Purification of rabbit B spleen cells by removal of adherent and of T cells.
B lymphocytes from the rabbit spleen were freed of T cells by removal of cells which formed rosettes with papain-treated rabbit erythrocytes. Additional purification could be achieved if fractionation by rosette removal was preceded by removal with a magnet of cells which adhered to or ingested poly L-lysine coated iron core particles. Cell yield and purification were assessed by complement mediated cytotoxic kill of B and T cells with antibody directed against RABELA and RTLA, respectively. Other criteria depended on determination of the number of Fc receptor bearing cells and of thymidine uptake by cells which were stimulated with concanavalin A, PHA or with antibody directed against the allotypic specificity of receptor Ig light chains. Purified preparations of B cells were obtained in a yield of about 20% of the B cells in the original spleen and contained less than 10% of cells which were not B cells. This method allows purification which does not interfere with the membrane of the isolated cells.
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D002448:Cell Adhesion; D002469:Cell Separation; D003165:Complement System Proteins; D003208:Concanavalin A / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic; D005260:Female; D007141:Immunoglobulin Fc Fragments; D008297:Male; D010835:Phytohemagglutinins / Q000494:pharmacology; D011817:Rabbits; D012397:Rosette Formation; D013154:Spleen / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D013936:Thymidine / Q000378:metabolism
D007141:Immunoglobulin Fc Fragments; D010835:Phytohemagglutinins; D003208:Concanavalin A; D003165:Complement System Proteins; D013936:Thymidine