An infected chicken kidney cell co-culture ELISpot for enhanced detection of T cell responses to avian influenza and vaccination

Highlights

• Chickens infected with avian influenza developed IFNγ responses.

• The use of infected CKC in ELISpot overcomes limitations at detection of responses.

• This methods allows the quantification of influenza specific CD8 T cells.

• The use of recombinant virus to infect CKC can further define antigen specificity.

Abstract

A better understanding of the immune responses of chickens to the influenza virus is essential for the development of new strategies of vaccination and control. We have developed a method incorporating infected chicken kidney cells (CKC) in culture with splenocytes in an IFNγ ELISpot assay to enumerate ex vivo responses against influenza virus antigens. Splenocytes from birds challenged with influenza showed specific responses to the influenza virus, with responding cells being mainly CD8 positive. The utility of the assay was also demonstrated in the detection of an antigen specific enhancement of IFNγ producing cells from birds vaccinated with recombinant Fowlpox vectored influenza nucleoprotein and matrix protein.