Discovery and Reconstitution of Cross-Reactive Epitope Targets for H5 and H9 Avian Influenza
Exotic and Emerging Avian Viral Diseases Research Unit
2012 Annual Report
1a.Objectives (from AD-416):
Identify and map highly conserved B-cell epitopes common to surface of H5 and H9 avian influenza (AI) and to test their suitability as homotypic or universal vaccine components.
1b.Approach (from AD-416):
In this study, recombinant epitope mapping will be accomplished using sequence analysis of hemagglutinin genes of H5 and H9 avian influenza virus, and epitopes generated in comprehensive conformer-libraries to be displayed on filamentous bacteriophages. These epitopes will be selected for high evolutionary conservation and designed to elicit cross neutralization against diverse H5 and H9 avian influenza viruses. Most promising H5 and H9 epitopes will be used to immunize chickens and examine for ability to protect from H5 and H9 avian influenza virus challenge, respectively.
This project is related to objective 5 of this in-house project: Develop new vaccine platforms designed to control and prevent avian influenza virus outbreaks.
In FY2012, ten hens were vaccinated with 0.5ml dose of inactivated oil emulsion vaccine prepared with the AIV strain A/ck/Egypt/CLEVB-HK213 (9402NAMRU3)/2007 – H5N1. Three weeks after the first dose, the hens were boosted with a second vaccine dose, and a week after the booster the eggs were collected twice a day, individually labeled, and stored at 4C until testing. After testing, one hen with higher antibody titer by HI test was selected, and the IgY was purified using a commercial kit Pierce® Chicken IgY Purification Kit. The purified IgY was assayed for concentration, and shipped to Cooperator for use in the project.