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建立组成性产生细胞毒性细胞生成所需可溶性因子的大鼠-小鼠T细胞杂交瘤:生化与功能特性分析

Establishment of rat-mouse T cell hybridomas that constitutively produce a soluble factor that is needed for the generation of cytotoxic cells: biochemical and functional characterization.

作者信息

Uede T, Kohda H, Ibayashi Y, Kikuchi K

出版信息

J Immunol. 1985 Nov;135(5):3252-7.

PMID:3876381
Abstract

Spleen cells from Lewis rats were cultured with 4 micrograms/ml Con A. These cells were then fused with BW 5147 mouse T lymphoma cells. Two hybrid clones (6B2-B8 and 6B2-E6) obtained by fusion formed CGF effectively. It was found that hybrid cells can be boosted to produce higher levels of CGF upon stimulation with Con A. 6B2-B8 express rat T cell markers. CGF formed by 6B2-B8 had a m.w. of 23,000 and 40,000. CGF was eluted from a Mono Q anion-exchange column with an FPLC system at 0.4 to 0.6 M NaCl as a major peak and at 0.8 M NaCl as a minor peak. CGF was eluted as three peaks with pH 4.1, 4.8, and 5.2 from a Mono P chromatofocusing column. CGF from 6B2-B8 does not contain IL-1, IL-2, IL-3, or CSF.

摘要

将来自Lewis大鼠的脾细胞与4微克/毫升的刀豆蛋白A一起培养。然后将这些细胞与BW 5147小鼠T淋巴瘤细胞融合。通过融合获得的两个杂交克隆(6B2 - B8和6B2 - E6)能有效地形成CGF。发现杂交细胞在用刀豆蛋白A刺激后可被促进产生更高水平的CGF。6B2 - B8表达大鼠T细胞标志物。由6B2 - B8形成的CGF的分子量为23,000和40,000。使用FPLC系统,CGF从Mono Q阴离子交换柱上以0.4至0.6 M NaCl作为主峰和以0.8 M NaCl作为次峰被洗脱下来。CGF从Mono P色谱聚焦柱上以pH 4.1、4.8和5.2被洗脱为三个峰。来自6B2 - B8的CGF不含白细胞介素 - 1、白细胞介素 - 2、白细胞介素 - 3或集落刺激因子。

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