Use of enzyme microassay to detect eosinophil potentiating activity of cytokines in sheep.
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.
Stevenson
L M
LM
Moredun Research Institute, Edinburgh, UK.
Jones
D G
DG
D000818:Animals; D001192:Arylsulfatases / Q000378:metabolism*; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology*; D047209:Eosinophil Peroxidase; D004804:Eosinophils / Q000201:enzymology / Q000276:immunology*; D010544:Peroxidases / Q000378:metabolism*; D012756:Sheep
D016207:Cytokines; D047209:Eosinophil Peroxidase; D010544:Peroxidases; D001192:Arylsulfatases