Mouse peritoneal macrophages cultured serum-free deposit complement on IgM-coated sheep erythrocytes in vitro

We conclude that the macrophages during cultivation produce the complement components of the classical pathway of complement, deposit complement components on EIgM and then phagocytose these cells via complement receptors. The conclusion is based on the following: EIgM, an activator of the classical pathway, are ingested when cultured serum-free with mouse peritoneal macrophages. We found a significantly higher binding of labelled protein to EIgM than to E kept in macrophage cultures in the presence of tritiated leucine, showing that de novo synthesis of macrophage-derived protein with affinity to EIgM takes place. A fraction of the bound protein is C3b and iC3b, since anti-mouse C3 antibodies bound to the co-cultured EIgM. Cycloheximide or anti-Mac-1 in the cultures inhibited macrophage attachment and uptake of EIgM. The phagocyte uptake of EIgM coated with complement by serum pretreatment was not inhibited by cycloheximide. This shows that the phagocytosis of the EIgM is dependent on erythrocyte-bound complement proteins made by the macrophage.