Human monocyte-mediated lysis of antibody-coated tumor cells: the role of the cytoskeleton in monocytes.
Human monocytes (M phi) show high cytolytic activity towards antibody-coated tumor cells (AbK562). In this report, the relationship between the cytoskeleton in the M phi and the M phi cytolytic activity has been investigated. The actin filament inhibitors cytochalasin B and dihydrocytochalasin B (H2CB) both reduced M phi-mediated lysis of AbK562 cells by approximately 50% at a concentration of 1 microM. This concentration of H2CB did not inhibit the number of target cells bound to M phi. Dihydrocytochalasin B did not inhibit the M phi ability to release cytotoxic protein factors, suggesting that H2CB does not inhibit lysis by inhibiting release of cytotoxic protein factors. Immunofluorescence microscopy showed a rapid accumulation of actin filaments towards the contact area in more than 80% of the examined M phi-AbK562 conjugates. Exposure to H2CB did not prevent this accumulation, but caused aggregation of the accumulated actin filaments in the contact area with the target cell. Accumulation of actin filaments did not occur toward tumor cells not coated with antibodies. Scanning and thin section electron microscopy demonstrated large M phi pseudopodia directed toward the AbK562 cells, with close apposition of the effector and target cell membranes with interdigitations. The formation of the M phi pseudopodia was inhibited by exposure to H2CB. These observations indicate that M phi membrane motility toward AbK562 cells is closely related to M phi-mediated lysis of AbK562 cells. Immunofluorescence microscopy of the microtubule-organizing center (MTOC) and the Golgi apparatus revealed that both the MTOC and the Golgi apparatus in M phi reoriented towards the bound AbK562 cells in approximately 45% of the examined M phi-AbK562 conjugates. The microtubule-depolymerizing drugs colchicine and vinblastine did not inhibit M phi-mediated lysis of AbK562 cells at concentrations which disrupted the microtubule arrays in the M phi. The carboxylic ionophore monensin, which blocks Golgi-derived secretion, inhibited M phi-mediated lysis of AbK562 to a lesser extent as compared to H2CB. These results suggest that microtubule functions are of less importance in M phi-mediated lysis of AbK562 cells as compared to actin filament functions. However, the MTOC and the Golgi apparatus could participate in M phi-mediated lysis of AbK562 cells by mechanisms related to secretion of cytotoxic molecules.
D000199:Actins / Q000032:analysis / Q000502:physiology; D000328:Adult; D000818:Animals; D000912:Antibodies, Neoplasm / Q000502:physiology; D000920:Antibody-Dependent Cell Cytotoxicity / Q000187:drug effects; D003599:Cytoskeleton / Q000032:analysis / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000502:physiology; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000276:immunology* / Q000648:ultrastructure; D008870:Microtubules / Q000187:drug effects / Q000502:physiology; D008985:Monensin / Q000494:pharmacology; D009000:Monocytes / Q000276:immunology* / Q000502:physiology / Q000648:ultrastructure; D011817:Rabbits
D000199:Actins; D000912:Antibodies, Neoplasm; D008985:Monensin