The effects of bacterial lipopolysaccharide, anti-receptor antibodies and recombinant interferon on mouse B cell cycle progression using 5-bromo-2'-deoxyuridine/hoechst 33258 dye flow cytometry

Polyclonal stimulation of resting B cells with anti-antigen receptor antibodies [anti-IgM mu chain antibody (anti-mu)] or with bacterial lipopolysaccharide (LPS) stimulates a proportion of B cells to proliferate. Exposure of resting B cells to both LPS and anti-mu activates a larger population of resting B cells than either alone, suggesting a synergistic effect of these two stimuli. Although recombinant interferon (rIFN) either alone or in combination with anti-mu has no apparent proliferative activity (as measured by [3H]thymidine incorporation), application of 5-bromo-2'-deoxyuridine/Hoechst 33258 dye flow cytometry reveals a distinct effect of rIFN on B cell growth. In the presence of anti-mu plus LPS, rIFN causes the majority of B cells to enter the cell cycle (CC), but a subset of B cells remains in the resting stage. Another subset of B cells has extremely rapid CC transit times, with a CC duration of less than 10 h. These studies show that both anti-mu and LPS are competence factors (which move cells from the G0 phase to the G1 phase). LPS acts also as a CC progression factor, while rIFN is a CC potentiating factor.