An 80,000-kd glycoprotein cell surface antigen found only on nonhematopoietic cells in human bone marrow.
The murine IgG2a monoclonal antibody (MoAb) 6-19 binds to the surface of human nonhematopoietic cells but not to hematopoietic cells. As previously described, it can be used with complement to selectively kill nonhematopoietic cells prior to culture of human bone marrow. It is now shown that the 6-19 MoAb recognizes a specific antigenic determinant of apparent molecular mass 80 kd, detected by western blotting on nonhematopoietic tumor cell lines, endothelial cells, and bone marrow stromal cells. Mouse L cells were transfected with human DNA, and the 6-19 monoclonal antibody selected cells that expressed an antigen with similar characteristics. The antigenic determinant is absent in hematopoietic tumor cell lines, peripheral blood cells, and bone marrow hematopoietic cells. Investigation of the biochemical nature of the antigen, using various enzymes, lectin-binding studies, and western blotting suggests that the 6-19 epitope is at least partially carbohydrate and that the protein has N-linked sugars but not O-linked. The affinity (Ka approximately 10(9) M-1) of MoAb 6-19 binding is similar in tumor cells and normal fibroblasts and endothelial cells. The identification of a specific antigenic determinant of 80 kd may help to discriminate between hematopoietic and nonhematopoietic cells in human bone marrow.
Iyer
J
J
University of Rochester Medical Center, NY 14642.
Duerst
R E
RE
Looney
J N
JN
Humphries
R K
RK
Abboud
C N
CN
Frantz
C N
CN
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis* / Q000235:genetics; D015153:Blotting, Western; D001853:Bone Marrow / Q000276:immunology*; D001854:Bone Marrow Cells; D002460:Cell Line; D004247:DNA / Q000235:genetics; D000939:Epitopes / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis* / Q000235:genetics; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000276:immunology; D006801:Humans; D007163:Immunosorbent Techniques; D007739:L Cells; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D051379:Mice; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D000939:Epitopes; D006023:Glycoproteins; D014415:Tunicamycin; D004247:DNA; D006026:Glycoside Hydrolases; D009439:Neuraminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase