A monoclonal antibody identifies a 215 000-dalton nuclear envelope protein restricted to certain cell types.
The monoclonal antibody, AGF2.3, was isolated from mice immunised with the human promyeloid cell line HL60. By immunofluorescence and immunoelectron microscopy the antibody was shown to bind to the nuclear envelope in uninduced HL60 cells. Immunofluorescent staining was reduced to very low levels in HL60 cells induced to mature to monocytes or neutrophils by addition of 12-0-tetradecanoylphorbol-13-acetate or dimethyl sulfoxide respectively. Blood neutrophils did not express the antigen. Weak immunofluorescent staining of cell nuclei was observed in peripheral blood lymphocytes and in sections of normal human kidney, tonsil and skin epithelium. The AGF2.3 antigen was strongly expressed on the nuclei of 21/21 haemopoietic cell lines and 21/25 permanent non-haemopoietic cell lines representing various cell types. In contrast, the antigen was not expressed by any of six primary (untransformed) cell cultures. These included fibroblasts, endothelial cells and keratinocytes. The antigen was expressed in the Q10 SV-40 transformed cell line derived from a non-expressing primary fibroblast culture. AGF2.3 antibody precipitated a protein with an apparent subunit molecular weight of approximately 215 kDa from Triton X-100 extracts of HL60 and HeLa cells labelled with 35S-methionine. This protein was not detectable in extracts of primary skin fibroblasts prepared in parallel. We conclude that AGF2.3 antibody recognises a previously undescribed protein associated with the nuclear envelope which is expressed at high levels in most transformed cell lines but which is weakly expressed or absent in normal tissues and primary cell cultures.
Brown
G
G
Turner
B M
BM
Morris
C J
CJ
Bahman
A M
AM
Fisher
A G
AG
Whitfield
W G
WG
Davies
S
S
Barthakur
R
R
Johnson
G D
GD
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000032:analysis*; D000951:Antigens, Neoplasm / Q000032:analysis / Q000276:immunology; D034961:Antigens, Nuclear; D001324:Autoantigens / Q000032:analysis* / Q000276:immunology; D002460:Cell Line; D006412:Hematopoietic Stem Cells / Q000032:analysis / Q000276:immunology; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000276:immunology*; D051379:Mice; D008854:Microscopy, Electron; D008970:Molecular Weight; D009698:Nucleoproteins / Q000032:analysis* / Q000276:immunology; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D000941:Antigens; D000951:Antigens, Neoplasm; D034961:Antigens, Nuclear; D001324:Autoantigens; D009698:Nucleoproteins