Genetic stability of a recombinant adenovirus vaccine vector seed library expressing human papillomavirus type 16 E6 and E7 proteins
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- Published online on: February 5, 2015 https://doi.org/10.3892/etm.2015.2268
- Pages: 1161-1165
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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 6.31x109 IU/ml and 3.0x109 IU/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 109 IU/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.
