Induced Autophagocytosis in Macrophages. Origin of the Segregating Membranes

Cultured mouse peritoneal macrophages were exposed for 48 h to a large and toxic dose of polyacrylamide microspheres (previously designed for use as a lysosomotropic carrier for the intracellular delivery of enzymes and other macromolecules). This treatment induced autophagocytosis in the macrophages, which contained abundant autophagic vacuoles at 24 h post exposure. Transmission electron microscopical studies including enzyme cytochemistry showed that the segregating membranes in autophagosome formation consisted of flattened, smooth-surfaced vacuoles with a granular matrix in which reaction product indicating acid phosphatase activity could be demonstrated. The autophagic vacuole formation was apparently effected by wrapping of a portion of the cytoplasm in a sheet formed by flattening and fusion of multiple small vacuoles with acid phosphatase activity in their matrices. The conclusion is drawn that the segregating membranes are derived from lysosomes or GERL structures in this particular system of induced autophagocytosis.