title: The interaction of specific T-cell help and non-specific B-cell growth factors in the production of anti-tetanus antibody by human B cells grown in serum-free microcultures.
abstract: Apart from a brief period after in-vivo immunization, only a minority of human donors provide peripheral lymphocytes that synthesize specific antibody on stimulation with tetanus toxoid in vitro. A 20 microliters hanging drop microculture technique using serum-free medium has been adapted to analyse the conditions under which B cells mature into antibody-secreting cells. Multiple permutations of antigen dose, cell concentration and T:B cell ratios have been examined. The results indicate that in-vitro failure of antigen response by the majority of donors is not due simply to an inappropriate choice of culture conditions. The addition to antigen-stimulated cultures of a lectin-free conditioned medium derived from pokeweed mitogen-stimulated peripheral lymphocytes, enables B cells from the majority of donors to produce high titres of specific antibody, in a T-dependent manner, for up to 24 months after immunization. The observed failure of prolonged antigen responsiveness in vitro thus appears to represent a failure to expand a population of antigen-specific B cells, rather than indicating an absence of such clones.
mesh_terms: D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D001402:B-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D003470:Culture Media; D006133:Growth Substances / Q000276:immunology*; D006801:Humans; D007376:Interleukin-2 / Q000276:immunology*; D015847:Interleukin-4; D013601:T-Lymphocytes / Q000276:immunology; D013745:Tetanus Toxoid / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D003470:Culture Media; D006133:Growth Substances; D007376:Interleukin-2; D013745:Tetanus Toxoid; D015847:Interleukin-4