Construction of a pigeonpox virus recombinant: expression of the Newcastle disease virus (NDV) fusion glycoprotein and protection of chickens against NDV challenge

Abstract

Summary A pigeonpox transfer plasmid was constructed by cloning a 2.5 kb DNA fragment containing the viral thymidine kinase (TK) gene in the psp65 plasmid. The vaccinia virus P11K promoter followed by the NDV fusion (F) gene was inserted in the TK gene. The F gene was transferred to the viral genome by homologous recombination in pigeonpox virus infected CEF cells, transfected with the recombinant plasmid. Recombinant viruses were selected with BUdR and screened for their ability to induce fusion between adjacent cells. Because of the unexpected growth advantage of the TK+ WT over the TK− recombinants, viral purification was needed to obtain stable recombinants expressing a glycosylated and cleaved F protein. Vaccination of chickens by the follicular method induced high anti-F antibody titers and good protection against challenge with the virulent Italian NDV strain. Half of the oculonasal vaccinated chickens showed anti F antibodies and also half of them were protected. Although protection seems to be correlated with antibody titers, no neutralizing antibodies were found.