详细记录  
题名:Heterologous viral RNA export elements improve expression of severe acute respiratory syndrome (SARS) coronavirus spike protein and protective efficacy of DNA vaccines against SARS
作者:Beno?t Callendret, Valrie Lorin, Pierre Charneau, Philippe Marianneau, Hugues Contamin, Jean-Michel Betton, Sylvie van der Werf and Nicolas Escriou
来源:Virology[IF=3.08], In Press, Corrected Proof, Available online 28 February 2007,
URL :http://dx.doi.org/10.1016/j.virol.2007.01.012
日期:070415
摘要: Beno?t Callendreta, , Valrie Lorina, , Pierre Charneaub, , Philippe Marianneaud, , Hugues Contamind, 1, , Jean-Michel Bettonc, , Sylvie van der Werfa, and Nicolas Escrioua, ,

aUnit de Gntique Molculaire des Virus Respiratoires, URA CNRS 1966, EA 302 Universit Paris 7, France
bGroupe 5 ans de Virologie Molculaire et de Vectorologie, France
cUnit de Biochimie Structurale, URA CNRS 2185, Institut Pasteur, 25 rue du Dr. Roux, 75724 PARIS Cedex 15, France
dUnit de Biologie des Infections Virales Emergentes, Institut Pasteur, IFR 128 BioSciences Lyon-Gerland, 21 avenue Tony Garnier, 69365 Lyon Cedex 07, France

Received 10 October 2006; revised 19 December 2006; accepted 15 January 2007. Available online 28 February 2007.

Summary

The SARS-CoV spike glycoprotein (S) is the main target of the protective immune response in humans and animal models of SARS. Here, we demonstrated that efficient expression of S from the wild-type spike gene in cultured cells required the use of improved plasmid vectors containing donor and acceptor splice sites, as well as heterologous viral RNA export elements, such as the CTE of Mazon-Pfizer monkey virus or the PRE of Woodchuck hepatitis virus (WPRE). The presence of both splice sites and WPRE markedly improved the immunogenicity of S-based DNA vaccines against SARS. Upon immunization of mice with low doses (2 g) of naked DNA, only intron and WPRE-containing vectors could induce neutralizing anti-S antibodies and provide protection against challenge with SARS-CoV. Our observations are likely to be useful for the construction of plasmid and viral vectors designed for optimal expression of intronless genes derived from cytoplasmic RNA viruses. 31.

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