Construction and immunogenicity analysis of recombinant replication-defective human adenovirus type 5 bearing the porcine circovirus type 2 Cap protein gene.
- Author:
Ting GONG
1
;
Shou-Feng ZHANG
;
Ye LIU
;
Cheng-Long SUN
;
Yang YANG
;
Qi CHEN
;
Fang QIAN
;
Bo-Hao LIU
;
Rong-Liang HU
Author Information
1. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China. gongting2010@126.com
- Publication Type:Journal Article
- MeSH:
Adenoviruses, Human;
genetics;
Animals;
Antibodies, Viral;
blood;
Capsid Proteins;
genetics;
immunology;
Circovirus;
immunology;
Defective Viruses;
genetics;
HEK293 Cells;
Humans;
Mice;
Recombinant Proteins;
biosynthesis;
immunology;
Virus Replication
- From:
Chinese Journal of Virology
2013;29(1):26-31
- CountryChina
- Language:Chinese
-
Abstract:
To construct a recombinant replication-defective human adenovirus type 5 expressing Cap protein of PCV2 and test the immunological efficacy in mice. In this study, the recombinant replication-defective human adenovirus type 5, named as rAd5-Cap (wt-rAd5), was constructed through homologous recombination internally in the HEK293AD cells after co-transfection of the Pac I-linearized backbone plasmid and the shuttle plasmid pacAd5CMV-Cap containing the open reading frame (ORF2) of the porcine circovirus type 2 (PCV2) cap protein or pacAd5CMV without inserted fragment. Furthermore, the rAd5-Cap could induce the expression of PCV2 cap protein in the HEK293AD cells with high efficacy evaluated by the RT-PCR and indirect immunofluorescence assay (IFA). The virus titer of rAd5-Cap could reach up to 10(8.5) TCID50/mL similarly to that of wt-rAd5, indicating that there was little affect on the virus proliferation after the insertion of PCV2 cap protein gene. The humeral immune responses could be activated and detected 14 days after the inoculation of the mice with 10(7) TCID50 rAd5-Cap intramuscularly, and constantly in crease in another 14 days. These molecular biological and animal experiments results demonstrated that the PCV2 cap protein could be efficiently expressed by the recombinant adenovirus rAd5-Cap in eukaryotic cells and induce robust immune responses in mice, which laid a good foundation for the development of new type vaccine against porcine circovirus.
