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Construction of Prophylactic human papillomavirus type 16 L1 Capsid Protein Vaccine Delivered by Live Attenuated Shigella flexneri Strain sh42

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Acta Biochimica et Biophysica Sinica 2005年第11期37卷 743-750页

作者： Xiao-Feng YANG Xin-Zhong QU Kai WANG Jin ZHENG L(?)-Sheng SI Xiao-Ping DONG Yi-Li WANG Key Laboratory of Biomedical Information Engineering of Ministry of Education,Institute for Cancer Research,Xi'an Jiaotong University,Xi'an 710061.China National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 100052,China Department of Obstetrics and Gynecology,First Hospital of Xi'an Jiaotong University,Xi'an 710061,China Key Laboratory of Biomedical Information Engineering of Ministry of Education Institute for Cancer Research Xi'an Jiaotong UniversityXi'an 710061 China Department of Obstetrics and Gynecology First Hospital of Xi'an Jiaotong UniversityXi'an 710061 China National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and PreventionBeijing 100052 China

To express human papillomavirus(HPV)L1 capsid protein in the recombinant strain of Shigellaand study the potential of a live attenuated Shigella-based HPV prophylactic vaccine in preventing HPVinfection,the icsA/virG fragment of Shigella-based prokaryotic expression plasmid pHS3199 was *** type 16 L1(HPV16L1)gene was inserted into plasmid pHS3199 to form the pHS3199-HPV16L1construct,and pHS3199-HPV16L1 was electroporated into a live attenuated Shigella strain *** analysis showed that HPV16L1 could be expressed stably in the recombinant strain *** test results were negative,which showed that the sh42-HPV16L1 lost ***,the at-tenuated recombinant strain partially maintained the invasive property as indicated by the HeLa cell *** IgG,IgA antibody against HPV16L1 virus-like particles(VLPs)were detected in the sera,intestinal lavage and vaginal lavage from animals immunized by *** number of antibody-secreting cells in the spleen and draining lymph nodes were increased significantly compared with the *** from immunized animals inhibited murine hemagglutination induced by HPV16L1 VLPs,whichindicated that the candidate vaccine could stimulate an efficient immune response in guinea pig's *** 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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PRELIMINARY STUDY OF A NOVEL human papillomavirus type 16 L1/E6-E7 CHIMERIC RECOMBINANT DNA VACCINE

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Journal of Pharmaceutical Analysis 2004年第1期16卷 45-49页

作者：郑瑾马军张福萍杨筱凤董小平司履生王一理 Institute of Cancer Research

Objective Preparations of HPV16 L1/E6 and L1/E7 prophylactic and therapeutic DNA vaccines. Methods The nucleotides within HPV16 E6 and E7 genes, which are responsible for viral transforming activity, were mutated by mage primer site-directed mutagenesis method. The correctly mutated E6 and E7 fragments were separately cloned into an eukaryotic expression vector pVAX1, together with HPV16 L1 gene, generating chimeric recombinants plasmids 1MpVAX1-L1E6, 2MpVAX1-L1E6, 1MpVAX1-L1E7, 2MpVAX1-L1E7 and 3MpVAX1-L1E7. CHO cells were transiently transfected with the individual DNA vaccines by calcium phosphate method. Target protein expressions in the extracts of the transfected cell lines were measured by ELISA and immunohistochemistry, with HPV16 L1 and E6 specific monoclonal antibodies. Results ELISA assays showed the P/N ratios in the cell extracts transfected with L1E6 and L1E7 plasmids were more than 2.1. Immunohistochemistry revealed brownish precipitant signal in cytoplasm and nuclei of the transfected cells. Conclusion Successful constructions of prophylactic and therapeutic DNA vaccine plasmids lay solid foundation for future animal experiment and clinical trial.

关键词： human papillomavirus type 16 DNA vaccine site-direct mutation

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Enhanced immunization after intranasal coadministration of Escherichia coli heat-labile enterotoxin B subunit and human papillomavirus 16-L1 DNA vaccine

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Chinese Medical Journal 2006年第5期119卷 408-411页

作者： WANG Jing ZHAO Chang-an WANG Kai ZHENG Jin WANG Yi-li SI Lü-sheng Key Laboratory of Biomedical Information Engineering of Education Ministry Institute for Cancer Research School of Life Science ＆ Technology Xi＇an Jiaotong University Xi＇an 710061 China

human papillomavirus （HPV）, mainly types 16 and 18, are the most important initiating agents of cervical cancer. Prevention of high-risk HPV infections is a potentially effective approach to control HPV associated c... 详细信息

关键词： Escherichia coil heat-labile enterotoxin B subunit human papillomavirus type 16 DNA vaccines

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Cloning of HPV16 E2 Gene from a Biopsied Cervical Cancer Sample

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Journal of Reproduction and Contraception 2002年第3期13卷 140-145页

作者： Yan-eGAO JuZHANG Tian-baoSONG Xiao-junYAN DepartmentofHistologyandEmbryology SchoolofMedicineXi＇anJiaotongUniversityXi＇an710061China InstituteofGeneDiagnosis FourthMilitaryMedicalUniversityXi＇an710032China DepartmentofHistologyandEmbryology SchoolofMedicineXi＇anJiaotongUniversityXi＇an710061China

To clone HPV16 E2 gene from a biopsied cervical cancer sample Materials & Methods HPV16 E2 gene was amplified from specimen derived from a HPV 16 positive patient, then cloned and sequenced. Results The full length of HPV 16 E2 gene was successfully cloned. In comparison with the prototype accepted by GenBank, six point mutations in HPV 16 E2 nucleotide acid sequence were identified. Of them, three were missense, and one was in the overlapping E4 gene and was synonymous to E4. Conclusion HPV16 E2 gene was successfully cloned, and some nucleotide acids in its sequence were different from the prototype.

关键词： human papillomavirus type 16 E2 gene cloning

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