乙型肝炎病毒多聚酶末端蛋白重链可变区抗体体外抑制病毒的复制  

Variable fragments of heavy chain antibody against the terminal protein region of hepatitis B virus polymerase inhibits the replication of hepatitis B virus in cultured cells

作　　者：于俊岩[1] 汤勃[1] 周吉军[1] 邓春青[1] 兰林[1] 王宇明[1] 

机构地区：[1]第三军医大学西南医院感染科,重庆400038

出　　处：《中华传染病杂志》2007年第1期15-19,共5页Chinese Journal of Infectious Diseases

基　　金：国家自然科学基金资助项目（30100159）

摘　　要：目的用蛋白片段互补法（PCA）选择HBV多聚酶末端蛋白（TP）重链可变区（VH）抗体，研究特异性VH抗体体外抑制HBV的复制与分泌。方法以HBV多聚酶TP区为抗原，用PCA法进行特异性VH抗体筛选。特异性VH抗体基因定向亚克隆入真核表达载体pZeoSV2（＋），构建pZeoSV2（＋）-VH重组质粒，用脂质体介导的转染技术，将其导入HepG2．2．15细胞，观察特异性抗体的生物学活性。结果用PCA法从抗体库中选择出3个特异性抗体：VH1、VH2和VH3，其中与抗原亲和力最高的是VH1。转染pZeoSV2（＋）-VH1的HepG2．2．15细胞比未转染pZeoSV2（＋）-VH1或只转染空载体pZeoSV2（＋）的HepG2．2．15细胞病毒颗粒分泌明显减少，上清液内为2．9787±0．2761比5．1504±0．2761和5．2951±0．2410（P〈0．05）；细胞内为5．2839±0．1629比8．3004±0．3232和8．5321±0．1383（P〈0．05）。结论用PCA法可从抗体库中选择出HBV多聚酶TP区特异性VH抗体，该抗体在体外可抑制HBV的复制与分泌。Objective To study a functional variable fragments of heavy chain（VH） antibody against the terminal protein（TP） region of hepatitis B virus（HBV） polymerase and its inhibition on the replication of HBV in vitro. Methods The TP region of HBV secreted by the HepG 2.2.15 cells was used as an antigen, and the specific antibodies were selected with protein-fragment complementa tion assay （PCA）. The highest-affinity antibody VH 1 gene was sub cloned into pZeoSV2（＋） expression vector, and then eukaryotic expression plasmid pZeoSV2 （＋ ）- VH1 was transfected into the HepG 2.2.15 cells with liposome. Results There were three TP region antigen-specific VH could be directly in vivo selected with PCA. And the HepG 2.2.15 cells were inhibited the replication of HBV by the anti-TP VH antibodies. The contents of HBV DNA in the pZeoSV2（ ＋）-VH1 transfected cells were significantly lower than those in the non-transfected and pZeoSV2（＋） transfected cells. Supernatant： 2. 978 7±0. 276 1 vs 5. 150 4±0. 276 1, 5. 295±0. 241 0 （P〈 0.05） ; intracellular： 5. 283 9 ±0. 1629 vs 8. 300 4±0. 323 2, 8.532 1±0.1383（P〈0.05）. Conclusions The TP region antigen specific VH can be directly in vivo selected with PCA. And the HepG 2.2.15 cells are inhibited the replication of HBV by the anti-TP VH antibodies.

关 键 词：肝炎病毒 乙型 蛋白质片段互补 重链可变区抗体 

分 类 号：R686[医药卫生—骨科学]