Protective, anti-tumor monoclonal antibody recognizes a conformational epitope similar to melibiose at the surface of invasive murine melanoma cells.
Polyclonal and monoclonal antibodies (MAbs) have been raised against B16F10 cells collected from growing tumors in vivo or grown in culture media supplemented with normal mouse serum to avoid xenogeneic reactivity. Antibody binding to glutaraldehyde-fixed melanoma cells and Melan A melanocytes was assayed using chemiluminescent-enzyme-linked immunosorbent assay (CL-ELISA) for increased sensitivity. Most of the reactivity of antitumor polyclonal IgG (92%) was inhibitable by a carbohydrate pool consisting of melibiose, mannose, lactose, and sialic acid. Two monoclonal IgG(2a) antibodies, A4 and B11, had their reactivity to melanoma cells completely and specifically inhibited by melibiose. MAb A4 did not bind to alpha-galactosyl residues abundantly expressed in a protozoan mucin used as substrate, and its binding to the tumor cells was not affected by alpha-galactosidase treatment or addition of alpha-methyl-galactopyranoside or raffinose. Recognition of a mimotope similar to melibiose is suggested. MAb is cytotoxic in vitro in a complement-mediated reaction and effectively neutralizes melanoma cells protecting syngeneic mice against tumor development in vivo. This MAb is thus an important tool for further studies on antitumor adjuvant therapy combined with other agents associated with immuno- and chemotherapy of invasive melanoma.
Dobroff
A S
AS
Department of Microbiology, Immunology and Parasitology, Federal University of São Paulo (UNIFESP), São Paulo, Brazil.
Rodrigues
E G
EG
Moraes
J Z
JZ
Travassos
L R
LR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000912:Antibodies, Neoplasm / Q000008:administration & dosage; D000951:Antigens, Neoplasm; D019496:Cancer Vaccines / Q000737:chemistry; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000494:pharmacology; D002469:Cell Separation; D003165:Complement System Proteins; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005434:Flow Cytometry; D005690:Galactose / Q000737:chemistry; D005976:Glutaral / Q000494:pharmacology; D006801:Humans; D006825:Hybridomas; D007074:Immunoglobulin G / Q000378:metabolism; D007167:Immunotherapy; D007785:Lactose / Q000378:metabolism; D058965:MART-1 Antigen; D008358:Mannose / Q000378:metabolism; D008545:Melanoma / Q000378:metabolism*; D008546:Melanoma, Experimental; D008553:Melibiose / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D009077:Mucins / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D009363:Neoplasm Proteins / Q000494:pharmacology; D011487:Protein Conformation; D014349:Trypanosoma cruzi
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D002241:Carbohydrates; D000939:Epitopes; D007074:Immunoglobulin G; D058965:MART-1 Antigen; C548667:MLANA protein, human; C548668:Mlana protein, mouse; D009077:Mucins; D009363:Neoplasm Proteins; D003165:Complement System Proteins; D008553:Melibiose; D019158:N-Acetylneuraminic Acid; D007785:Lactose; D008358:Mannose; D005976:Glutaral; D005690:Galactose