Abstract
Duck is susceptible to many pathogens, such as duck hepatitis virus, duck enteritis virus (DEV), duck tembusu virus, H5N1 highly pathogenic avian influenza virus (HPAIV) in particular. With the significant role of duck in the evolution of H5N1 HPAIV, control and eradication of H5N1 HPAIV in duck through vaccine immunization is considered an effective method in minimizing the threat of a pandemic outbreak. Consequently, a practical strategy to construct a vaccine against these pathogens should be determined. In this study, the DEV was examined as a candidate vaccine vector to deliver the hemagglutinin (HA) gene of H5N1, and its potential as a polyvalent vaccine was evaluated. A modified mini-F vector was inserted into the gB and UL26 gene junction of the attenuated DEV vaccine strain C-KCE genome to generate an infectious bacterial artificial chromosome (BAC) of C-KCE (vBAC-C-KCE). The HA gene of A/duck/Hubei/xn/2007 (H5N1) was inserted into the C-KCE genome via the mating-assisted genetically integrated cloning (MAGIC) to generate the recombinant vector pBAC-C-KCE-HA. A bivalent vaccine C-KCE-HA was developed by eliminating the BAC backbone. Ducks immunized with C-KCE-HA induced both the cross-reactive antibodies and T cell response against H5. Moreover, C-KCE-HA-immunized ducks provided rapid and long-lasting protection against homologous and heterologous HPAIV H5N1 and DEV clinical signs, death, and primary viral replication. In conclusion, our BAC-C-KCE is a promising platform for developing a polyvalent live attenuated vaccine.Electronic supplementary materialThe online version of this article (doi:10.1186/s13567-015-0174-3) contains supplementary material, which is available to authorized users.
Highlights
Ducks are considered one of the most important waterfowl for its various usages in different aspects
Establishing a full-length C-KCE clone harboring mini-F plasmid sequences Establishing a full-length C-KCE clone in E. coli first requires the insertion of a bacterial artificial chromosome (BAC) vector into the viral genome (Figure 1A)
The BAC vector was inserted into a large junction of the gB and UL26 genes in the C-KCE genome (Figure 1B)
Summary
Ducks are considered one of the most important waterfowl for its various usages in different aspects. In China and southeast Asia, duck farming is a traditional agribusiness for nourishment, and critical for habiliment. This traditional business is seriously threatened by numerous pathogens, such as avian influenza virus (AIV), duck hepatitis virus, duck enteritis virus (DEV), and duck tembusu virus [1,2]. It is currently known that almost all the subtypes can be isolated from waterfowl with the. Under these circumstances, vaccination, as an adjunct for improving bio-security and stamping-out policies, contributes to protecting ducks against AIV infection [9].
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