A simple method for specific depletion of B lymphocytes from bovine peripheral blood mononuclear cells.
The treatment of bovine peripheral blood mononuclear cells with rabbit anti-bovine immunoglobulin, goat anti-rabbit immunoglobulin (GAR) and complement resulted in the specific lysis of all surface immunoglobulin (SIg) bearing B lymphocytes. No SIg+ lymphocytes were detected after 12 hours of culture following lytic treatment, whereas cells treated with antibody without complement readily stained for SIg. The percentage of cells forming E-rosettes or binding peanut agglutinin (PNA) increased following lysis. The percentage of latex-ingesting monocytes also increased after lysis even though many of these cells had cytophilic Ig. B lymphocyte-depleted (T cell-enriched) populations cultured with phytohemagglutinin (PHA) and concanavalin A (Con A) were never less reactive than unseparated cells. No differences in background mitosis was observed for these 2 cells preparations. These results suggest that bovine SIg- cells do not require B lymphocytes to respond to PHA and Con A and that lysis of B lymphocytes does not alter the responsiveness of the recovered SIg- cells.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D001402:B-Lymphocytes; D002417:Cattle; D002469:Cell Separation; D003165:Complement System Proteins; D003208:Concanavalin A / Q000494:pharmacology; D006041:Goats; D008212:Lymphocyte Depletion; D010586:Phagocytes; D010835:Phytohemagglutinins / Q000494:pharmacology; D011817:Rabbits; D011947:Receptors, Antigen, B-Cell; D012397:Rosette Formation; D012756:Sheep
D000888:Antibodies, Anti-Idiotypic; D010835:Phytohemagglutinins; D011947:Receptors, Antigen, B-Cell; D003208:Concanavalin A; D003165:Complement System Proteins