An enzyme-linked procedure for the detection and estimation of surface receptors on cells.
An enzyme immunoassay procedure has been developed for the visualization and estimation of lymphocyte surface receptors. beta-Galactosidase (beta-gal'ase) was covalently linked to sheep anti-rabbit immunoglobulin (SARIg), to ovalbumin (OA), and to adenosine (A). Exposure of rabbit peripheral lymphocytes to SARIg-beta-gal'ase and subsequent incubation with the fluorogenic substrate fluorescein-beta-digalactopyranoside allowed visualization of B cells by fluorescence microscopy. Treatment with each of the above beta-gal'ase conjugates and incubation with another fluorogenic substrate, 4-methyl-beta-D-umbelliferylgalactopyranoside, made possible the estimation of the various receptors by spectro fluorimetry. Procedures used for enrichment of T and B cell populations could be monitored. Among the findings was that immunoglobulin-bearing cells (presumably B cells) formed rosettes with sheep erythrocytes. The quantitative procedure was used to study lymphocyte population changes during immunization with an A-O A conjugate. An increase in the binding of A-beta-gal'ase, SARIg-beta-gal'ase, and OA-beta-gal'ase by peripheral lymphocytes occurred repeatedly 6 to 8 days after immunization but lasted only a few days despite a continued high circulating antibody titer. These results are consistent with the possibility that immunization induces migration of lymphocytes into the circulation from bone marrow and/or thymus.
D000241:Adenosine / Q000276:immunology; D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D001402:B-Lymphocytes / Q000276:immunology*; D001667:Binding, Competitive; D005455:Fluorescent Antibody Technique; D005696:Galactosidases / Q000378:metabolism; D007114:Immunization; D007118:Immunoassay / Q000379:methods; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000276:immunology; D010047:Ovalbumin / Q000276:immunology; D011817:Rabbits; D011947:Receptors, Antigen, B-Cell / Q000032:analysis*; D012756:Sheep; D013601:T-Lymphocytes / Q000276:immunology*
D000888:Antibodies, Anti-Idiotypic; D011947:Receptors, Antigen, B-Cell; D010047:Ovalbumin; D005696:Galactosidases; D000241:Adenosine