title: The involvement of the major surface glycoprotein (gp63) of Leishmania promastigotes in attachment to macrophages.
abstract: The interaction between the macrophage and the parasite plays a central role in the continued success of Leishmania infection. The promastigote surface ligand, and its complementary macrophage membrane receptor, involved in attachment and phagocytosis are likely to exert considerable influence over the outcome of a new infection. In this study, we report experiments pertaining to one such parasite membrane protein. Initial examination of promastigote surface proteins by radiolabeling and two-dimensional-polyacrylamide gel electrophoresis revealed an abundant polypeptide with an apparent m.w. of 63,000. Lectin-binding studies indicated that it was a glycoprotein containing mannose, N-acetyl glucosamine, and N-acetyl galactosamine residues. Monospecific antiserum raised against this glycoprotein, gp63, decorated the entire promastigote plasmalemma. Univalent antibody fragments from this antiserum blocked the interaction between promastigotes and macrophages by inhibiting attachment. Anti-gp63-inhibition reduced parasite/macrophage binding to 30 to 35% of the control binding level. Additional evidence of the involvement of gp63 in attachment to macrophages was provided by studies that made use of gp63-containing proteoliposomes. These vesicles were avidly phagocytosed by macrophages. Uptake of the gp63-containing liposomes was suppressed by greater than 90% by both anti-gp63 F(ab) fragments and the oligosaccharide mannan, indicating that their phagocytosis was receptor dependent. These results demonstrate that the abundant glycoprotein gp63 plays an important role in attachment of promastigotes to macrophages, and attachment via this parasite ligand is sufficient to trigger phagocytosis.
mesh_terms: D000268:Adhesiveness; D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology* / Q000302:isolation & purification; D000954:Antigens, Surface / Q000276:immunology* / Q000302:isolation & purification; D001667:Binding, Competitive; D006023:Glycoproteins / Q000276:immunology* / Q000302:isolation & purification; D007106:Immune Sera / Q000032:analysis; D007140:Immunoglobulin Fab Fragments / Q000502:physiology; D007894:Leishmania mexicana / Q000254:growth & development / Q000276:immunology*; D008081:Liposomes / Q000276:immunology; D008264:Macrophages / Q000276:immunology* / Q000469:parasitology; D051379:Mice; D008807:Mice, Inbred BALB C; D008808:Mice, Inbred CBA; D010587:Phagocytosis; D011510:Proteolipids / Q000276:immunology
chemical_list: D000953:Antigens, Protozoan; D000954:Antigens, Surface; D006023:Glycoproteins; D007106:Immune Sera; D007140:Immunoglobulin Fab Fragments; D008081:Liposomes; D011510:Proteolipids