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  • 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.
    Author: Hirohata S, Inoue T, Miyamoto T.
    Journal: J Immunol; 1990 Sep 15; 145(6):1681-6. PubMed ID: 2144005.
    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.
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