MURINE IGG1 AND IGE MEMORY B-CELLS

Anti-dinitrophenyl IgG1 and IgE antibody production were examined in v itro. Splenic B cells from mice immunized in vivo at various intervals with dinitrophenylated proteins were cultured in vitro with splenic T cells and splenic antigen-presenting cells from mice immunized previo usly (2 weeks) with keyhole limpet hemocyanin. Anti-dinitrophenyl IgG1 and IgE antibodies were determined in the supernatants by ELISA. With out T cells and antigen-presenting cells or without antigen, no anti-d initrophenyl antibodies were detected. When the B cells were separated by panning with anti-murine IgG1 antibody, only the cells adhering to anti-IgG1 produced anti-dinitrophenyl IgG1 antibodies. Anti-dinitroph enyl IgE production was observed from cells adhering and from cells no t adhering to the anti-IgG1-coated plates, as expected. However, anti- dinitrophenyl IgE was also produced from cells which adhered to the an ti-IgG1-coated plates, but did not have surface IgE antibodies. Theref ore, cells without surface IgE adhering to anti-IgG1 were capable of p roducing anti-dinitrophenyl IgE antibodies. (C) 1994 Academic Press, I nc.