Cloning and expression of the extra-cellular part of the alpha chain of the equine high-affinity IgE receptor and its use in the detection of IgE.
The high-affinity receptor for IgE (FcepsilonRI) plays a central role in IgE-mediated allergic reactions. Cross-linking of FcepsilonRI by IgE-antigen complexes results in the activation of mast cells and basophils and is thought to contribute to the immunopathology of Heaves, a chronic obstructive pulmonary disease of horses. Recombinant protein corresponding to the extra-cellular portion of the FcepsilonRI alpha subunit, cloned and sequenced previously, was expressed using both mammalian cells and insect cells. The yield of expressed protein was considerably greater using insect cells and the baculovirus expression system. The recombinant proteins differed in size between the two systems, presumably due to differences in the extent of glycosylation. However, recombinant protein from both cell systems bound equine IgE present in bronchoalveolar lavage fluid from horses with Heaves. These results suggest that the recombinant extra-cellular part of FcepsilonRI should be a useful tool with which to study equine IgE responses.
McAleese
S M
SM
Department of Veterinary Clinical Studies, University of Edinburgh, R(D)SVS, Easter Bush Veterinary Centre, Roslin, Midlothian EH25 9RG, UK.
Brown
J K
JK
Macrae
A I
AI
Mackellar
A
A
Huntley
J F
JF
Miller
H R P
HR
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western / Q000662:veterinary; D001992:Bronchoalveolar Lavage Fluid / Q000276:immunology; D019556:COS Cells; D002522:Chlorocebus aethiops; D006734:Horse Diseases / Q000175:diagnosis / Q000276:immunology*; D006736:Horses; D007073:Immunoglobulin E / Q000276:immunology*; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D029424:Pulmonary Disease, Chronic Obstructive / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D017455:Receptors, IgE / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D010446:Peptide Fragments; D017455:Receptors, IgE; D011994:Recombinant Proteins; D007073:Immunoglobulin E