title: Genetic stability of a recombinant adenovirus vaccine vector seed library expressing human papillomavirus type 16 E6 and E7 proteins.
abstract: The aim of the present study was to understand the genetic stability of a master seed bank (MSB) and a working seed bank (WSB) of an adenovirus vector vaccine expressing the human papillomavirus (HPV) type 16 E6 and E7 fusion proteins (Ad-HPV16E6E7). Microscopic examination and viral infectious efficacy were used to measure the infectious titers of the Ad-HPV16E6E7 MSB and WSB. Polymerase chain reaction was used to analyze the stability of the Ad-HPV16E6E7 target gene insertion, while western blot analysis and immunofluorescence were used to assess the expression levels of the Ad-HPV16E6E7 target protein. A C57BL/6 mouse TC-1 tumor cell growth inhibition model was used to evaluate the biological effect of Ad-HPV16E6E7 administration. The infectious titers of the Ad-HPV16E6E7 MSB and WSB were 6.31×109IU/ml and 3.0×109IU/ml, respectively. In addition, the expression levels of the inserted target genes and target proteins were found to be stable. In the mouse TC-1 tumor inhibition analysis, when the virus titers of the Ad-HPV16E6E7 MSB and WSB were 109IU/ml, the tumor inhibition rate was 100%, which was significantly different when compared with the control group (χ2MSB=20.00 and χ2WSB=20.00; P<0.01). Therefore, the Ad-HPV16E6E7 vaccine seed bank is genetically stable and meets the requirements for vaccine development.
authors.affiliation: Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310053, P.R. China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wu
authors.affiliation: Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310053, P.R. China.
authors.forename: Ke-DA
authors.initials: KD
authors.lastname: Chen
authors.affiliation: Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310053, P.R. China.
authors.forename: Meng
authors.initials: M
authors.lastname: Gao
authors.affiliation: Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310053, P.R. China.
authors.forename: Gang
authors.initials: G
authors.lastname: Chen
authors.affiliation: Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310053, P.R. China.
authors.forename: Su-Feng
authors.initials: SF
authors.lastname: Jin
authors.affiliation: Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310053, P.R. China.
authors.forename: Fang-Cheng
authors.initials: FC
authors.lastname: Zhuang
authors.affiliation: Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310053, P.R. China.
authors.forename: Xiao-Hong
authors.initials: XH
authors.lastname: Wu
authors.affiliation: Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310053, P.R. China.
authors.forename: Yun-Shui
authors.initials: YS
authors.lastname: Jiang
authors.affiliation: Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310053, P.R. China.
authors.forename: Jian-Bo
authors.initials: JB
authors.lastname: Li
keywords: adenovirus vector; genetic stability; human papillomavirus type 16; seed bank