Neutralising antibodies to the granulocyte colony-stimulating factor receptor recognise both the immunoglobulin-like domain and the cytokine receptor homologous domain.
To define regions of the granulocyte colony-stimulating factor (G-CSF) receptor that are important for ligand binding, neutralising monoclonal antibodies to the human receptor have been produced. Eleven antibodies recognised six different receptor epitopes. Antibodies from three of the epitope groups were able to detect the receptor by western blotting but did not inhibit G-CSF binding. The other three antibody groups inhibited G-CSF binding either completely (groups 1 and 2) or partially (group 3). All the antibodies inhibited proliferation of BA/F3 cells expressing the G-CSF receptor to varying extents. By using human-marine chimeric receptors, the binding sites of the antibodies were mapped to the immunoglobulin-like domain (groups 1 and 3), the cytokine receptor homologous domain (group 2) or the fibronectin type III domains (groups 4 to 6). These results show that the immunoglobulin-like and cytokine receptor homologous domains of the receptor are important for ligand binding and subsequent signalling.
Layton
J E
JE
Ludwig Institute for Cancer Research, Melbourne Tumour Biology Branch, Parkville, Australia.
Iaria
J
J
Nicholson
S E
SE
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D015153:Blotting, Western; D016466:CHO Cells; D002455:Cell Division; D006224:Cricetinae; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005434:Flow Cytometry; D006031:Glycosylation; D016179:Granulocyte Colony-Stimulating Factor / Q000378:metabolism; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D009500:Neutralization Tests; D011233:Precipitin Tests; D011485:Protein Binding; D016188:Receptors, Granulocyte Colony-Stimulating Factor / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D016188:Receptors, Granulocyte Colony-Stimulating Factor; D011993:Recombinant Fusion Proteins; D014415:Tunicamycin; D016179:Granulocyte Colony-Stimulating Factor