[Construction and immunogenicity analysis of recombinant replication-defective human adenovirus type 5 bearing the porcine circovirus type 2 Cap protein gene].
Bing Du Xue Bao
; 29(1): 26-31, 2013 Jan.
Article
em Zh
| MEDLINE
| ID: mdl-23547376
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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes
/
Adenovírus Humanos
/
Circovirus
/
Vírus Defeituosos
/
Proteínas do Capsídeo
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Bing Du Xue Bao
Assunto da revista:
VIROLOGIA
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
China