Construction of recombinant fowlpox virus coexpressing HA gene from H5N1 avian influenza virus and chicken interleukin-2 gene and assessment of its protective efficacy.
- Author:
Shui-Li YUN
1
;
Wei ZHANG
;
Wu-Ji LIU
;
Xiao-Rong ZHANG
;
Su-Juan CHEN
;
Yan-Tao WU
;
Da-Xin PENG
;
Xiu-Fan LIU
Author Information
1. Key Laboratory of Animal Infectious Diseases, Ministry of Agriculture, Yangzhou University, Yangzhou 225009, China. yunshuili2008@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Animals;
Cells, Cultured;
Chick Embryo;
Chickens;
Fowlpox virus;
genetics;
metabolism;
Gene Expression;
Genetic Engineering;
Genetic Vectors;
genetics;
metabolism;
Hemagglutinins;
genetics;
immunology;
Influenza A Virus, H5N1 Subtype;
genetics;
immunology;
Influenza in Birds;
immunology;
virology;
Interleukin-2;
genetics;
immunology;
Random Allocation
- From:
Chinese Journal of Virology
2009;25(6):430-436
- CountryChina
- Language:Chinese
-
Abstract:
The hemagglutinin (HA) gene from H5N1 avian influenza virus and the chicken interleukin 2 (chiIL-2) gene were inserted into a expressing vector p12LS to construct a recombinant transferring vector p12LSH5AIL2, in which HA gene under the control of the promoter Ps was in inverse tandem connection with the chiIL-2 gene under the control of the promoter PE/L. The p12LSH5AIL2 was then used to transfect the chicken embryo fibroblasts (CEF) pre-infected with a wild-type fowlpox virus 282E4 strain, to generate a recombinant fowlpox virus coexpressing the inserted HA and chiIL2 genes (rFPV-H5AIL2). The rFPV-H5AIL2 was obtained and purified by blue plaque screening on the CEF. The in vitro expression of HA gene by rFPV-H5AIL2 was detected in the recombinant fowlpox virus-infected CEFs with an indirect immunofluorescence assay, and the expression of the chiIL2 gene by rFPV-H5AIL2 was confirmed by detection of the chiIL2 mRNA by RT-PCR and by detection of chiIL2 by the indirect immunofluorescence assay. Experiments on SPF and commercial chickens demonstrated that the titer for HI antibodies induced by the rFPV-H5AIL2 was significantly higher than that by the rFPV-HA. The group immunized with the rFPV-H5AIL2 exhibited the similar ratios of protective efficacy and virus shedding as the group immunized with the rFPV-HA in SPF chicken. However, in commercial chicken, the group immunized with the rFPV-H5AIL2 generated significantly higher protection against H5N1 avian influenza virus challenge and lower virus shedding than the group immunized with the rFPV-HA. This study paved the way for further development of a new AIV recombinant vaccine.
