Abstract To appreciate better the mechanisms by which B lymphocytes are activated by anti-Ig antibodies, we characterized seven monoclonal mouse allo-antibodies to IgD of the a allotype for their isotypes, fine specificities, IgD-cross-linking abilities, avidities, and abilities to activate B cells in vitro and in vivo. Three of the monoclonal antibodies tested bound to the Fc fragment of IgD with relatively high avidity and were effective at cross-linking IgD, since they precipitated soluble IgD and rapidly capped B cell membrane IgD. These were the only antibodies tested that induced B cell DNA synthesis in vitro and were the most effective antibodies at inducing in vivo increases in B cell size and DNA synthesis and in vitro and in vivo increases in B cell surface Ia expression. Two antibodies bound to the Fd fragment of IgD with relatively high avidity but could not rapidly cap cell membrane IgD or precipitate soluble IgD even in the presence of 2% polyethylene glycol. These high-avidity, poorly cross-linking antibodies were unable to stimulate B cell DNA synthesis in vitro and were much less effective than the first group of anti-delta antibodies at stimulating in vivo increases in B cell DNA synthesis, size, or surface Ia expression or in vitro increases in surface Ia expression. One antibody, which bound to the Fc fragment of IgD with an intermediate avidity, was unable to rapidly cap B cell membrane IgD or precipitate soluble IgD in saline, but could precipitate soluble IgD in the presence of 2% polyethylene glycol. This antibody failed to induce B cell DNA synthesis in vitro and was as effective as the higher-avidity, poorly cross-linking antibodies at stimulating increases in B cell size, surface Ia expression, and DNA synthesis in vivo, and surface Ia expression in vitro. One antibody, which bound to the Fd fragment of IgD with low avidity and was unable to precipitate soluble IgD or to cap cell membrane IgD, had little ability to activate B cells by any of the parameters studied. Each of the monoclonal anti-delta antibodies, regardless of isotype or fine specificity, when bound to agarose to increase its ability to cross-link IgD, was mitogenic for B cells in vitro. None of the monoclonal antibodies to IgD of the a allotype stimulated B cells from b allotype mice to increase their size, surface Ia expression, or synthesis of DNA in vitro or in vivo.Our studies suggest that 1) the ability to cross-link surface Ig is the primary determinant of an anti-Ig antibody’s ability to activate B cells; 2) avidity and fine specificity influence the B cell-activating ability of an anti-IgD antibody by affecting its ability to effectively crosslink cell membrane IgD: 3) the induction of B cell DNA synthesis inv itro requires more stringent swface Ig cross-linking than does the induction of increased B cell surface Ia expression; and 4) even ligands that are ineffective at cross-linking B cell surface Ig and are non-mitogenic in vitro are able to stimulate some B cells to synthesize DNA in vivo.

Goroff, D. K., Stall, A., Mond, J. J., & Finkelman, F. D. (1986). In vitro and in vivo B lymphocyte-activating properties of monoclonal anti-delta antibodies. I. Determinants of B lymphocyte-activating properties. The Journal of Immunology, 136(7), 2382–2392.