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JURKAT人白血病T细胞系组成性产生白细胞介素2 。

Constitutive interleukin 2 production by the JURKAT human leukemic T cell line.

作者信息

Pawelec G, Borowitz A, Krammer P H, Wernet P

出版信息

Eur J Immunol. 1982 May;12(5):387-92. doi: 10.1002/eji.1830120506.

Abstract

Interleukin 2 (IL2) is a lymphokine produced from phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells and characterized biologically by its ability to maintain the in vitro proliferation of activated T cells. In a search for a convenient alternative source of biologically active human IL2, cells from the five established T cell lines, MOLT4, HSB2, CCRF-CEM, RPMI1301 and JURKAT were cultured at high concentrations for 18-36 h (induction cultures), and their cell-free supernatants thereafter screened on IL2-dependent cultured human and mouse T cells. MOLT4, HSB2, RPMI1301, and CCRF-CEM all failed to produce detectable levels of IL2. Of the three JURKAT cell lines obtained from different sources, one, designated JMN, produced high levels of IL2 activity. A second, JM, failed to produce any IL2, while the third, JHAN, produced intermediate levels. Stimulation of the IL2-producing JMN or JHAN variants with PHA, the phorbol diester 12-0-tetradecanoyl-phorbol-13-acetate (TPA), or both PHA and TPA together, resulted in an apparent increase of IL2 activity in the culture supernatant when assayed by a short-term tritiated thymidine incorporation test. However, both PHA and TPA added directly to the test cells caused substantial thymidine incorporation. Moreover, the nonproducer line JURKAT-JM could not be converted to an IL2 producer by stimulation with PHA, TPA, or both. When JMN supernatants were used to support actual long-term growth and cloning of T cells in limiting dilution, the constitutively produced IL2 was superior to that produced after PHA and/or TPA stimulation. Addition of TPA, but not of PHA, to lectin and TPA-free JMN IL2 resulted in a decreased ability of such supernatants to support clonal T cell growth, suggesting that TPA had a growth-inhibiting effect. These results show that the continuously growing JURKAT-JMN cell line could provide a suitable source of mitogen-free human IL2.

摘要

白细胞介素2(IL2)是一种由植物血凝素(PHA)刺激的外周血单核细胞产生的淋巴因子,其生物学特性是能够维持活化T细胞的体外增殖。为了寻找一种方便的生物活性人IL2替代来源,将五个已建立的T细胞系MOLT4、HSB2、CCRF-CEM、RPMI1301和JURKAT的细胞在高浓度下培养18 - 36小时(诱导培养),然后将其无细胞上清液在依赖IL2培养的人和小鼠T细胞上进行筛选。MOLT4、HSB2、RPMI1301和CCRF-CEM均未产生可检测水平的IL2。从不同来源获得的三个JURKAT细胞系中,一个命名为JMN的细胞系产生高水平的IL2活性。第二个JM细胞系未产生任何IL2,而第三个JHAN细胞系产生中等水平的IL2。用PHA、佛波酯12 - 0 -十四酰佛波醇-13 -乙酸酯(TPA)或PHA和TPA共同刺激产生IL2的JMN或JHAN变体时,通过短期氚标记胸腺嘧啶核苷掺入试验检测,培养上清液中的IL2活性明显增加。然而,直接添加到测试细胞中的PHA和TPA都会导致大量的胸腺嘧啶核苷掺入。此外,非产生细胞系JURKAT-JM不能通过PHA、TPA或两者刺激转化为IL2产生细胞系。当使用JMN上清液在有限稀释条件下支持T细胞的实际长期生长和克隆时,组成性产生的IL2优于PHA和/或TPA刺激后产生的IL2。向不含凝集素和TPA的JMN IL2中添加TPA而非PHA,会导致此类上清液支持克隆T细胞生长的能力下降,这表明TPA具有生长抑制作用。这些结果表明,持续生长的JURKAT-JMN细胞系可以提供无丝裂原的人IL2的合适来源。

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