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| Development and Characterization of Integration-deficient Lentiviral Vector and HCV Recombinant Pseudotyped Lentiviral Particles |
| GUAN Jie1,2, DENG Yao1, WEN Bo1, CHEN Hong1, WANG Wen1, TAN Wen-Jie1 |
1. Key Laboratory of Medical Virology, Ministry of Health; National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206, China;
2. Central Laboratory, Meitan General Hospital, Beijing 100028, China |
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Abstract In order to improve the safety of lentiviral vector and improve the preparation of novel hepatitis C virus (HCV) particle vaccine,the traditional three plasmid-base lentiviral system was modificated: the packaging plasmid pHR’CMV Δ R8.2 was transformed into the integration-deficient packaging plasmid pCMVΔR8.2D64E, and confirmed the integration-defective properties in vitro. The reporter gene (GFP) in transfer plasmid was replaced by the HCV non-structural gene NS3. HCV envelope plasmid with different subtypes (from three representative subtypes 1a or 1b or 2a) combined with the above two modified lentiviral plasmids were cotransfected 293FT cells. HCV integration-defective pseudotyped lentiviral particle (IDLVpp) vaccines carrying HCV NS3 were packaged. The HCV IDLVpp were identified by Western blot, and the particle structure was observed by electron microscope. The expression of transgenic HCV NS3 in Huh7 cells infected by HCV IDLVpp was also observed. So a scientific and experimental basis for novel effective HCV vaccines development and applications of HCV IDLVpp vaccines was provided.
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Received: 29 January 2013
Published: 25 June 2013
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