title: An infected chicken kidney cell co-culture ELISpot for enhanced detection of T cell responses to avian influenza and vaccination.
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.
authors.affiliation: The Pirbright Institute, Compton Laboratory, Compton, Newbury, Berkshire RG20 7NN, UK.
authors.forename: Raul
authors.initials: R
authors.lastname: Ruiz-Hernandez
authors.affiliation: The Pirbright Institute, Compton Laboratory, Compton, Newbury, Berkshire RG20 7NN, UK.
authors.forename: Marylene
authors.initials: M
authors.lastname: Peroval
authors.affiliation: University of Oxford, Department of Zoology, South Parks Road, Oxford OX1 3PS, UK.
authors.forename: Amy
authors.initials: A
authors.lastname: Boyd
authors.affiliation: The Pirbright Institute, Compton Laboratory, Compton, Newbury, Berkshire RG20 7NN, UK.
authors.forename: Devanand
authors.initials: D
authors.lastname: Balkissoon
authors.affiliation: The Pirbright Institute, Compton Laboratory, Compton, Newbury, Berkshire RG20 7NN, UK.
authors.forename: Karen
authors.initials: K
authors.lastname: Staines
authors.affiliation: University of Oxford, Department of Zoology, South Parks Road, Oxford OX1 3PS, UK.
authors.forename: Adrian
authors.initials: A
authors.lastname: Smith
authors.affiliation: The Pirbright Institute, Compton Laboratory, Compton, Newbury, Berkshire RG20 7NN, UK. Electronic address: colin.butter@avianimmunology.org.
authors.forename: Colin
authors.initials: C
authors.lastname: Butter
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D016827:CD8 Antigens / Q000276:immunology; D002460:Cell Line; D002645:Chickens / Q000276:immunology* / Q000821:virology; D018920:Coculture Techniques / Q000379:methods; D004285:Dogs; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods; D005585:Influenza in Birds / Q000276:immunology* / Q000821:virology; D007371:Interferon-gamma / Q000276:immunology; D007668:Kidney / Q000276:immunology* / Q000821:virology; D061985:Madin Darby Canine Kidney Cells; D013601:T-Lymphocytes / Q000276:immunology* / Q000821:virology; D014611:Vaccination / Q000379:methods
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D016827:CD8 Antigens; D007371:Interferon-gamma
keywords: Adaptive antiviral responses; Antigen presentation; Avian influenza; CD8 T cell; IFNγ ELISpot