Characterization and use of monoclonal and polyclonal antibodies against the mouse interferon-gamma receptor.
To facilitate investigation of its physical and functional properties, 11 monoclonal antibodies (mAbs) and a goat polyclonal IgG specific for the mouse interferon- (IFN-gamma) receptor were characterized and their potential uses studied. Eight of the mAbs interacted with epitopes on the extracellular domain of the receptor, two interacted with epitopes on the intracellular domain, and one interacted with an epitope that could not be localized definitively to either region. Of the 11 mAbs, the majority (8) were IgGs, 2 were IgMs, and 1 was an IgA. Relative avidities of the seven that could be determined ranged from 333 to 0.002 microM-1. Both the polyclonal goat IgG and mAb GR-20 (the latter specific for an epitope in the binding site for IFN-gamma) blocked binding of the ligand and, as expected, prevented induction by IFN-gamma of priming of macrophages for tumor cell killing. None of the other mAbs had an effect despite the fact that GR-22 partially (greater than 50%) blocked binding of IFN-gamma. Neither the polyclonal IgG nor any of the mAbs had an agonist effect. The relative usefulness of the antibodies for immunoprecipitation, immunoblotting, immunoassay, and cell staining with and without prior fixation is described. The results of immunocytochemical staining directly confirmed that the majority of immunologically reactive receptor protein expressed by cells is intracellular. To facilitate use by other investigators, the hybridomas that produce these mAbs will be offered to the American Type Culture Collection.
LeClaire
R D
RD
Wilkinson Laboratory of the Cancer Center, University of Kansas School of Medicine, Kansas City.
Basu
M
M
Pinson
D M
DM
Redick
M L
ML
Hunt
J S
JS
Zavodny
P J
PJ
Pace
J L
JL
Russell
S W
SW
D000818:Animals; D000906:Antibodies / Q000145:classification; D000911:Antibodies, Monoclonal / Q000145:classification; D000918:Antibody Specificity; D015153:Blotting, Western; D002478:Cells, Cultured; D000939:Epitopes / Q000032:analysis; D005434:Flow Cytometry; D007150:Immunohistochemistry; D007371:Interferon-gamma / Q000032:analysis; D008024:Ligands; D051379:Mice; D008809:Mice, Inbred C3H; D011233:Precipitin Tests; D051381:Rats; D011971:Receptors, Immunologic / Q000737:chemistry* / Q000502:physiology; D017471:Receptors, Interferon; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D008024:Ligands; D011971:Receptors, Immunologic; D017471:Receptors, Interferon; D007371:Interferon-gamma