Studies on the molecular nature of human interleukin 1.
Studies on the molecular nature of human interleukin 1.
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DOI：
10.4049/jimmunol.138.5.1447
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发表时间：
1987-03
影响因子：
4.4
通讯作者：
P. Auron;S. Warner;A. C. Webb;J. Cannon;H. Bernheim;K. McAdam;L. Rosenwasser;G. LoPreste;S. F. Mucci;C. Dinarello
中科院分区：
文献类型：
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作者：
P. Auron;S. Warner;A. C. Webb;J. Cannon;H. Bernheim;K. McAdam;L. Rosenwasser;G. LoPreste;S. F. Mucci;C. Dinarello
在35s -蛋氨酸-、[3H]亮氨酸-或14c标记的氨基酸存在的情况下，用热杀死的白色葡萄球菌或大肠杆菌脂多糖刺激贴壁的人血液单核细胞。孵育后，用抗人白细胞介素1免疫吸附剂纯化上清培养基中的白细胞介素1 （IL 1）活性，然后进行凝胶过滤和层析聚焦。通过单、二维sds -聚丙烯酰胺凝胶电泳的荧光图评估IL - 1的纯度。对低毫瓦蛋白质（小于20,000毫瓦）的等电和染色质聚焦发现了三种带电荷的18,000毫瓦的IL - 1，其pI值约为7、6和5，其中在pI 7处最丰富。在纯化过程中，淋巴细胞共丝分裂活性、家兔发热和真皮成纤维细胞的前列腺素E2释放在相同馏分中共洗脱。此外，当注射到内毒素抗性C3H/HeJ小鼠体内时，这些组分具有产生发热、诱导血清淀粉样蛋白A、降低血清铁浓度和增加循环中性粒细胞数量的活性。荧光成像显示均匀的条带，m.w.约为18,000，与这些生物活性相关。根据T细胞共丝分裂活性与合并的放射性标记氨基酸的比值来判断，pI 6或5 IL 1的比活性至少比pI 7形式高10倍。这一结果表明，这两种18000毫瓦特酸性形态的氨基酸组成与pI - 7物种无关。这些结果还表明，pI - 7人类单核细胞IL - 1是主要的18000 m.w.合成形式，此外，均质pI - 7 IL - 1通过调节免疫、炎症、代谢和神经功能在各种组织中表现出多种生物学特性。数据也提出了存在高mw（32,000）的人前il - 1分子作为主要的单核细胞内形式。在细胞外丝氨酸蛋白酶活性存在的情况下，这种il - 1在分离过程中被人为地降解为低分子量形式。这些数据与IL - 1分泌模型一致，在该模型中，IL - 1前体首先在细胞内合成，并在细胞外运输期间或之后进行加工。
Adherent human blood monocytes were stimulated with heat-killed Staphylococcus albus or Escherichia coli lipopolysaccharide in the presence of 35S-methionine-, [3H]leucine-, or 14C-labeled amino acids. After incubation, interleukin 1 (IL 1) activity in the supernatant medium was purified over an anti-human IL 1 immunoadsorbent followed by gel filtration and chromatofocusing. The purity of the IL 1 was assessed by fluorography of one- and two-dimensional SDS-polyacrylamide gel electrophoresis. Isoelectric and chromatofocusing of low m.w. proteins (less than 20,000 m.w.) revealed three charged 18,000 m.w. species of IL 1 with approximate pI's of 7, 6, and 5, with the most abundant form at pI 7. During the purification procedures, lymphocyte co-mitogenic activity, fever in rabbits, and prostaglandin E2 release from dermal fibroblasts co-eluted in the same fractions. In addition, these fractions were active when injected into endotoxin-resistant C3H/HeJ mice for the production of fever, the induction of serum amyloid A protein, a decrease in serum iron concentration, and an increase in the number of circulating neutrophils. Fluorography revealed homogeneous bands with an m.w. of about 18,000 which correlated with these biological activities. The specific activity of the pI 6 or 5 IL 1, as judged by the ratio of T cell co-mitogenic activity to incorporated radiolabeled amino acid, was at least 10-fold greater than that observed for the pI 7 form. This result suggests that the amino acid compositions of the two 18,000 m.w. acidic forms are unrelated to the pI 7 species. These results also demonstrate that the pI 7 human monocyte IL 1 is the predominant 18,000 m.w. form synthesized and, furthermore, that homogeneous pI 7 IL 1 exhibits multiple biological properties on various tissues by modulating immunologic, inflammatory, metabolic, and neurologic functions. Data are also presented for the existence of a high m.w. (32,000) human pro-IL 1 molecule as the predominant monocytic intracellular form. This pro-IL 1 is degraded artifactually during isolation to lower m.w. forms in the presence of an extracellular serine protease activity. These data are consistent with a model for IL 1 secretion in which pro-IL 1 is first synthesized within the cell and is processed during or after extracellular transport.