title: Efficacy of a high-yield attenuated vaccine strain wholly derived from avian influenza viruses by use of reverse genetics.
abstract: The preparation of high-yield influenza H5N1 vaccine strains is challenging for researchers and manufacturers. Here, we used reverse genetics to generate a high-yield avian influenza vaccine strain based on a novel avian influenza virus. A high-yield attenuated recombinant H5N3 virus (rH5N3-DL) was prepared from the HA gene of A/Goose/Anhui/08 (H5N1), modified by deletion of the multiple basic amino acids at the cleavage site, the NA gene from A/Duck/Germany/1215/73 (H2N3), and the six internal genes from the high-yield A/Goose/Dalian/3/01 (H9N2) virus. rH5N3-DL grew to high HA titers (1:2048) in eggs, eight times those of the parental H5N1 virus, and four times higher than that of rH5N3-PR8 (six internal genes from the high-yield PR8). Infection tests demonstrated that rH5N3-DL was avirulent in chickens, chicken embryos, and mice. rH5N3-DL-vaccinated chickens were fully protected against the morbidity and mortality of a lethal challenge with homologous A/Goose/Anhui/08, but only 80% of chickens were protected after challenge with heterologous A/Goose/Guangdong/1/96. The N3 neuraminidase marker distinguishes rH5N3-DL-vaccinated from H5N1-infected animals. rH5N3-DL is thus a promising vaccine candidate to combat highly pathogenic avian influenza virus infections. The A/Goose/Dalian/3/01 virus could be a promising candidate as providing internal genes donors with high-yield properties in reverse-genetics system and might be applicable for future avian influenza vaccine development.
authors: Animal influenza Laboratory of Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of CAAS, Harbin 150001, PR China. liuming04@126.com
authors: Ming
authors: M
authors: Liu
authors: Chun-Guo
authors: CG
authors: Liu
authors: Yun
authors: Y
authors: Zhang
authors: Wei-Lin
authors: WL
authors: Shi
authors: Wei
authors: W
authors: Wang
authors: Yan-Yun
authors: YY
authors: Liu
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002645:Chickens / Q000276:immunology / Q000821:virology; D057809:HEK293 Cells; D006801:Humans; D009980:Influenza A virus / Q000235:genetics / Q000276:immunology; D007252:Influenza Vaccines / Q000235:genetics* / Q000276:immunology*; D005585:Influenza in Birds / Q000276:immunology* / Q000401:mortality / Q000517:prevention & control / Q000821:virology; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D059386:Reverse Genetics; D014613:Vaccines, Attenuated / Q000235:genetics / Q000276:immunology; D014779:Virus Replication
chemical_list: D000914:Antibodies, Viral; D007252:Influenza Vaccines; D014613:Vaccines, Attenuated