IL-4-dependent IgE class switching in an anti-trinitrophenyl B-cell hybridoma after engagement of antigen receptors

Abstract

A B-cell hybridoma, TP67.21 that expresses surface anti-trinitrophenyl (TNP) IgM but does not secrete the antibody spontaneously has been reported to differentiate into anti-TNP IgM-secreting cells in response to lipopolysaccharide or engagement of surface IgM. Here, we report isolation and characterization of a subclone, TP67.21E (TP.E) that undergoes isotype switching to IgE in an interleukin (IL)-4-dependent manner. TP.E cells secreted anti-TNP IgE depending on exogenous IL-4 when they were cultured with an anti-IgM antibody for 6–8 days. 8-Mercaptoguanosine, which has been shown to enhance IgE class switching in murine splenic B-cells further augmented the IgE response in TP.E cells. Immunofluorescence microscopy revealed that approximately 1.2% of the cultured cells became positive for intracellular IgE after the stimulation culture. The germline ε transcripts were expressed transiently on days 2–4 of the culture, while expression of the productive ε transcripts was induced 5 days after the start of the culture, thus suggesting that IgE class switching occurred in TP.E cells under these conditions.