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同源白细胞介素3的生物学特性。I. WEHI-3生长因子活性、肥大细胞生长因子活性、β细胞刺激因子活性、集落刺激因子活性及组胺产生细胞刺激因子活性的证明。

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.

作者信息

Ihle J N, Keller J, Oroszlan S, Henderson L E, Copeland T D, Fitch F, Prystowsky M B, Goldwasser E, Schrader J W, Palaszynski E, Dy M, Lebel B

出版信息

J Immunol. 1983 Jul;131(1):282-7.

PMID:6190911
Abstract

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.

摘要

白细胞介素3(IL-3)最初被定义为刀豆球蛋白A刺激的淋巴细胞(Con A CM)条件培养基中的一种因子,它能在裸鼠脾脏淋巴细胞培养物中诱导20-α-羟基类固醇脱氢酶(20α SDH)。为了确定其他“生物学”活性的范围,将IL-3纯化至同质,并评估其特性。通过以下标准判断该蛋白质制剂为同质IL-3:1)在纯化的最后一步中,IL-3峰以恒定的比活性洗脱;2)SDS-PAGE分析显示存在单一蛋白质;3)对IL-3依赖性细胞系具有受体结合活性;4)最大生物学活性的50%所需的比活性约为0.2 ng/ml;5)存在单一的氨基末端序列。使用这种IL-3制剂,20α SDH诱导的剂量反应曲线与以下活性的剂量反应曲线相同或相似:1)WEHI-3生长因子;2)肥大细胞生长因子;3)P细胞刺激因子;4)组胺产生细胞刺激因子。此外,同质IL-3具有集落刺激因子活性,尽管在Con A CM中发现的总CSF活性中只有约2%与IL-3相关。CSF活性的主要峰可以通过DEAE柱色谱与IL-3分离,并且缺乏许多与IL-3相关的生物学活性。

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