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猫白细胞介素-2的生产参数及部分特性

Parameters of production and partial characterization of feline interleukin 2.

作者信息

Bauer R M, Olsen R G

机构信息

Department of Veterinary Pathobiology, College of Veterinary Medicine, Ohio State University, Columbus 43210-1092.

出版信息

Vet Immunol Immunopathol. 1988 Oct;19(3-4):173-83. doi: 10.1016/0165-2427(88)90105-5.

Abstract

The conditions for the production of feline interleukin 2 (IL-2) from peripheral blood leukocytes (PBL) and splenocytes by concanavalin A (Con A) stimulation are described. Feline IL-2 was quantitated by measuring DNA synthesis in the murine IL-2-dependent cell line, CTLL-20. In addition, feline IL-2 was generated for the maintenance of long-term cultures of Con A-stimulated feline PBL and for biochemical characterization. Finally, IL-2 production was evaluated from the PBL of feline leukemia virus (FeLV)-infected cats. Con A at 9.6 micrograms/ml produced a plateau of peak IL-2 activity from 24 to 48 h following stimulation. The tumor promoter, phorbol myristic acetate, stimulated feline IL-2 production and enhanced Con A-stimulated feline IL-2 production. Fetal calf serum (FCS) was not required for IL-2 production; however, FCS at 5% (v/v) allowed for maximal Con A-stimulated IL-2 production. Feline IL-2 generated from Con A-stimulated splenocytes migrated with an apparent molecular size of 13.7 to 23 kD by gel filtration chromatography and supported the proliferation of Con A-activated feline PBL at a final concentration of 0.3 to 0.9 units/ml.

摘要

本文描述了通过刀豆蛋白A(Con A)刺激从外周血白细胞(PBL)和脾细胞中产生猫白细胞介素2(IL-2)的条件。通过测量鼠IL-2依赖细胞系CTLL-2中的DNA合成来定量猫IL-2。此外,还产生了猫IL-2用于维持Con A刺激的猫PBL的长期培养以及进行生化特性分析。最后,评估了感染猫白血病病毒(FeLV)的猫的PBL中IL-2的产生情况。9.6微克/毫升的Con A在刺激后24至48小时产生了IL-2活性峰值的平稳期。肿瘤启动子佛波醇肉豆蔻酸酯刺激猫IL-2的产生并增强Con A刺激的猫IL-2的产生。IL-2的产生不需要胎牛血清(FCS);然而,5%(v/v)的FCS可使Con A刺激的IL-2产生达到最大值。通过凝胶过滤色谱法,从Con A刺激的脾细胞中产生的猫IL-2迁移时的表观分子大小为13.7至23 kD,并以0.3至0.9单位/毫升的终浓度支持Con A激活的猫PBL的增殖。

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