Immunohistological characterization of mucin epitopes by pre-treatment of gastro-intestinal sections with periodic acid.
Periodate pretreatment of paraffin sections of ethanol-fixed gastrointestinal mucosae was used to characterize the carbohydrate or peptidic nature of mucin epitopes by immunoperoxidase. Immunoreactivity of monoclonal antibodies (MAbs) against histo-blood group related carbohydrate epitopes such as A, Lea, Lec, Sialosyl Lea, H type 2, I, T, Tn and sialosyl Tn dramatically decreased or even disappeared after periodate pretreatment of deparaffinized sections. In contrast, the immunoreactivity of MAbs against peptide mucin epitopes such as the gastric M1 mucin epitopes was almost unaffected by this treatment. Moreover, periodate treatment revealed cryptic peptide M1 mucin epitopes and the peptide MUSE11 epitope associated with the 20 amino acid tandem repeat (PDTRPAPGSTAPPAHGVTSA). An increase of cross-reactions of anti-human M1 MAbs with gastric epithelium of different vertebrate species was detected with periodate treatment. Our results suggest that this method can be useful for preliminary characterization of the biochemical nature of mucin epitopes (peptidic or saccharidic) and for demasking peptidic tumour markers which are hidden by saccharide molecules in normal tissues.
Bara
J
J
Mucin Immunochemistry Laboratory, SDI-6316, Villejuif, France.
Decaens
C
C
Loridon-Rosa
B
B
Oriol
R
R
D000293:Adolescent; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002240:Carbohydrate Sequence; D003429:Cross Reactions; D000939:Epitopes / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D005753:Gastric Mucosa / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000378:metabolism*; D008875:Middle Aged; D008969:Molecular Sequence Data; D009077:Mucins / Q000096:biosynthesis* / Q000276:immunology; D010504:Periodic Acid; D014714:Vertebrates
D000911:Antibodies, Monoclonal; D000939:Epitopes; D009077:Mucins; D010504:Periodic Acid