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IL 1 5及IL 1 5R阳性细胞在成人T淋巴细胞性白血病 (ATL)、多发性骨髓瘤及炎症性自身免疫性疾病病理过程中起着重要作用 .应用基因重组技术 ,构建、表达靶向于IL 1 5受体的两种融合蛋白 ,为研制特异的可消除IL 1 5R高表达细胞的导向药物奠定基础 .将人IL 1 5成熟肽基因及IL 1 5R拮抗剂 (IL 1 5M)基因片段分别与人工改造的人肿瘤坏死因子突变体 (TNFαM)基因按正确的阅读框架融合 ,定向克隆在pET1 6b表达载体T7启动子的下游 ,得到质粒pET IL 1 5 TNFαM和pET IL 1 5M TNFαM .从大肠杆菌相应重组菌株中通过Ni2 + NTA亲和层析分别纯化出两种融合蛋白IL 1 5 TNFαM和IL 1 5M TNFαM .IL 1 5 TNFαM和IL 1 5M TNFαM对IL 1 5R阳性红白血病细胞K5 6 2的杀伤作用分别是TNFα的 4和 1 5倍 ,两种蛋白对IL 1 5R阴性细胞系Jurkat的杀伤作用则没有明显差异且均弱于TNFα .这些结果说明 ,两种融合蛋白特别是IL 1 5受体拮抗型蛋白IL 1 5M TNFαM对与IL 1 5 IL 1 5R异常表达相关的疾病可能具有潜在的治疗价值
IL-15 and IL-15R-positive cells play an important role in the pathogenesis of adult T-cell lymphocytic leukemia (ATL), multiple myeloma and inflammatory autoimmune diseases.Using gene recombination technology, The two fusion proteins of IL-1 receptor and IL-5 receptor could lay the foundation for the development of a specific drug that can eliminate IL-5R-overexpressing cells.The human IL-15 mature peptide gene and IL15R antagonist (IL15M) gene fragment The human TNFR gene was fused with the human TNFαM gene in the correct reading frame and cloned downstream of the T7 promoter of pET1 6b expression vector to obtain the plasmids pET IL 1 5 TNFαM and pET IL 1 5M TNFαM. Two recombinant proteins IL15 TNFαM and IL15M TNFαM. IL15 TNFαM and IL15M TNFαM were purified from the corresponding recombinant strains of Escherichia coli by Ni2 + NTA affinity chromatography respectively. The results showed that IL-5R-positive erythroleukemia cells K5 6 2 were 4 and 15 folds of TNFα, respectively, and the killing effects of the two proteins on Jurkat, an IL 1 5R negative cell line, were not significantly different from that of TNFα. These results indicate that the two fusion proteins, especially IL 1 5 by Antagonistic IL 1 5M TNFαM protein associated disease may have potential therapeutic value for the abnormal expression of IL 1 5 IL 1 5R