土拨鼠肝炎病毒和乙型肝炎病毒的核心蛋白存在共同线性B淋巴细胞表位  

Common linear B-cell epitopes in human hepatitis B virus core protein and woodchuck hepatitis virus core protein

作　　者：张振华[1] 王蓉[1] 田拥军[1] 李磊[1] 夏剑波[1] 龚劲松[2] 陆蒙吉[3] 龚非力[4] 杨东亮[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院临床免疫研究室,武汉430030 [2]华中科技大学同济医学院附属同济医院实验医学中心,武汉430030 [3]华中科技大学同济医学院微生物教研室 [4]华中科技大学同济医学院免疫学研究所

出　　处：《中华肝脏病杂志》2007年第8期605-608,共4页Chinese Journal of Hepatology

基　　金：国家自然科学基金（30170889,30271170）;国家“973”计划项目（2001CB510008、2005CB522900）;国家“十五”攻关项目（2001BA705805）

摘　　要：目的寻找并证实土拨鼠肝炎病毒核心抗原（WHcAg）和HBcAg存在共同的B淋巴细胞表位。方法用变性重组WHcAg和HBcAg免疫Balb/c小鼠以制备单克隆抗体；用单克隆抗体对WHcAg和HBcAg进行ELISA、Western blot及免疫组织化学对比分析,同时用该抗体对WHcAg和HBcAg进行表位分析,并在氨基酸水平进行比对。结果经过ELISA、Western blot、免疫组织化学对比分析,制备的单克隆抗体中有2株与HBcAg和WHcAg有交叉反应,而且灵敏度和特异性相似；针对的抗原表位分别位于HBcAg和WHcAg的1～8位氨基酸和125～140位氨基酸。结论WHcAg和HBcAg存在2个共同的线性B淋巴细胞表位,分别位于1～8位和125～140位氨基酸。Objective To search for and verify some common B cell epitopes in the core proteins of woodchuck hepatitis virus and human hepatitis B virus. Methods Monoclonal antibodies against both core proteins of woodchuck hepatitis virus （WHV） and human hepatitis B virus （HBV） were prepared by inoculating Balb/c mice with denatured recombination WHV and HBV core proteins. ELISA and immunoblotting assays for WHcAg and HBcAg were carried out by using these antibodies. Immunohistochemistry was cartied out with liver tissue sections of both WHV-infected woodchucks and chronic HBV-infected patients. The epitopes were mapped with the mouse mAbs （6D1 and 1H4） by using a panel of 24 16mer overlapping peptides coveting the entire WHcAg. The amino acid sequences of WHcAg and HBcAg were compared. Results Cross-reactions were observed between mAbs （6D1 and 1H4） and WHcAg and between Mabs and HBcAg/HBcAg in ELISA and immunoblotting assay. Liver tissue sections of both WHV-infected woodchucks and chronic HBV-infected patients could be stained specifically by mAbs. The epitopes were mapped at aal-8 （6D1） and aa125-140 （1H4） of the core proteins of both WHV and HBV by using ELISA assay. WHcAg and HBcAg share similar amino acids sequences at aal-8 and aa125-140 respectively. Conclusion The core proteins of woodchuck hepatitis virus and human hepatitis B virus share common linear B cell epitopes which span aal-8 and aa125-140 respectively.

关 键 词：肝炎核心抗原 乙型 肝炎病毒 B淋巴细胞 表位 土拨鼠 

分 类 号：R686[医药卫生—骨科学]