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诱导T细胞杂交瘤细胞溶解活性及T细胞生长因子受体表达的可溶性因子的特性分析

Characterization of soluble factors that induce the cytolytic activity and the expression of T cell growth factor receptors of a T cell hybrid.

作者信息

Erard F, Corthesy P, Smith K A, Fiers W, Conzelmann A, Nabholz M

出版信息

J Exp Med. 1984 Aug 1;160(2):584-99. doi: 10.1084/jem.160.2.584.

Abstract

A rat X mouse T cell hybrid (PC60) proliferates in the absence of T cell growth factor (TCGF) and its cytolytic activity can be induced by culture in mixed leukocyte culture supernatants or concanavalin A-activated rat spleen cell supernatant (CS) to lyse 51Cr-labeled tumor target cells. To characterize the factor(s) responsible for this reversible induction, serum-free CS was fractionated by reverse phase high performance liquid chromatography and by phenyl-Sepharose chromatography. A cytotoxicity-inducing activity (CIA) was separated from TCGF and macrophage-activating factor/interferon-gamma. CIA was found to be a macromolecule with an apparent molecular weight of 12,000-18,000 and a pI of 5.0 and 6.2. Its activity on PC60 cells depended on the addition of TCGF. Thus TCGF may have other effects on T cells than the induction of entry into cell cycle. The number of TCGF surface receptors on PC60 cells was measured using purified 3H-TCGF. TCGF receptors were undetectable on noninduced cells but appeared during induction. The expression of TCGF receptors was not induced either by TCGF or by CIA-containing supernatants or fractions alone, only by a combination of both. These results show that TCGF plays a role in the regulation of the expression of its own receptors.

摘要

一种大鼠X小鼠T细胞杂交体(PC60)在没有T细胞生长因子(TCGF)的情况下也能增殖,并且其细胞溶解活性可通过在混合白细胞培养上清液或伴刀豆球蛋白A激活的大鼠脾细胞上清液(CS)中培养来诱导,从而裂解51Cr标记的肿瘤靶细胞。为了鉴定负责这种可逆诱导的因子,通过反相高效液相色谱法和苯基-琼脂糖色谱法对无血清CS进行了分级分离。细胞毒性诱导活性(CIA)与TCGF以及巨噬细胞激活因子/干扰素-γ分离开来。发现CIA是一种大分子,其表观分子量为12,000 - 18,000,pI为5.0和6.2。它对PC60细胞的活性依赖于TCGF的添加。因此,TCGF对T细胞可能具有除诱导进入细胞周期之外的其他作用。使用纯化的3H-TCGF测量PC60细胞上TCGF表面受体的数量。在未诱导的细胞上检测不到TCGF受体,但在诱导过程中出现。TCGF受体的表达既不是由TCGF单独诱导,也不是由含CIA的上清液或级分单独诱导,而是仅由两者的组合诱导。这些结果表明,TCGF在其自身受体表达的调节中发挥作用。

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