Identification and characterization of a mouse cell surface antigen with alternative molecular forms

Abstract

We present the characterization of a new mouse cell surface protein, recognized by the 3E8-specific monoclonal antibody. The expression of this antigen is predominantly restricted to the hematopoietic and lymphoid tissues: bone marrow, spleen, lymph node, and thymus. Immunoblot analyses show that the 3E8 determinant is present on molecules with different apparent relative masses. The 3E8 antigen migrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a single band of Mr 115,000 for normal nonstimulated spleen cells and thymocytes and as two bands of Mr 115,000 and Mr 125,000 for bone marrow cells and mitogen-stimulated spleen cells. The multiple sizes of the 3E8 antigens (isoforms) found on various cell lines are not due to allelic polymorphism, but instead may reflect the specific cell type or reflect the cell's state of activation or maturation. Results from lectin chromatography and N-glycanase and neuraminidase studies suggest that the 3E8 antigen is a heavily sialylated O-linked glycoprotein. The unusual features of this antigen indicate that it may be the mouse homologue of the rat W3/13 antigen and the human leukosialin/sialophorin antigens.