title: [Construction of recombinant fowlpox virus coexpressing HA gene from H5N1 avian influenza virus and chicken interleukin-2 gene and assessment of its protective efficacy].
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.
authors.affiliation: Key Laboratory of Animal Infectious Diseases, Ministry of Agriculture, Yangzhou University, Yangzhou 225009, China. yunshuili2008@yahoo.com.cn
authors.forename: Shui-Li
authors.initials: SL
authors.lastname: Yun
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Wu-Ji
authors.initials: WJ
authors.lastname: Liu
authors.forename: Xiao-Rong
authors.initials: XR
authors.lastname: Zhang
authors.forename: Su-Juan
authors.initials: SJ
authors.lastname: Chen
authors.forename: Yan-Tao
authors.initials: YT
authors.lastname: Wu
authors.forename: Da-Xin
authors.initials: DX
authors.lastname: Peng
authors.forename: Xiu-Fan
authors.initials: XF
authors.lastname: Liu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D005587:Fowlpox virus / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006388:Hemagglutinins / Q000235:genetics / Q000276:immunology*; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000276:immunology*; D005585:Influenza in Birds / Q000276:immunology* / Q000821:virology; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology*; D011897:Random Allocation
chemical_list: D006388:Hemagglutinins; D007376:Interleukin-2