Construction of Prophylactic Human Papillomavirus Type 16 L1 Capsid Protein Vaccine Delivered by Live Attenuated Shigella flexneri Strain sh42  

作　　者：Xiao-Feng YANG Xin-Zhong QU Kai WANG Jin ZHENG Lu-Sheng SI Xiao-Ping DONG Yi-Li WANG 

机构地区：[1]Key Laboratory of Biomedical Information Engineering of Ministry of Education, Institute for Cancer Research, Xi＇an Jiaotong University, Xi＇ an 710061, China [2]National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100032, China [3]Department of Obstetrics and Gynecology, First Hospital of Xi＇an Jiaotong University, Xi＇an 710061, China

出　　处：《Acta Biochimica et Biophysica Sinica》2005年第11期743-750, ,共8页生物化学与生物物理学报（英文版）

基　　金：This work was supported by the grants from the National High Technology Research and Development Program of China(No.2001AA215221);the National Natural Science Foundation of China(No.30271184)

摘　　要：To express human papillomavirus （HPV） L 1 capsid protein in the recombinant strain of Shigella and study the potential of a live attenuated Shigella-based HPV prophylactic vaccine in preventing HPV infection, the icsA/virG fragment of Shigella-based prokaryotic expression plasmid pHS3199 was constructed. HPV type 16 L1 （HPV16L1） gene was inserted into plasmid pHS3199 to form the pHS3199-HPV16L1 construct, and pHS3199-HPV 16L1 was electroporated into a live attenuated Shigella strain sh42. Western blotting analysis showed that HPV 16L 1 could be expressed stably in the recombinant strain sh42-HPV 16L 1. Sereny test results were negative, which showed that the sh42-HPV16L1 lost virulence. However, the attenuated recombinant strain partially maintained the invasive property as indicated by the HeLa cell infection assay. Specific IgG, IgA antibody against HPV16L1 virus-like particles （VLPs） were detected in the sera, intestinal lavage and vaginal lavage from animals immunized by sh42-HPV 16L1. The number of antibodysecreting cells in the spleen and draining lymph nodes were increased significantly compared with the control group. Sera from immunized animals inhibited mufine hemagglutination induced by HPV 16L1 VLPs, which indicated that the candidate vaccine could stimulate an efficient immune response in guinea pig＇s mucosal sites. This may be an effective strategy for the development of an HPV prophylactic oral vaccine.To express human papillomavirus （HPV） L 1 capsid protein in the recombinant strain of Shigella and study the potential of a live attenuated Shigella-based HPV prophylactic vaccine in preventing HPV infection, the icsA/virG fragment of Shigella-based prokaryotic expression plasmid pHS3199 was constructed. HPV type 16 L1 （HPV16L1） gene was inserted into plasmid pHS3199 to form the pHS3199-HPV16L1 construct, and pHS3199-HPV 16L1 was electroporated into a live attenuated Shigella strain sh42. Western blotting analysis showed that HPV 16L 1 could be expressed stably in the recombinant strain sh42-HPV 16L 1. Sereny test results were negative, which showed that the sh42-HPV16L1 lost virulence. However, the attenuated recombinant strain partially maintained the invasive property as indicated by the HeLa cell infection assay. Specific IgG, IgA antibody against HPV16L1 virus-like particles （VLPs） were detected in the sera, intestinal lavage and vaginal lavage from animals immunized by sh42-HPV 16L1. The number of antibodysecreting cells in the spleen and draining lymph nodes were increased significantly compared with the control group. Sera from immunized animals inhibited mufine hemagglutination induced by HPV 16L1 VLPs, which indicated that the candidate vaccine could stimulate an efficient immune response in guinea pig＇s mucosal sites. This may be an effective strategy for the development of an HPV prophylactic oral vaccine.

关 键 词：human papillomavirus type 16 attenuated Shigella flexneri VACCINE cervical cancer PROPHYLACTIC 

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