Parameters of interaction of a novel monoclonal antibody (33B3.1) with the human IL2-receptors: interrelationship between 33B3.1, anti-Tac, and IL2 binding sites.
This report describes the molecular parameters of interaction of a new antibody (33B3.1) with the human membrane RIL2 expressed by ConA-activated T lymphocytes or allogeneic T-cell clones established from a rejected kidney allograft: the 33B3.1 immunoprecipitates a membrane protein of 55000 MW. It inhibits IL2-driven proliferation of activated T cells. This inhibition occurred in the nanomolar range when low concentrations of recombinant IL2 (rec-IL2) were used. The (125I)-33B3.1 binds in a specific way to a single class of receptor sites on activated T cells. The rate constants of association and dissociation at 37 degrees C of the labeled 33B3.1 were k*1 = 12 X 10(5) M-1 s-1 and k*-1 = 7 X 10(-4) s-1, respectively, and its equilibrium dissociation constant was KD = 0.65 nM. Maximal binding capacities were fairly variable among T-cell clones, as high as 300,000 sites/cell for some of them. Competition experiments demonstrate that the 33B3.1 and anti-Tac interact with the RIL2 in a competitive manner, suggesting that they recognize closely associated epitopes on the RIL2. However, the 33B3.1 inhibits the binding of (35S)-recombinant IL2 to its high affinity RIL2 in a noncompetitive way. The 33B3.1 seems therefore to interact with an epitope close but distinct from the IL2 binding site. Our data could suggest either that the 33B3.1 is able to convert high affinity RIL2 towards low affinity conformations or that there is more than one IL2 binding site per molecule of high affinity RIL2.
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D001665:Binding Sites; D001667:Binding, Competitive; D002999:Clone Cells / Q000276:immunology; D006801:Humans; D008213:Lymphocyte Activation; D011971:Receptors, Immunologic / Q000276:immunology*; D015375:Receptors, Interleukin-2; D013601:T-Lymphocytes / Q000276:immunology; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7