【摘要】 利用非复制痘苗病毒质粒载体 pNEOCK11β75及 pNEOCK ,改造了表达EB病毒主要膜蛋白 gp35 0 /2 2 0的复制型重组痘苗病毒VMA ,构建了非复制型重组痘苗病毒VMAΔCK。该病毒能在鸡胚原代成纤维细胞 (CEF)中正常繁殖 ,而在人源细胞中不能正常繁殖。在CEF中连续传代至第 2 5代 ,经PCR证明 ,该病毒符合非复制型重组痘苗病毒的特征。经免疫荧光及免疫酶斑法证实 ,VMAΔCK可稳定表达gp35 0 /2 2 0 ,且表达水平与VMA无明显差异。VMAΔCK经腹腔免疫Balb/C小鼠 ,4周后能诱生一定水平的抗 gp35 0 /2 2 0特异性抗体 ,加强免疫 2周后该抗体水平明显升高。这一结果类似于VMA免疫Balb/C小鼠的结果。初免后 ,VMAΔCK免疫组抗痘苗抗体水平明显低于VMA免疫组 ;加强免疫 2周后 ,两组小鼠的抗痘苗抗体水平趋于一致。上述结果证明 ,所构建的非复制痘苗病毒不影响目的抗原的表达 ,也不影响该抗原的免疫原性 ,但导致病毒毒力下降 ,而且用该病毒免疫小鼠后小鼠抗痘苗病毒载体的免疫反应明显下降。更多还原

【Abstract】 The non-replicating recombinant vaccinia virus VMAΔCK expressing Epstein-Barr virus major membrane antigen gp350/220 was constructed using non-replicating vaccinia virus plasmid vector pNEOCK11β75 and pNEOCK. The non-replicating virus could reproduce normally in CEF cells, but not in cells originated from man. Further investigation by IFA, Dot-ELISA demonstrated that the non-replicating virus could steadily express gp350/220. There were no differences in expressing gp350/220 in the replicating and non-replicating viruses. By abdominal cavity immunization, the special IgG antibodies against gp350/220 could be induced by VMAΔCK in BalB/c mice after 4 weeks, the IgG titer could be boosted about 4 times after revaccination two weeks later, which reached a similar level to that induced by the replicating virus VMA. These results confirmed that the non-replicating vaccinia virus did not affect the expression and immunogenicity of the expected antigen, but decreased the virulence of the vaccinia virus and the immunoreactivity against the vaccinia virus vector.更多还原