Abstract
Aim:
To develop a quantitative ELISA by measuring interferon (IFN-gamma) of equine lymphocytes.
Methods:
Sandwich ELISA for equine IFN-gamma was developed using mAb A5 as a capture antibody and biotinylated mAb SB10 as a detection antibody.
Results:
The detection limit of the sandwich ELISA for equine IFN-gamma was 1 microg/L and did not show cross-reactivity with recombinant equine IL-18. Equine IFN-gamma was detected by ELISA in culture medium of the peripheral blood mononuclear cells (PBMCs) stimulated with ConA or PMA/Ionomycin.
Conclusion:
This method can be used to help understand the role of this cytokine in various equine diseases and develop specific cell-mediated immunity assay.
MeSH terms
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Animals
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Antibodies, Monoclonal* / isolation & purification
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Biological Assay
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Cytokines / immunology
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Cytokines / isolation & purification*
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Enzyme-Linked Immunosorbent Assay / methods*
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Horses
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Interferon-gamma / analysis
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Interferon-gamma / immunology
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Interferon-gamma / isolation & purification*
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Interleukin-18 / immunology
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Interleukin-18 / isolation & purification*
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Ionomycin / chemistry
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Limit of Detection*
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Lymphocyte Activation
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Neutralization Tests
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Recombinant Proteins
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Sepharose / analogs & derivatives
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Sepharose / chemistry
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T-Lymphocytes, Regulatory
Substances
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Antibodies, Monoclonal
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Cytokines
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Interleukin-18
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Recombinant Proteins
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concanavalin A-sepharose
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Ionomycin
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Interferon-gamma
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Sepharose