A monoclonal antibody to a carbohydrate epitope expressed on glycolipid and on alpha3beta1 integrin on human esophageal carcinoma.
A mouse monoclonal antibody (MAb-9) produced by immunization with a human esophageal carcinoma cell line, TE-2 (derived from undifferentiated squamous cell carcinoma) reacted specifically with about 30% of esophageal carcinoma cell lines and tissue sections from clinical samples. MAb-9 showed minimal reactivity with normal esophageal tissue. (125)I, fluorescent or gold particle labeled MAb-9 bound to TE-2 cell surfaces. (125)I-radiolabeled MAb-9 was used to detect reactive material from cell extracts in Western blot. Treatment of TE-2 membrane proteins with neuraminidase, N-glycanase or O-glycanase reduced antigen detection. Treatment of cells with periodic acid destroyed antibody binding in ELISA. Lipid extracts from cell membranes, containing glycolipids, also reacted with MAb-9. MAb-9 was used to purify target antigen from detergent solubilized membrane proteins and the prominent bands from subsequent gel electrophoresis were trypsin digested and analyzed by mass spectrometry. Peptides from alpha(3) and beta(1) integrin chains were identified. These data indicate that alpha3beta1integrin is prominently expressed on certain esophageal carcinomas and that a specific carbohydrate unit is selectively displayed on the alpha(3) integrin subunit as well as on glycolipid on the cell surface. The alpha3beta1 integrin expressed on A-431 carcinoma cells does not display this carbohydrate epitope and is not detected by MAb-9. Thus, expression of the carbohydrate epitope is the basis for the tumor selective reaction of MAb-9 with a subset of esophageal carcinomas.
Jamasbi
Roudabeh J
RJ
Department of Public & Allied Health, 504 Life Sciences Building, Bowling Green State University, Bowling Green, OH 34303-0287, USA. rjamasb@bgnet.bgsu.edu
Stoner
Gary D
GD
Foote
Linda J
LJ
Lankford
Trish K
TK
Davern
Sandra
S
Kennel
Stephen J
SJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D015153:Blotting, Western; D002241:Carbohydrates / Q000737:chemistry / Q000276:immunology*; D002277:Carcinoma / Q000378:metabolism*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002855:Chromatography, Thin Layer; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D004938:Esophageal Neoplasms / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D006017:Glycolipids / Q000737:chemistry* / Q000276:immunology; D006596:Hexosaminidases / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D039201:Integrin alpha3beta1 / Q000737:chemistry*; D008055:Lipids / Q000737:chemistry; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D009439:Neuraminidase / Q000737:chemistry / Q000378:metabolism; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D010504:Periodic Acid / Q000737:chemistry; D011817:Rabbits
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D000939:Epitopes; D006017:Glycolipids; D039201:Integrin alpha3beta1; D008055:Lipids; D010455:Peptides; D010504:Periodic Acid; D006596:Hexosaminidases; D009439:Neuraminidase; C060638:glycopeptide alpha-N-acetylgalactosaminidase; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase