Development of IL-1 with selective activities by introduction of carbohydrates

IL-1具有抗肿瘤作用，对动物具有抗感染、抗辐射、抗化疗等保护作用，具有潜在的治疗价值。然而，IL-1表现出严重的有害作用。在这项研究中，我们试图通过在重组人IL-1α中化学引入低聚糖来开发危害较小的IL-1。1)甘露糖二聚体引入的IL-1的组织分布以前我们已经成功地合成了甘露糖二聚体引入的具有体内选择性活性的IL-1α。在这项研究中，检测了糖基化对组织分布的影响。将甘露糖二聚体引入的IL-1标记为~(125)&gt；I，然后观察其在小鼠体内的组织分布。与未糖化的IL-1相比，糖基化的IL-1在肝脏中的分布更多，在肾脏中的分布更少。2)甘露糖二聚体引入的IL-1糖基化位点的分析用糖内肽酶和糖基化…TOF-MASS分析了更多的d位点，IL-1α和IL-1β之间的保守区没有糖基化。3)半乳糖引入的IL-1D-半乳糖单糖的合成、纯化和生物活性通过酰化叠氮法偶联到IL-1α上，并用阴离子交换柱层析进行纯化。在体外，我们检测了多种IL-1活性，包括对T细胞的增殖作用，对髓系白血病细胞和黑色素瘤细胞的抗增殖作用，对黑色素瘤细胞合成IL-6的刺激作用，对成纤维细胞合成前列腺素E_2的刺激作用。在体外实验中，糖基化的IL-1的活性比非糖基化的IL-1的活性降低了10~1000倍。糖基化的IL-1与I型和II型IL-1受体的结合亲和力降低，这表明这种活性的降低至少部分是由于结合亲和力的降低。较少

IL-1 is potentially useful for therapy because of its antitumor effect and protective effect on animals against infection, radiation and chemotherapy. However, IL-1 exhibits serious deleterious effects. In this study we attempted to develop IL-1 with less deleterious effects by chemically introducing oligosaccharides to recombinant human IL-1alpha.1) Tissue distribution of mannose dimer-introduced IL-1We have previously succeeded in the synthesis of mannose dimer-introduced IL-1alpha with selective activities in vivo. In this study the effect of glycosylation on tissue distribution was examined. Mannose dimer-introduced IL-1 was labeled with ^<125>I,and then examined its tissue distribution after i.p.or i.v.injection into mice. Compared to the nonglycosylated IL-1, the glycosylated IL-1 distributed more in the liver, and less in the kidney.2) Analysis of glycosylated site in mannose dimer-introduced IL-1Mannose dimer-introduced IL-1 was dijested with lysylendopeptidase, and glycosylate … More d sites were analyzed by TOF-MASS.The conserved regions among IL-1alpha and IL-1beta were not glycosylated. However, the regions exposed to the surface of IL-1alpha molecule were glycosylated.3) Synthesis, purification and biological activities of galactose-introduced IL-1D-Galactose monosaccharide was coupled to IL-1alpha by the acyl azide method, and was purified by anion-exchange column chromatography. Galactose introduced per molecule of IL-1 was estimated to be 9. We examined a variety of IL-1 activities in vitro, including proliferative effect on T cells, antiproliferative effect on myeloid leukemia cells and melanoma cells, stimulatory effects on IL-6 syntheisi by melanoma cells and PGE2 synthesis by fibroblast cells. The glycosylated IL-1 exhibited reduced activities from ten to thousand times compared with nonglycosyslated IL-1 in all the activities performed in vitro. The glycosylated IL-1 exhibited reduced binding affinities to type I and II IL-1 receptors, suggesting that the reduced activities are due, at least partially, to the decrease in the binding affinities. Less