DETECTION OF MOUSE ALLOANTIBODIES BY ROSETTING WITH PROTEIN A-COATED SHEEP RED BLOOD CELLS

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Abstract
Staphylococcal protein A has an affinity for the Fc portion of the Ig[immunoglobulin]G molecule of different species and can therefore be used to detect cell-bound Ig. Using this property, protein A coupled to sheep red blood cells via chromic chloride can detect alloantibodies to mouse H-2; Thy-1; Ly-1, 2, 4, 5, 6 and 7; and Ia [immune response associated] antigenic specificities bound to the surface of lymphocytes by the formation of rosettes. In comparison with other rosetting and cytotoxicity assays, the protein A assay shows a greater sensitivity than does cytotoxicity using spleen cells as the target, as does the sheep anti-mouse Ig rosetting assay, whereas cytotoxicity shows greater sensitivity with some antisera on thymocytes. The major advantages of the protein A assay are that constant low reproducible backgrounds are obtained, there is no need to remove surface Ig by capping prior to antiserum treatment and viable cells can be recovered.