Characterization of a high affinity interleukin-1 (IL-1) specific binding site in a human synovial sarcoma (Hs431) cell line.

Lymphokine research Pub Date : 1990-01-01
D End, T Garrabrant
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Abstract

A subpopulation of cells was derived from the Hs431 connective tissue sarcoma cell line which possessed high affinity (estimated Kd = 0.38-0.55 nM) binding sites for human recombinant [125I]-IL-1 alpha. Binding at 4 degrees C was slow approaching equilibrium by 4 hrs. Dissociation of [125I]-IL-1 alpha was also slow and unaffected by high concentrations of cold ligand. The binding site also underwent ligand-induced internalization at 37 degrees C. An Mr = 83,000 protein was identified in affinity crosslinking studies. Despite these similarities to previously reported IL-1 receptors, Hs431 cells did not exhibit biological responses to IL-1 which have been observed in other cell lines. IL-1 did not induce PGE2 or collagenase synthesis. IL-1 also failed to induce ornithine decarboxylase activity (ODC) or stimulate [3H]-thymidine incorporation. In contrast, the Hs431 cells did contain a functional epidermal growth factor (EGF) receptor as determined from binding studies, protein kinase activity, induction of ODC, and stimulation of [3H]-thymidine incorporation. Thus, the refractoriness of Hs431 cells to IL-1 was fairly specific and did not result from a generalized defect associated with cell transformation.

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人滑膜肉瘤(Hs431)细胞系中高亲和力白细胞介素-1 (IL-1)特异性结合位点的表征
从Hs431结缔组织肉瘤细胞系中获得了具有高亲和力(估计Kd = 0.38-0.55 nM)的人重组[125I]-IL-1 α结合位点的细胞亚群。在4℃下结合缓慢接近平衡4小时。[125I]-IL-1 α的解离也很慢,不受高浓度冷配体的影响。结合位点在37℃下也经历了配体诱导的内化,在亲和交联研究中鉴定出Mr = 83,000的蛋白。尽管与先前报道的IL-1受体有这些相似之处,但Hs431细胞并未表现出在其他细胞系中观察到的对IL-1的生物学反应。IL-1不诱导PGE2或胶原酶合成。IL-1也不能诱导鸟氨酸脱羧酶活性(ODC)或刺激[3H]-胸苷结合。相比之下,通过结合研究、蛋白激酶活性、ODC诱导和[3H]-胸腺嘧啶结合刺激确定,Hs431细胞确实含有功能性表皮生长因子(EGF)受体。因此,Hs431细胞对IL-1的难耐性是相当特异性的,并不是由于与细胞转化相关的普遍缺陷。
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