Flow sorting of antigen-binding B cell subsets.
Antigen binding was used as a probe in the definition of functional B cell heterogeneity. Unprimed, anti-Thy 1 and complement-treated spleen cells were stained with fluorescent trinitrophenylated bovine serum albumin (FL-TNP-BSA). These cells were sorted, fluorescence negative from fluorescence positive, by using a multiparameter cell sorter and assayed for precursor frequency in antibody responses to TNP by limiting dilution analysis. The cells that bound FL-TNP-BSA were demonstrated to be enriched for antibody-forming precursors to the antigens TNP-lipopolysaccharide (LPS), TNP-sheep red blood cells (SRBC), and TNP- or DNP-Ficoll, whereas the fluorescence negative cells were depleted for these responses. B cells that bound FL-TNP-BSA were then sorted into populations that bound a moderate or high amount of FL-TNP-BSA. The B cells responsive to TNP-LPS and TNP-SRBC were present in both the moderate and high binding populations. In contrast, the B cells responsive to TNP- or DNP-Ficoll were present only in the cells that bound a moderate amount of FL-TNP-BSA. These experiments suggest that there is a population of B cells in adult mouse spleen that binds large amounts of antigen, and that can respond to antigen carried on LPS or SRBC but not carried on Ficoll.
D000818:Animals; D000941:Antigens; D001402:B-Lymphocytes / Q000145:classification*; D001665:Binding Sites; D001667:Binding, Competitive; D002417:Cattle; D002469:Cell Separation; D003208:Concanavalin A / Q000494:pharmacology; D004306:Dose-Response Relationship, Immunologic; D004912:Erythrocytes / Q000276:immunology; D005260:Female; D005455:Fluorescent Antibody Technique; D008070:Lipopolysaccharides / Q000276:immunology; D008264:Macrophages; D051379:Mice; D008805:Mice, Inbred A; D012710:Serum Albumin, Bovine; D012756:Sheep; D013194:Staining and Labeling; D014301:Trinitrobenzenes / Q000276:immunology
D000941:Antigens; D008070:Lipopolysaccharides; D014301:Trinitrobenzenes; D003208:Concanavalin A; D012710:Serum Albumin, Bovine