摘要
目的 探讨重组腺病毒免疫实验动物后的抗体反应。方法 利用RT-PCR方法,从HAV-L8株的RNA中扩增出结构蛋白vp3+vpl基因,克隆到穿梭质粒pXCX2 NotI上。采用磷酸钙-DNA共沉淀技术,将E1缺失的复制缺陷型腺病毒载体pJM17与pXCX2-CMV-HAV共转染293细胞。结果 通过胞内同源重组,经RT-PCR、免疫荧光染色和蛋白印迹鉴定表明,获得了复制缺陷型重组腺病毒rAdHAV。纯化后的rAdHAV滴度为1×109.0TCID50/mL。昆明种小白鼠口服rAdHAV后,诱导产生了抗HAV IgG。结论 复制缺陷型腺病毒可望成为发展口服病毒疫苗的有效载体系统。
ve To discuss the immune reaction in the animals inoculated with the reeombinant adenovirus. Methods The structural gene (vp3 + vpl) of HAV-L8 was amplified from its RNA by RT-PCR and was cloned into Ad shuttle plasmid (pXCX2 Not I ). A reeombinant replication-defective adenovirus was rescued in 293 packaging cells via homologous recombination in vivo of both plasmid pXCX2-CMV-HAV and pJM17 containing Ad5 genome with deletions of El region. Results A series of methods such as RT-PCR, immunofluorescence and western blot were employed to identify the generated reeombinant adenovirus (rAdHAV). The titer of stocks of rAdHAV was up to 1× 109.0 TCID50/mL. After rAdHAV was delivered to Kunmimg mice by oral administration, anti-HAV IgC was tested in the mice. Conclusions A reeombinant replication-defective adenovirus will become an efficient vector system to develop an oral vims vaccine.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2002年第3期192-200,共9页
Immunological Journal
