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人辅助性T细胞因子(ThF)。I. 部分纯化与特性鉴定。

Human helper T cell factor(s) (ThF). I. Partial purification and characterization.

作者信息

Hirano T, Teranishi T, Toba H, Sakaguchi N, Fukukawa T, Tsuyuguchi I

出版信息

J Immunol. 1981 Feb;126(2):517-22.

PMID:6161177
Abstract

T cells obtained from pleural effusion of patients with tuberculous pleurisy were stimulated in vitro with PPD. The culture supernatant was shown to be substituted for T cells in PWM-induced IgG production by human B cells. As many as 0.5 to 3 x 10(9) lymphocytes were obtained from 1 patient, and about 85% of them were E-rosette positive, making it possible to purify human helper T cell factor(s) (ThF). Helper activity was sensitive to heating at 70 degrees C for 5 min and treatment with trypsin. ThF did not have conventional Ig determinants and was recovered in the 50 to 67% ammonium sulfate fraction. The purification was done by successive DEAE-Sephadex and CM-Sephadex ion-exchange chromatography, Sephadex G-100 gel filtration, and isoelectric focusing. It was shown that ThF activity was eluted by gel filtration in the fraction with a m.w. of about 20,000. Isoelectric focusing of this fraction revealed that ThF activity distributed in the pl range of 6.5 to 8.0, whereas TCGF activity was focused in a single peak at pI of 6.5, indicating the existence of a helper factor without TCGF activity.

摘要

从结核性胸膜炎患者胸腔积液中获取的T细胞,在体外以结核菌素纯蛋白衍生物(PPD)刺激。结果显示,该培养上清液可替代T细胞,用于人B细胞在美洲商陆有丝分裂原(PWM)诱导下产生IgG。从1例患者可获得多达0.5至3×10⁹个淋巴细胞,其中约85%为E花环阳性,这使得纯化人辅助性T细胞因子(ThF)成为可能。辅助活性对70℃加热5分钟及胰蛋白酶处理敏感。ThF没有传统的Ig决定簇,在50%至67%硫酸铵组分中可回收。通过连续的二乙氨基乙基葡聚糖(DEAE - Sephadex)和羧甲基葡聚糖(CM - Sephadex)离子交换层析、葡聚糖G - 100凝胶过滤及等电聚焦进行纯化。结果表明,通过凝胶过滤,ThF活性在分子量约为20,000的组分中洗脱。该组分的等电聚焦显示,ThF活性分布在pH值6.5至8.0范围内,而T细胞生长因子(TCGF)活性集中在pH值6.5的单峰处,表明存在一种无TCGF活性的辅助因子。

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