IgM and IgG secretion in human B-cell lines regulated by B-cell-inducing factors (BIF) and phorbol ester

Human B-cell lines were screened for stimulation of immunoglobulin production by incubation with lymphokine (LK) or tumor promoter, phorbol myristic acetate (PMA). One group of lines had essentially no immunoglobulin-secreting cells (ISC) under any condition (less than 0.01%), detected by a reverse plaque assay. Another group of lines had high levels of ISC (>5%) which was not increased substantially by inducing agents. In a third group of IgM and IgG lines, there were intermediate levels of ISC which could be increased by LK, PMA or both agents. No evidence for isotype switching in a number of stimulated IgM and IgG cell lines was detected. Clone SKW6.4 of an IgM line was highly responsive to a B-cell-inducing factor (BIF) in LK. BIF for SKW6.4 and IgG line ARH-77 was weakly binding to DEAE cellulose, about 20,000 mol. wt., and separable from IL-2 by blue agarose chromatography. IL-2 did not stimulate secretion in SKW6.4 with or without field BIF. In Clone SKW6.4, BIF stimulated ISC per recovered cell up to 30-fold by day 1 of culture, and these plateau levels of about 6% ISC were maintained for longer than 4 days. Treatment of cells with BIF for less than 1 day was sufficient to produce maximum effect on this clone for the succeeding 4 days. Cells stimulated with BIF and then subcultured at day 3 without BIF showed ISC numbers increasing but at a slower rate than the total population, suggesting that the induced differentiation state is long-lived (half-life of % ISC >6 days) and that ISC produce some daughter ISC. In declining cultures readdition of BIF boosted ISC levels again to about 6%. The continual presence of 20K BIF for 12 days had no apparent effect on total cell growth. In conclusion, a number of cell lines are sensitive to stimulation of immunoglobulin secretion and may provide models for induction of human antibody production.