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

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 enumer...

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Bibliographic Details
Published in:Journal of immunological methods 2015-01, Vol.416, p.40-48
Main Authors: Ruiz-Hernandez, Raul, Peroval, Marylene, Boyd, Amy, Balkissoon, Devanand, Staines, Karen, Smith, Adrian, Butter, Colin
Format: Article
Language:English
Subjects:
Adaptive antiviral responses
Animals
Antibodies, Viral - immunology
Antigen presentation
Antigens, Viral - immunology
Avian influenza
CD8 Antigens - immunology
CD8 T cell
Cell Line
Chickens - immunology
Chickens - virology
Coculture Techniques - methods
Dogs
Enzyme-Linked Immunospot Assay - methods
IFNγ ELISpot
Influenza in Birds - immunology
Influenza in Birds - virology
Interferon-gamma - immunology
Kidney - immunology
Kidney - virology
Madin Darby Canine Kidney Cells
Research Paper
T-Lymphocytes - immunology
T-Lymphocytes - virology
Vaccination - methods
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Summary: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. •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.
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2014.10.012