Binding and processing of immunostimulatory Fc gamma 1 fragments by the murine macrophage cell line P388D1.

Previous data from this laboratory indicated that human Fc gamma fragments induce murine B cells to proliferate and that the induction is macrophage-dependent. To further investigate the role of macrophages in this phenomenon, biologically active Fc gamma fragments from a human IgG1 myeloma protein and the murine macrophage-like cell line P388D1 were utilized. Fc gamma 1 fragments bound specifically and to a single class of receptor on P388D1 cells with a Ka value of 4 X 10(6) M-1 and to approximately 2.4 X 10(5) binding sites/cell. The binding was not effectively inhibited by two immunostimulatory Fc gamma 1 subfragments that were macrophage independent, i.e., pFc' fragments approximating the C gamma 3 domain of IgG1 and synthetic peptides representing residues 335-357 in IgG1. P388D1 cells were able to process Fc gamma 1 fragments but not intact IgG1 into subfragments that were able to induce lymphocyte proliferation in the absence of macrophages. The processing was rapid and resulted in active subfragments of several size classes. These findings not only further document the molecular and cellular events in these systems but underscore the usefulness of the P388D1 cell line in future studies on Fc fragment-induced lymphocyte regulation.