Characterization of the soluble murine IL-2R and estimation of its affinity for IL-2.
IL-2 induces cells of the cytotoxic T cell line C30.1 to express large numbers of membrane IL-2R (mIL-2R). At the height of activation, these cells also release a soluble form of IL-2R (sIL-2R). Using either crude supernatant or a semi-purified preparation of sIL-2R obtained by affinity chromatography, studies were performed to characterize murine sIL-2R. Its m.w. was determined by both gel filtration and SDS-PAGE. The affinity of sIL-2R for a panel of mAb known to recognize different epitopes of mIL-2R (p55 subunit) was assessed by saturation and competition experiments. The relationship between the various epitopes was studied by cross-inhibition experiments. The data suggest that sIL-2R and mIL-2R (p55 subunit) are structurally similar. The ability of sIL-2R to bind IL-2 was assessed by measuring the dissociation and the inhibition constant of the molecule for IL-2. Both values coincide and indicate that the affinity of sIL-2R for IL-2 is at least 10-fold lower than the that of low affinity mIL-2R. The biologic implications of these findings are discussed.
D000818:Animals; D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002460:Cell Line; D002474:Cell-Free System; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007376:Interleukin-2 / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008970:Molecular Weight; D011971:Receptors, Immunologic / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D015375:Receptors, Interleukin-2; D012967:Sodium Dodecyl Sulfate; D012995:Solubility
D000911:Antibodies, Monoclonal; D007376:Interleukin-2; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; D012967:Sodium Dodecyl Sulfate