Rat monoclonal antibodies produced against rat colorectal adenocarcinomas define tumor- and colon-associated, auto-immunogenic antigens.
Monoclonal antibodies (MAbs) were derived from rats immunized against allo- and syngeneic rat colon carcinomas. Screening was performed by immunohistochemistry modified for MAbs of the same species as the tissue used for frozen sections. From 2 fusions, several MAbs were found that bound to syngeneic tumor tissue but showed little or no staining of normal adult tissues. Another group of MAbs demonstrated an intracellular staining of tumor cells and staining of mucus and goblet cells in the distal 2/3 of the normal colon. A third group of MAbs showed staining of both tumor and normal colon tissue. Staining patterns were reproduced with 6 MAbs selected from the first 2 groups after purification and biotinylation. One MAb, 10B12, recognized an antigen expressed in 10/11 colon carcinomas, the lowest parts of colonic crypts, intracellularly in a subpopulation of pancreatic acinar cells and mucus in antrum. It was used for affinity purification of a high-molecular-weight antigen, to which 3 of the other rat MAbs also bound. This antigen was also bound by antibodies to blood group A and isogloboseries carbohydrate determinants. Competition with the labelled 10B12 MAb for binding to the purified antigen was demonstrated in sera of tumor-bearing and immune rats. Thus, this high-molecular-weight glycoprotein expressed both auto-antigenic, tumor-associated and tissue-type-restricted epitopes. The 10B12 MAb homogeneously stained the majority of colorectal carcinomas and the antigenic determinant was expressed on the cell surface of 12/13 tissue-cultured colon-carcinoma clones. Modulation of the cell-surface antigen phenotype by recombinant rat interferon-gamma markedly increased expression of this determinant.
Department of Tumor Immunology, University of Lund, Sweden.
N T
NT
Brodin
B
B
Jansson
H O
HO
Sjögren
D019813:1,2-Dimethylhydrazine; D000230:Adenocarcinoma / Q000139:chemically induced / Q000276:immunology* / Q000473:pathology; D000818:Animals; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000032:analysis*; D000954:Antigens, Surface / Q000032:analysis; D001324:Autoantigens / Q000032:analysis*; D002273:Carcinogens; D002460:Cell Line; D015179:Colorectal Neoplasms / Q000139:chemically induced / Q000276:immunology* / Q000473:pathology; D004127:Dimethylhydrazines; D000939:Epitopes / Q000032:analysis; D005260:Female; D015151:Immunoblotting; D007371:Interferon-gamma / Q000494:pharmacology; D008297:Male; D011863:Radioimmunoassay; D051381:Rats; D011914:Rats, Inbred BN; D011920:Rats, Inbred WF
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D001324:Autoantigens; D002273:Carcinogens; D004127:Dimethylhydrazines; D000939:Epitopes; D007371:Interferon-gamma; D019813:1,2-Dimethylhydrazine