Dose response effects of avian influenza (H7N7) vaccination of chickens: Serology, clinical protection and reduction of virus excretion

Knowledge of the relation between the antigen content of inactivated avian influenza (Al) vaccines, the serological response after vaccination and protection of vaccinated animals is important for the choice of optimal vaccines and vaccination regimes as well as for the assessment of criteria for the licensing of new Al-vaccines. We studied this relation in a dose response study using inactivated H7N7 avian influenza vaccines with varying antigen content. The serological response depended on the antigen content of the vaccines. Anti-Al antibodies were detected most frequently with ELISA, followed by the virus neutralisation test and the haemagglutination inhibition (HI) assay. Chickens with measurable HI-antibody titers, using homologous H7N7 antigen. were all protected against clinical disease after challenge with highly pathogenic A/chicken/Netherlands/621557/03 H7N7 virus. However, in these chickens high levels of virus could still be present on days 2-4 after challenge. The reduction of virus titers after challenge, depended on the antigen content of the vaccines as well as on the serum antibody titers. While 10 haemagglutinating units (HAU), equivalent to 0.8 mu g haemagglutinin (HA) protein, per vaccine dose was sufficient for prevention of clinical disease, 128 HAU (9 mu g HA) per dose was required for reduction of virus titers in all chickens to 10(3) egg-infectious dose 50% (EID50) or less. In order to reduce virus titers below 10(3) EID50 per swab a HI-antibody titer of 64 was required. After use of the vaccine with the highest antigen content, challenge still induced a booster of antibody titers which is indicative of replication of challenge virus