Hannigan G E, Gewert D R, Williams B R
J Biol Chem. 1984 Aug 10;259(15):9456-60.
Interferon-sensitive (IFN-S) and IFN-resistant (IFN-R) Daudi lymphoblastoid cells were studied for IFN-alpha receptor expression and regulation by steady state and kinetic procedures, utilizing a homogeneous 125I-IFN-alpha 2 probe. Heterogeneity in the binding of this probe to IFN-S cells was determined to result from negatively cooperative interactions between an initially homogeneous class of IFN receptor. No such heterogeneity was noted in the IFN-R cells, indicating an apparent difference in the interaction of IFN-alpha 2 with these cells. The apparent dissociation constants (Kd) for IFN-S cell receptors were calculated to be 1 X 10(-10) M and 1 X 10(-8) M, for the high and low affinity sites, respectively. The Kd for sites on the IFN-R cells was estimated to be 4 X 10(-9) M. IFN-R and IFN-S cells expressed 2.4 X 10(4) and 3.5 X 10(4) binding sites per cell, respectively, representing an increase of at least 6-fold over previous reports of IFN-S Daudi IFN receptor density. Both IFN-S and IFN-R cells were capable of down-regulating expression of the IFN-alpha receptor in response to low concentrations of IFN-alpha 2. Furthermore, both cell lines were shown to be capable of internalizing specifically bound 125I-IFN-alpha 2 to an equivalent degree. Accordingly, we propose that the relative insensitivity of the Daudi IFN-R phenotype involves the loss of a high affinity interaction between cellular receptors and IFN-alpha 2, in addition to the reduced level of expressed low affinity binding sites.
利用均一的¹²⁵I - IFN-α₂探针,通过稳态和动力学方法研究了干扰素敏感(IFN - S)和干扰素抗性(IFN - R)的Daudi淋巴母细胞系中IFN - α受体的表达和调控。确定该探针与IFN - S细胞结合的异质性是由最初均一的一类IFN受体之间的负协同相互作用引起的。在IFN - R细胞中未观察到这种异质性,表明IFN - α₂与这些细胞相互作用存在明显差异。对于IFN - S细胞受体,高亲和力和低亲和力位点的表观解离常数(Kd)分别计算为1×10⁻¹⁰ M和1×10⁻⁸ M。IFN - R细胞上位点的Kd估计为4×10⁻⁹ M。IFN - R和IFN - S细胞分别每个细胞表达2.4×10⁴和3.5×10⁴个结合位点,比先前关于IFN - S Daudi IFN受体密度的报道增加了至少6倍。IFN - S和IFN - R细胞都能够响应低浓度的IFN - α₂下调IFN - α受体的表达。此外,两种细胞系都显示出能够同等程度地内化特异性结合的¹²⁵I - IFN - α₂。因此,我们提出Daudi IFN - R表型的相对不敏感性除了表达的低亲和力结合位点水平降低外,还涉及细胞受体与IFN - α₂之间高亲和力相互作用的丧失。