title: Frequency analysis of human peripheral blood B cells producing IgM-rheumatoid factor. Differential effects of stimulation with monoclonal antibodies to CD3 and Staphylococcus aureus.
abstract: The extent and nature of IgM-rheumatoid factor (RF) precursors within normal human B cells were examined by utilizing two different polyclonal B cell stimulators, Staphylococcus aureus Cowan I (SA) and immobilized mAb to the CD3 molecular complex (64.1). In cultures stimulated with SA, B cells produced IgM-RF in the presence of T4 cells, factors generated from mitogen-activated T cells (TF), or IL-2. Similarly, in cultures stimulated with immobilized anti-CD3, T4 cells that had been treated with mitomycin C (T4 mito) induced the production of large amounts of IgM-RF. Limiting dilution analyses revealed that the precursor frequencies of IgM-RF-producing cells induced by SA + TF and by immobilized anti-CD3-activated T4 mito were 0.008 +/- 0.001/100 B cells (n = 7) and 0.043 +/- 0.004/100 B cells (n = 6) (mean +/- SEM), respectively. Of note, the proportion of IgM-secreting cells that produced IgM-RF was much greater in cultures stimulated with SA + TF (30 to 61%) than that noted in cultures containing immobilized anti-CD3-stimulated T4 mito (1.0 to 3.9%). When B cells were co-stimulated with both SA and immobilized anti-CD3-activated T4 mito, the frequency of IgM-RF producing cells increased further to 0.12 to 0.27/100 B cells (4.6 to 21.2% of IgM-producing cells). These results indicate that both SA and immobilized anti-CD3 are potent stimulators of IgM-RF precursors. Moreover, the combination of SA and immobilized anti-CD3 provides a very potent in vitro signal for IgM-RF elaboration, inducing the production of this autoantibody from 1 to 3 in 1000 circulating normal B cells.
authors: Department of Medicine and Physical Therapy, University of Tokyo School of Medicine, Japan.
authors: S
authors: S
authors: Hirohata
authors: T
authors: T
authors: Inoue
authors: T
authors: T
authors: Miyamoto
mesh_terms: D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte / Q000502:physiology; D001402:B-Lymphocytes / Q000276:immunology*; D017252:CD3 Complex; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D066298:In Vitro Techniques; D007958:Leukocyte Count; D008213:Lymphocyte Activation; D011043:Pokeweed Mitogens; D011948:Receptors, Antigen, T-Cell / Q000502:physiology; D012217:Rheumatoid Factor / Q000096:biosynthesis*; D013211:Staphylococcus aureus / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011043:Pokeweed Mitogens; D011948:Receptors, Antigen, T-Cell; D012217:Rheumatoid Factor