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.