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United States Department of Agriculture

Agricultural Research Service

Title: Development of Hemagglutinin Subtype Specific Reference Antisera by DNA Vaccination of Chickens

Authors
item Lee, Chang-Won - USDA-ARS-SEPRL-STUDENT
item Senne, Dennis - NVSL - AMES,IA
item Suarez, David

Submitted to: Avian Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 14, 2002
Publication Date: September 1, 2003
Citation: Lee, C., Senne, D., Suarez, D.L. 2003. Development of hemagglutinin subtype specific reference antisera by DNA vaccination of chickens. Avian Diseases 47:1051-1056, 2003.

Interpretive Summary: In the current nomenclature for influenza A viruses, 15 hemagglutinin (H) subtypes have been described. All 15 H subtypes of influenza virus are found in aquatic birds, which serve as the reservoir of all influenza A viruses. The hemagglutination-inhibition test using reference antisera is a standard method to confirm and subtype influenza A viruses. For this test, preparation of antisera of good quality is essential. Most commonly reference antisera is made by injecting chickens with live or killed whole virus preparations to stimulate an antibody response. Previously, we showed that intramuscular vaccination of chickens with the DNA (pCI-neo plasmid), expressing the influenza protein, could stimulate a measurable and protective antibody response. In this study, we extended our previous experiment to prepare reference antisera against several H subtypes by DNA vaccination. Subtype specificity of the antisera prepared by DNA vaccination were comparable or better than the antisera prepared by traditional method using whole virus vaccination. Preparation of reference antisera by DNA vaccination holds good promise because it is safe and allows for the production of H specific antibodies without producing antibodies specific to other influenza viral proteins.

Technical Abstract: We previously showed that intramuscular vaccination of chickens with the eukaryotic expression vector (EEV), expressing the influenza H5 hemagglutinin (H) protein, can stimulate a measurable and protective antibody response. Based on these results, we cloned other H genes from Eurasian H5, North American and Eurasian H7, and H15 influenza viruses into othe EEV for use in vaccination of chickens to produce reference antibodies for diagnostic purposes, such as the hemagglutination inhibition (HI) test. Three week old SPF chickens were vaccinated with 100ug of EEV mixed with a cationic lipid by intramuscular injection. Then, the birds were boostered twice at monthly intervals after the original vaccination. Measurable antibody titers were present for most birds after one month, and generally increased after each boost. To examine the cross reactivity of the sera with other subtypes, HI test was conducted with antigens prepared from 15 subtypes of influenza virus. Subtype specificity of the antisera prepared by DNA vaccination were comparable or better than the antisera prepared by traditional method using whole virus vaccination. Preparation of reference antisera by DNA vaccination holds good promise because it is safe and allows for the production of H specific antibodies without producing antibodies specific to other influenza viral proteins.

Last Modified: 8/2/2014
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