The influence of avidity on signaling murine B lymphocytes with monoclonal anti-IgM antibodies. Effects of B cell proliferation versus growth inhibition (tolerance) of an immature B cell lymphoma

We have investigated the effect of the avidity of a monoclonal anti-mouse IgM antibody (mAb) on its ability to activate normal B cells vs its capacity to inhibit the growth of an immature B lymphoma, BKS-2. A panel of seven mAb with specificities for four different domains of mu-heavy chain were selected and their relative avidities measured. Of the seven mAb studied, four antibodies (b7-6, Ak11, AK15, and DS1) were found to have relatively higher avidities than three others (AK17, 331.12, and Bet-2). Among them, the mAb b7-6 was highly efficient in inducing proliferation of normal B cells, whereas all others (including the remaining three high avidity mAb) were inefficient in inducing B cell proliferation. This failure cannot be simply related to the differences in their fine specificity because all the mAb became highly stimulatory after coupling to Sepharose beads. With one exception (DS1), avidity was a better predictor of the growth-stimulatory capacity when the antibodies were immobilized. Taken together, these findings suggested that although high affinity was important, there were other factors that might influence the mitogenic potential of a soluble anti-mu mAb. On the other hand, all anti-mu mAb irrespective of their avidity, fine specificity, and isotype were capable of effectively inhibiting the growth of BKS-2 cells. Furthermore, there was a direct correlation between the avidity and the dose of the mAb required to cause half-maximal inhibition of BKS-2 growth. The F(ab) fragments of high avidity mAb b7-6 failed to inhibit the growth of BKS-2 cells, which indicates that a minimal level of cross-linking of the membrane Ig receptor is necessary for causing growth inhibition in BKS-2 cells. Although there was no significant difference in the tolerogenic potential of soluble vs immobilized anti-mu mAb in directly inactivating BKS-2 cells, their growth-inhibitory capacity was remarkably different in the presence of rIL-5. Thus, rIL-5 was effective in partially overcoming the tolerogenic effect of soluble but not immobilized anti-mu mAb. Unlike IL-5, LPS rescued BKS-2 cells from the inhibitory effect of both soluble and immobilized anti-mu antibodies. Altogether, these results suggested that the ligand binding requisites for tolerance induction were less stringent than those for B cell activation. Furthermore, a high affinity interaction between membrane Ig receptor and its ligand appeared to generate a dominant negative signal that was not reversed by lymphokines.