Huang J J, Newton R C, Rutledge S J, Horuk R, Matthew J B, Covington M, Lin Y
Medical Products Department, E.I. du Pont de Nemours & Company, Glenolden, PA 19036.
J Immunol. 1988 Jun 1;140(11):3838-43.
One cDNA clone encoding a truncated murine IL-1 beta (M IL-1 beta) sequence was isolated from a murine macrophage cDNA library. We reconstituted the coding sequence of the 152-residue mature protein and expressed it in Escherichia coli. rM IL-1 beta was purified to homogeneity and characterized by oligonucleotide and NH2-terminal sequence analysis. Purified rM IL-1 beta exhibited biologic activity equivalent to 7.8 x 10(7) units/mg in the murine thymocyte proliferation assay and 9.9 x 10(3) units/mg in the human gingival fibroblast PGE2 production assay, indicative of species specificity. The isoelectric point of rM IL-1 was found to be 8.85. The circular dichroism spectrum revealed that the secondary structure of M IL-1 is indistinguishable from that of the human protein. Receptor binding studies indicated the rM IL-1 bound to murine EL-4.1 thymoma cells in a specific and dose-dependent fashion with an affinity of 32 pM. Competition binding data suggested that murine and human IL-1 compete for a single class of receptor. Antisera were generated in rabbits against both murine and human IL-1. Results of ELISA binding and antisera neutralization assays indicated that there are common antigenic sites between the two IL-1 beta molecules. These domains are of functional importance because they are capable of mediating the neutralization of biologic activity.
从鼠巨噬细胞cDNA文库中分离出一个编码截短型鼠白细胞介素-1β（M IL-1β）序列的cDNA克隆。我们重构了152个氨基酸残基的成熟蛋白的编码序列，并在大肠杆菌中进行表达。重组鼠白细胞介素-1β（rM IL-1β）被纯化至同质，并通过寡核苷酸和氨基末端序列分析进行表征。在鼠胸腺细胞增殖试验中，纯化的rM IL-1β表现出相当于7.8×10⁷单位/毫克的生物活性，在人牙龈成纤维细胞前列腺素E2产生试验中为9.9×10³单位/毫克，表明具有种属特异性。发现rM IL-1的等电点为8.85。圆二色光谱显示，M IL-1的二级结构与人类蛋白的二级结构无法区分。受体结合研究表明，rM IL-1以特异性和剂量依赖性方式与鼠EL-4.1胸腺瘤细胞结合，亲和力为32 pM。竞争结合数据表明，鼠和人白细胞介素-1竞争同一类受体。用兔制备了针对鼠和人白细胞介素-1的抗血清。酶联免疫吸附测定（ELISA）结合和抗血清中和试验结果表明，两种白细胞介素-1β分子之间存在共同的抗原位点。这些结构域具有功能重要性，因为它们能够介导生物活性的中和。
