The mechanisms by which human oligodendroglial cells, KG-1-C cells, phagocytose mycobacteria, especially Mycobacterium leprae, were studied. The ability of glial cells to phagocytose M. leprae was inhibited by azide, dinitrophenol (inhibitors of oxidative phosphorylation), and iodoacetamide but not fluoride (both are inhibitors of glycolysis). Thus, the energy metabolism dependency is somewhat different from that of peritoneal macrophages and polymorphonuclear leukocytes, the phagocytic capacities of which are mainly dependent on glycolysis. Phagocytosis of M. leprae by KG-1-C cells was markedly suppressed by a microfilament inhibitor (cytochalasin B) but not microtubule inhibitors (colchicine and vinblastine), as with macrophages. The phagocytosis of M. leprae by KG-1-C cells was dependent on the lipid and somewhat on the sugar ligands of the organism. Moreover, the phagocytosis of a given mycobacterium by KG-1-C cells correlated well with its hydrophobicity, thus revealing the importance of some lipid moieties on the surface of bacteria in the establishment of rigid binding interaction of bacteria with KG-1-C cells, before the onset of engulfment. Electric charge of a given microorganism did not correlate with its phagocytosis by KG-1-C cells.