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| Construction and Immunogenicity of a Recombinant Adenovirus Expressing NSP4 of Group A Rotavirus |
| XIE Li1,2, WANG Xiao-nan1, LI Yang1, YAN Min1, CAO Yin1, LI Hong-jun1, SUN Mao-sheng1 |
1. Institute of Medical Biology, Chinese Academy of Medical Sciences/Peking Union Medical College; Yunnan Key laboratory of Vaccine Research & Development on Severe Infections Diseases, Kunming 650118, China;
2. Medical School of Kunming University, Kunming 650214, China |
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Abstract Objective:To construct a recombinant adenovirus expressing the non-structure protein 4(NSP4) of A group rotavirus, and evaluate its immune response effects. Methods: NSP4 gene from wildtype rotavirus was cloned into a shuttle plasmid (pshuttle-CMV). The pshuttle-NSP4 is transformed into E.coli BJ5183 to make a homologous recombination with adenovirus skeleton genome(pAdeasy). And the recombinant plasmid pAd-NSP4 is transfected into Ad-293 cell to produce mature adenovirus particles. ICR mice are vaccinated with this recombinant adenovirus by intramuscular and intranasal administration. Results: The recombinant adenovirus titer rise to 108.25CCID50/ml after 3 time passaged. The transcription and expression of NSP4 can be determined by RT-PCR and immunofluorescene assay in vitro. In intramuscular and intranasal administration group, ELISA determines specific serum antibody mean titer at 1:320 and 1:1436.8 level; neutralizing antibody titer at 1:45.3 and 1:71.8. Conclusion: The recombinant adenovirus containing rotavirus NSP4 gene can induce effective immune response against rotavirus. Intranasal administration produce a better protective efficacy than intramuscular in mice. It may be a good candidate to develop a novel engineering vaccine in the future.
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Received: 03 December 2013
Published: 25 February 2014
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