Characterization of a pan-species reactive monoclonal antibody specific for a cell surface epitope which could serve as a marker for human monocytic and megakaryocytic differentiation.
A mouse IgG(1)-producing hybridoma, CSC-31, was isolated and characterized. The monoclonal antibody (MAb) was originally raised against monkey kidney cell-surface molecules. FACS analysis further showed that CSC-31 exhibited broad tissue and species reactivity. Although most human T- and B-cell lines failed to react with CSC-31, myeloid, and erythroleukemia cells lines such as THP-1 and K562 expressed the CSC-31 cell surface marker. Furthermore, in vitro differentiation of HL-60, U-937, and K562 showed that expression of the CSC-31 marker is associated with monocytic or megakaryocytic differentiation. However, up-regulation of the CSC-31 marker expression was not detected during granulocytic or erythroid differentiation. Through the in vitro differentiation of K562, it was demonstrated that up-regulation of the CSC-31 marker required novel PKCs and might be regulated by the MAPK signaling pathway. Last, limited biochemical analysis demonstrated that the CSC-31-specific epitope is sensitive to digestion by papain yet highly resistant to other proteases.
Chang
Chew Shun
CS
John P. Robarts Research Institute, The University of Western Ontario, London, Ontario, N6G 2V4, Canada.
McFadden
Grant
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000954:Antigens, Surface / Q000276:immunology; D015415:Biomarkers; D002454:Cell Differentiation / Q000276:immunology* / Q000502:physiology; D017077:Culture Media, Conditioned / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D006801:Humans; D008533:Megakaryocytes / Q000276:immunology* / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009000:Monocytes / Q000276:immunology* / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism; D015854:Up-Regulation
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D015415:Biomarkers; D017077:Culture Media, Conditioned; D000939:Epitopes; D011493:Protein Kinase C; D020928:Mitogen-Activated Protein Kinases; D013755:Tetradecanoylphorbol Acetate