Use of enzyme microassay to detect eosinophil potentiating activity of cytokines in sheep |
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Authors: | Lesley M. Stevenson Douglas G. Jones |
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Affiliation: | Moredun Research Institute, Edinburgh, UK |
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Abstract: | Abstract Microplate assays for eosinophil peroxidase (EPO) and arylsulphatase (EAS) have been established as indices of eosinophil survival/proliferation in sheep bone marrow cell (SBMC) cultures. Cell specificity was confirmed using density-fractionated and differentially depleted SBMC populations. Several recombinant cytokines including interleukins 3 (IL-3) and 5 (IL-5), and granulocyte macrophage-colony stimulating factor (GM-CSF), but not macrophage-CSF (M-CSF), had demonstrable eosinophil-potentiating activity on the basis of enhanced EPO and EAS activities in treated, compared with untreated, SBMC cultures. Effects of IL-5 were abrogated in the presence of a specific monoclonal anti-IL-5 antibody. The results indicate that measurement of EPO and EAS in cultured SBMC offers a simple and effective method for detecting eosinophil potentiating activity in the ovine. |
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Keywords: | Peroxidase/arylsulphatase microassay Eosinophil potentiating activity Cytokine Sheep bone marrow |
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