Biologic properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth factor activity, mast cell growth factor activity, p cell-stimulating factor activity, colony-stimulating factor activity, and histamine-producing cell-stimulating factor activity.
Biologic properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth factor activity, mast cell growth factor activity, p cell-stimulating factor activity, colony-stimulating factor activity, and histamine-producing cell-stimulating factor activity.
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DOI：
10.4049/jimmunol.131.5.2609.c
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发表时间：
1983-07
影响因子：
4.4
通讯作者：
J. Ihle;J. Keller;S. Oroszlan;L. Henderson;T. Copeland;F. Fitch;M. Prystowsky;E. Goldwasser;J. Schrader;E. Palaszynski;M. Dy;B. Lebel
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文献类型：
--
作者：
J. Ihle;J. Keller;S. Oroszlan;L. Henderson;T. Copeland;F. Fitch;M. Prystowsky;E. Goldwasser;J. Schrader;E. Palaszynski;M. Dy;B. Lebel
白细胞介素 3 (IL 3) 最初被定义为来自刀豆球蛋白 A 刺激的淋巴细胞 (Con A CM) 的条件培养基中的一种因子，可在 nu/nu 脾淋巴细胞培养物中诱导 20-α-羟基类固醇脱氢酶 (20 α SDH)。为了确定其他“生物”活性的范围，将 IL 3 纯化至同质并评估其特性。根据以下标准判断蛋白质制剂为均质 IL 3：1) 在纯化的最后一步中以恒定比活性洗脱 IL 3 峰，2) 通过 SDS-PAGE 分析存在单一蛋白质，3) 针对 IL 3 依赖性细胞系的受体结合活性，4) 比活性与 50% 最大生物活性所需的 0.2 ng/ml 一致，以及 5) 存在单一氨基末端序列。使用这种 IL 3 制剂，20 α SDH 诱导的剂量反应曲线与 1) WEHI-3 生长因子、2) 肥大细胞生长因子、3) P 细胞刺激因子和 4) 组胺产生细胞刺激因子活性的剂量反应曲线相同或相似。此外，同质 IL 3 具有集落刺激因子活性，尽管 Con A CM 中发现的总 CSF 活性中只有大约 2% 与 IL 3 相关。CSF 活性的主峰可以通过 DEAE 柱色谱从 IL 3 中分离出来，并且缺乏许多与 IL 3 相关的生物活性。
Interleukin 3 (IL 3) was initially defined as a factor in conditioned media from concanavalin A-stimulated lymphocytes (Con A CM) that induces the enzyme 20-alpha-hydroxysteroid dehydrogenase (20 alpha SDH) in cultures of nu/nu splenic lymphocytes. To determine the spectrum of additional "biologic" activities, IL 3 was purified to homogeneity and its properties were assessed. The protein preparation was judged to be homogeneous IL 3 by the following criteria: 1) elution of a peak of IL 3 with a constant specific activity in the last step of purification, 2) presence of a single protein by SDS-PAGE analysis, 3) receptor-binding activity against IL 3-dependent cell lines, 4) a specific activity of congruent to 0.2 ng/ml required for 50% of maximal biologic activity, and 5) the presence of a single amino terminal sequence. With the use of this preparation of IL 3, the dose-response curves for 20 alpha SDH induction were identical or similar to the dose-response curves for the activities of 1) WEHI-3 growth factor, 2) mast cell growth factor, 3) P cell-stimulating factor, and 4) histamine-producing cell-stimulating factor. In addition, homogeneous IL 3 had colony-stimulating factor activity, although only approximately 2% of the total CSF activity found in Con A CM was associated with IL 3. The major peak of CSF activity could be resolved from IL 3 by DEAE column chromatography and lacked many of the biologic activities associated with IL 3.