A monoclonal antibody with IL-3-like activity blocks IL-3 binding and stimulates tyrosine phosphorylation.
IL-3 is a growth factor for multi-potential hemopoietic cells. A panel of mAb with IL-3-like activities was recently derived from the autoimmune mouse MRL/1 pr. We present detailed evidence that one of these monoclonal antibodies, M7B1-5.1-F9 (F9), interacts with the mouse IL-3 receptor or with part of an IL-3R complex. F9 is a full agonist (80 to 100%) of IL-3 in proliferation assays, with a half-maximum effective concentration (EC50) of 0.2 to 2.0 nM. However, in the variant cell line, NFS60.8, the EC50 for F9 is 30 nM. The decreased sensitivity to the antibody is also paralleled by an increased requirement (EC50) for IL-3. Two stromal cell lines also show increased requirements for IL-3 and F9. F9 stimulates the tyrosine phosphorylation of the same set of proteins phosphorylated after IL-3 interaction with the IL-3R, suggesting that IL-3 and F9 activate the same tyrosine kinase. F9 specifically inhibits 125I-IL-3 binding at a concentration (IC50) of about 300 nM, two log10 orders of magnitude higher than that required for its agonistic effects, suggesting that spare receptors may exist. In cross-linking assays, F9 blocks the specific binding of 125I-IL-3 to proteins of Mr 140, 130, and 70 kDa. Thus, F9 interacts with the IL-3R at or near the binding site, which leads to the stimulation of a tyrosine kinase and cell proliferation.
Schreurs
J
J
Department of Molecular Biology, DNAX Research Institute, Palo Alto, CA 94304.
Sugawara
M
M
Arai
K
K
Ohta
Y
Y
Miyajima
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002460:Cell Line; D003432:Cross-Linking Reagents; D007377:Interleukin-3 / Q000378:metabolism* / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010766:Phosphorylation; D011971:Receptors, Immunologic / Q000276:immunology; D016185:Receptors, Interleukin-3; D014443:Tyrosine / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D007377:Interleukin-3; D011971:Receptors, Immunologic; D016185:Receptors, Interleukin-3; D014443:Tyrosine