Li Haitao, Zhou Rong, Chen Jingxian, Tian Xingui, Zhang Qiwei, Zeng Qiyi, Gong Sitang
Central Laboratory, Guangzhou Children's Hospital, 318 Renmin Zhong Road, Guangzhou 510120, China.
Vaccine. 2009 Jan 1;27(1):116-22. doi: 10.1016/j.vaccine.2008.10.032. Epub 2008 Oct 31.
Human adenovirus type 3 (HAdV-3) cause respiratory infections globally. There is no safe and efficient HAdV-3 vaccine thus far. In this report, a replication-defective human adenovirus type 3 was constructed by deletion of the whole E1 gene. A HEp-2 cell-based cell line designated HEp-2/E1 was generated and was shown to complement the function of E1 deleted mutant. A partial genome (9.5-100 mu) of HAdV-3 was cloned into pPolyII vector to be a backbone plasmid. A shuttle vector carrying eGFP gene was also constructed. The backbone and shuttle plasmids were co-transfected into the HEp-2/E1 cell line and 293 cell line separately. Typical cytopathic effect (CPE) appeared in the HEp-2/E1, but not in 293 cells 2 weeks after transfection. Wild-type HAdV-3 is neutralized by the sera from the mice immunized with the recombinant HAdV-3. The results demonstrated that the recombinant adenovirus type 3 may serve as a HAdV-3 vaccine candidate.
3型人腺病毒(HAdV-3)在全球范围内引发呼吸道感染。迄今为止,尚无安全有效的HAdV-3疫苗。在本报告中,通过缺失整个E1基因构建了一种复制缺陷型3型人腺病毒。生成了一种基于人喉表皮样癌细胞(HEp-2)的细胞系,命名为HEp-2/E1,其被证明可补充E1缺失突变体的功能。将HAdV-3的部分基因组(9.5-100 mu)克隆到pPolyII载体中作为骨架质粒。还构建了携带绿色荧光蛋白(eGFP)基因的穿梭载体。将骨架质粒和穿梭质粒分别共转染到人喉表皮样癌细胞系HEp-2/E1和293细胞系中。转染2周后,人喉表皮样癌细胞系HEp-2/E1中出现典型的细胞病变效应(CPE),而293细胞中未出现。野生型HAdV-3被用重组HAdV-3免疫的小鼠血清中和。结果表明,重组3型腺病毒可作为HAdV-3疫苗候选物。