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

Autor: Marylene Y. Peroval, Karen Staines, Devanand Balkissoon, C. Butter, Adrian Smith, Raul Ruiz-Hernandez, Amy Boyd
Rok vydání: 2014
Předmět:
Fowlpox
Enzyme-Linked Immunospot Assay
T-Lymphocytes
Avian influenza
medicine.disease_cause
Antibodies
Viral
Kidney
Madin Darby Canine Kidney Cells
0302 clinical medicine
Cytotoxic T cell
Immunology and Allergy
Antigens
Viral
0303 health sciences
ELISPOT
Vaccination
CKC
Primary chicken kidney cells
3. Good health
rFPV
Recombinant Fowlpox virus
medicine.anatomical_structure
CTL
Cytotoxic T lymphocytes
Research Paper
T cell
CD8 Antigens
Immunology
MVA
Modified Vaccinia virus Ankara
Biology
Virus
Cell Line
03 medical and health sciences
Interferon-gamma
Dogs
Antigen
APC
Antigen presenting cells
Adaptive antiviral responses
IFNγ ELISpot
medicine
Animals
Antigen presentation
030304 developmental biology
MHC
Major histocompatibility complex
ChIFNγ
Chicken interferon-gamma
medicine.disease
Virology
Influenza A virus subtype H5N1
Coculture Techniques
CD8 T cell
Influenza in Birds
Chickens
030215 immunology
Zdroj: Journal of Immunological Methods
ISSN: 1872-7905
Popis: 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.
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.
Databáze: OpenAIRE
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