Anti-immunoglobulin-induced proliferative response of rabbit lymphocytes in a serum-free medium.
The aim of this study was to determine if rabbit lymphocytes were activated with a specific ligand, anti-immunoglobulin (anti-Ig) in a well-defined serum-free medium. A modified medium, composed of Neuman-Tytell (NT) basal medium supplemented with BSA, transferrin (Tf), and fatty acids (FA), was found to support a vigorous anti-Ig-induced proliferative response of rabbit lymphocytes. The optimal concentrations of BSA, Tf and FA supplements were determined for the mitogen-induced response. The mitogenesis was enhanced by addition of 2-mercaptoethanol. The proliferation could be maintained for 10 days provided cells received fresh medium and mitogen on day 5. The presence of anti-Ig throughout the culture period is required for this extended proliferation. The response did not require the Fc portion of the anti-Ig and was blocked by soluble rabbit F(ab')2 fragments. The anti-Ig-activated blasts lacked a T cell surface marker and about half of them re-expressed sIg when anti-Ig was washed out (assayed 24 h later). The viability of the mitogen-induced cells on day 5 was above 55%, whereas the viability of the cells without mitogen was 36%. The supplemented NT medium could also support Con A-induced mitogenesis of rabbit lymphocytes. Additionally, Con A- and LPS-stimulated murine lymphocytes showed considerable proliferation for up to 5 days in the supplemented NT medium.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000502:physiology*; D001790:Blood Physiological Phenomena; D002417:Cattle; D003470:Culture Media; D004306:Dose-Response Relationship, Immunologic; D005227:Fatty Acids / Q000494:pharmacology; D005260:Female; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007700:Kinetics; D008213:Lymphocyte Activation; D008297:Male; D008623:Mercaptoethanol / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013154:Spleen / Q000166:cytology; D014168:Transferrin / Q000494:pharmacology
D000888:Antibodies, Anti-Idiotypic; D003470:Culture Media; D005227:Fatty Acids; D007140:Immunoglobulin Fab Fragments; D014168:Transferrin; D012710:Serum Albumin, Bovine; D008623:Mercaptoethanol