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一种重组的复制缺陷型人3型腺病毒:一种候选疫苗。

A recombinant replication-defective human adenovirus type 3: a vaccine candidate.

作者信息

Li Haitao, Zhou Rong, Chen Jingxian, Tian Xingui, Zhang Qiwei, Zeng Qiyi, Gong Sitang

机构信息

Central Laboratory, Guangzhou Children's Hospital, 318 Renmin Zhong Road, Guangzhou 510120, China.

出版信息

Vaccine. 2009 Jan 1;27(1):116-22. doi: 10.1016/j.vaccine.2008.10.032. Epub 2008 Oct 31.

Abstract

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.

摘要

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。

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