic epitopes within p166 recombinant proteins exist between HEV genotype Ⅰ, Ⅱ and genotype Ⅲ,Ⅳ.

关 键 词：戊型肝炎病毒 抗原表位 单克隆抗体 重组蛋白 基因型 

分 类 号：R575.1[医药卫生—消化系统] R392.12[医药卫生—内科学]