title: Triggering of human peripheral blood B cells: polyclonal induction and modulation of an in vitro PFC response.
abstract: In the present studies we utilized polyclonal activators to analyze the signals required to induce antibody synthesis by subpopulations of human peripheral blood lymphocytes. The in vitro synthesis of antibody directed against sheep erythrocytes was measured by a modification of the Jerne plaque assay. The plaques observed contained central lymphoid cells, could be inhibited by anti-human immunoglobulin, and were strictly dependent on both the addition of complement and viable B lymphocytes actively synthesizing protein. Kinetic analysis showed that the PFC response developed over time during cell culture with peak activity occurring between days 6 and 7. It was found that human peripheral blood B lymphocytes, isolated by Sephadex G-200 anti-Fab column chromatography, were nonspecifically induced by either PWM or soluble products of antigen-activated human T cells to differentiate into PFC. In contrast, unfractionated peripheral blood lymphocytes. Ig- cells isolated from immunoabsorbant columns, and B cells enriched by nylon wool adherence were not triggered to antibody synthesis even in the presence of polyclonal activators. B cells isolated by nylon wool adherence and further fractionated by binding and elution from anti-Fab columns were then readily triggered by PWM to differentiate into PFC. These results suggest that the interaction of human B cells with anti-Fab columns, presumably at the Ig receptor, serves as a signal important in the differentiation events leading to PFC activity. The anti-Fab interaction alone, although critical, was not sufficient to induce the differentiation of precursor cells into PFC since subsequent triggering by antigen-activated T cell supernatants or PWM was required for the development of an optimal PFC response. Additional experimental data demonstrated that the PFC response could be regulated by autologous peripheral Ig- lymphoid cells in vitro. At low concentrations of PWM, Ig- cells consistently augmented the B cell PFC response whereas at high concentrations of PWM, suppression was observed. Depletion of E rosetting cells from the Ig- population eliminated the subset responsible for augmentation but did not eliminate the suppressor cell activity.
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D002474:Cell-Free System; D002850:Chromatography, Gel; D006462:Hemolytic Plaque Technique; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D007700:Kinetics; D037102:Lectins / Q000494:pharmacology; D012756:Sheep; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D007140:Immunoglobulin Fab Fragments; D037102:Lectins