Diversity in immunoreactivity of tumor-derived cytokeratin monoclonal antibodies.
The cytokeratins 8, 18, and 19, expressed in many normal and malignant epithelial cells, were purified from human gastrointestinal tumors and used as immunogen for hybridoma generation. The reactivity pattern of five of the generated ten monoclonal antibodies (MAb) was characterized biochemically and immunohistochemically. All of the generated MAb were reactive with the central rod portion of the cytokeratins, as determined after partial enzymatic degradation, and displayed characteristic reactivity patterns. MAb TS 4 exhibits pan-epithelial immunohistochemical reactivity staining of all epithelial structures, including all layers of epidermis and non-keratinizing squamous epithelium and myoepithelial cells. The determinant involved is present on several different cytokeratins, i.e., nos. 1, 5, 7, 8, and 15, as determined by immunoblotting experiments from different tissues and cell lines. MAb TS 1, TS 3, and TS 7 reveal pluri-epithelial reactivity pattern immunohistochemically, similar to TS 4, but they are unreactive with whole epidermis and with superficial cell layers of non-keratinizing squamous cells. MAb TS 1 was found to be highly specific and reactive only with cytokeratin 8. Furthermore, the TS 1 MAb alone can precipitate the antigen, indicating reactivity with repetitive epitopes on cytokeratin 8. MAb TS 3 and 7 bind to cytokeratins 7 and 8. Finally, MAb TS 8 was found to be immunohistologically the most restricted, in general lacking reactivity to hepatocytes, pancreatic and salivary gland acinar cells, proximal renal tubules, and luminal cells of the epididymis. TS 8 was mainly reactive with cytokeratin 19 and showed weak binding to cytokeratin 8 and 14.
Sundström
B E
BE
Department of Physiological Chemistry, University of Umeå, Sweden.
Nathrath
W B
WB
Stigbrand
T I
TI
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001667:Binding, Competitive; D015153:Blotting, Western; D002277:Carcinoma / Q000648:ultrastructure*; D002460:Cell Line; D003599:Cytoskeleton / Q000276:immunology*; D004848:Epithelium / Q000648:ultrastructure; D000939:Epitopes; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D007150:Immunohistochemistry; D007382:Intermediate Filaments / Q000276:immunology*; D007633:Keratins / Q000276:immunology*; D010232:Paraffin; D010920:Placenta / Q000648:ultrastructure; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005404:Fixatives; D005557:Formaldehyde; D007633:Keratins; D010232:Paraffin