Selection of BoCD25 monoclonal antibodies by screening mouse L cells transfected with the bovine p55-interleukin-2 (IL-2) receptor gene.
The bovine interleukin-2 receptor-alpha (IL-2R alpha) gene has been isolated and a rabbit antiserum against a fusion protein of the gene has been produced. However, the antiserum does not inhibit IL-2-dependent proliferation. Since a panel of monoclonal antibodies (mAb) to bovine activation antigens was available, we transfected the gene into mouse L fibroblasts, selected stable transfectants with the rabbit antiserum, and screened for antibodies that bound the transfected cells but not the untransfected cells. Three mAb were selected and all three precipitated a molecule of M(r) 55,000 (under reducing conditions) from activated cells, as expected from homology with mouse and human IL-2R alpha (CD25, Tac). One of the three mAb was a strong inhibitor of IL-2-dependent proliferation of bovine lymphocytes. Thus, the availability of transfected cells allowed us to establish quickly and unequivocally the antigenic specificity of a number of antibodies.
Naessens
J
J
ILRAD, Nairobi, Kenya.
Sileghem
M
M
MacHugh
N
N
Park
Y H
YH
Davis
W C
WC
Toye
P
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002417:Cattle / Q000276:immunology*; D002455:Cell Division / Q000276:immunology; D002460:Cell Line; D005796:Genes / Q000276:immunology*; D007376:Interleukin-2 / Q000276:immunology; D051379:Mice; D015375:Receptors, Interleukin-2 / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D014162:Transfection / Q000276:immunology
D000911:Antibodies, Monoclonal; D007376:Interleukin-2; D015375:Receptors, Interleukin-2