Benjamin D, Rosolen A, Wormsley S B, DeBault L E, Colamonici O R
Oncology Division, Saint Francis Research Institute, Oklahoma City, Oklahoma.
Cell Immunol. 1990 Aug;129(1):112-24. doi: 10.1016/0008-8749(90)90191-s.
IL-2 receptors on T cells exist in at least three forms which differ in their ligand-binding affinity. The low-affinity IL-2 receptor (IL-2R) consists of the 55-kDa Tac protein (p55 alpha), the intermediate-affinity site corresponds to the 70-kDa molecule (p70 beta), and the high-affinity IL-2R consists of a noncovalent heterodimeric structure involving both p55 alpha and p70 beta. We studied 24 B cell lines (8 EBV-negative and 16 EBV-positive) for IL-2R expression in the presence or absence of the tumor promoter, teleocidin. 125I-IL-2 radioreceptor binding assays and crosslinking studies demonstrated the sole expression of p55 alpha in EBV-negative cell lines only, whereas p55 alpha present in EBV-positive cell lines was always associated with p70 beta to construct high-affinity IL-2R. p70 beta was not detected in any of the EBV-negative cell lines, but was expressed on most of the EBV-positive cell lines (13 of 16). Our data also indicate that the expression of p55 alpha and p70 beta by radiolabeling correlates with their expression in flow cytometry, and that a large excess of p55 alpha is required to construct high-affinity IL-2R. Coexpression of p55 alpha and p70 beta on human B cells contributed to constructing high-affinity IL-2R hybrid complex as shown by (i) rapid association rate contributed by p55 alpha and slow dissociation rate by p70 beta; (ii) teleocidin's ability to induce p55 alpha on cell lines which express p70 beta only, resulting in appearance of high-affinity IL-2R; (iii) blocking p55 alpha by anti-Tac mAb in cell lines which constitutively express high-affinity IL-2R eliminated both high- and low-affinity components. The existence of low, intermediate, and high IL-2R on human B cells bears important future implications for understanding the mechanism of IL-2 signaling and the role of IL-2 in B cell activation, proliferation, and differentiation.
T细胞上的白细胞介素-2(IL-2)受体至少以三种形式存在,它们的配体结合亲和力各不相同。低亲和力IL-2受体(IL-2R)由55 kDa的Tac蛋白(p55α)组成,中等亲和力位点对应于70 kDa的分子(p70β),高亲和力IL-2R由涉及p55α和p70β的非共价异二聚体结构组成。我们研究了24个B细胞系(8个EBV阴性和16个EBV阳性)在存在或不存在肿瘤启动子远藤菌素的情况下IL-2R的表达。125I-IL-2放射受体结合试验和交联研究表明,p55α仅在EBV阴性细胞系中单独表达,而EBV阳性细胞系中存在的p55α总是与p70β结合以构建高亲和力IL-2R。在任何EBV阴性细胞系中均未检测到p70β,但在大多数EBV阳性细胞系(16个中的13个)中表达。我们的数据还表明,通过放射性标记检测到的p55α和p70β的表达与其在流式细胞术中的表达相关,并且构建高亲和力IL-2R需要大量过量的p55α。如以下所示,人B细胞上p55α和p70β的共表达有助于构建高亲和力IL-2R杂合复合物:(i)p55α促成快速缔合速率,p70β促成缓慢解离速率;(ii)远藤菌素诱导仅表达p70β的细胞系上p55α的能力,导致高亲和力IL-2R出现;(iii)在组成性表达高亲和力IL-2R的细胞系中用抗Tac单克隆抗体阻断p55α消除了高亲和力和低亲和力成分。人B细胞上低、中和高亲和力IL-2R的存在对于理解IL-2信号传导机制以及IL-2在B细胞活化、增殖和分化中的作用具有重要的未来意义。