Identification and Expression of a Gene Encoding an Epitope that Induces Hemagglutination Inhibition Antibody to Avibacterium paragallinarum Serovar A

生物 表位 单克隆抗体 血凝试验 重组DNA 分子生物学 病毒学 免疫原性 抗体 免疫印迹 基因 遗传学 效价 病毒
作者
Taichi Noro,Kazuhiko Yaguchi,Katsuhiko Amimoto,Eiji Oishi
出处
期刊:Avian Diseases [BioOne (American Association of Avian Pathologists)]
被引量:15
标识
DOI:10.1637/0005-2086(2007)051[0084:iaeoag]2.0.co;2
摘要

The aims of this study were the identification, cloning, and expression of a genetic region encoding an epitope that induces hemagglutination inhibition (HI) antibody against Avibacterium paragallinarum serovar A and an evaluation of the recombinant protein for immunogenicity in chickens. Although two monoclonal antibodies (MAbs) with HI activity, designated S24-951 and S7-1716-5C, were generated in this study, no reactive proteins with both MAbs were identified by Western blot analysis. A gene fragment of 5157 bp, designated hpa5. 1, was cloned from genomic DNA, and a recombinant protein expressed by hpa5.1, designated HPA5.1, reacted with both MAbs on dot-blot analysis. HPA5.1 showed no hemagglutinating activity, but significantly absorbed HI antibodies in the chicken immune serum. Analysis using a series of deletion mutants prepared from hpa5.1 indicated that a 4.8 kbp gene in hpa5.1 is essential for the expression epitope recognized by MAb S24-951. In addition, chickens immunized once with HPA5.1 showed a high protection rate with sufficient HI antibody titers against challenge exposure with a virulent strain of A. paragallinarum serovar A strain 221. These results show that hpa5. I1 is responsible for the expression of an epitope that induces HI antibody, and HPA5.1 might be a candidate for the development of a new vaccine against avian infectious coryza caused by A. paragallinarum serovar A.
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