MODULATION OF T CELL ACTIVITY BY 2-ARACHIDONYL GLYCEROL

2-花生四烯酰甘油对 T 细胞活性的调节

• 批准号: 6718435
• 负责人: Cheryl Elizabeth Rockwell
• 金额: $ 2.71万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-16 至 2005-03-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6718435
• 关键词: T lymphocyte; cannabinoids; cell growth regulation; clone cells; cyclin dependent kinase; densitometry; enzyme inhibitors; enzyme linked immunosorbent assay; glycerol; human tissue; immunoregulation; immunosuppression; inhibitor /antagonist; interleukin 2; laboratory mouse; leukocyte activation /transformation; mitogen activated protein kinase; polymerase chain reaction; receptor expression; scintillation counter; spectrometry; western blottings

DESCRIPTION: (Provided by Applicant): Previous studies in this laboratory have demonstrated a marked and concentration-dependent inhibition of IL-2 by a putative endogenous cannabinoid, 2-arachidonyl glycerol (2-AG), in activated T cells, but it is yet unknown whether this inhibition is mediated through the cannabinoid receptors, CB1 and/or CB2. The CB2 receptor is of particular interest because it has a much higher level of expression than the CB1 receptor. The majority of our observations have thus far been in murine T cells. We are now interested in characterizing the effect of 2-AG in two human T cell lines, Jurkat E6-1 and HPB-ALL, which express an aberrant and normal CB2 receptor, respectively. The Jurkat and HPB-ALL cell lines in conjunction with the CB2 antagonist, SR144528, will allow us to determine if the activity of 2-AG in human T cells is mediated through the CB2 receptor. Preliminary studies led to the following observations: a) the inhibition of IL2 by 2-AG was not reversed by CB1/CB2 antagonists; b) 2-AG inhibited IL-2 transcription in HPB-ALL cells, but not in Jurkat cells, which express an aberrant CB2 receptor; and c) paradoxically, 2-AG also enhances T cell proliferation. These observations led to the hypothesis: 2-arachidonyl glycerol enhances T cell proliferation in a CB1/CB2 independent mechanism through alterations in cell cycle control and/or ERK MAP kinases, while inhibiting IL-2 production. This hypothesis will be tested using three specific aims (SA): SA1) characterization of the effects of 2-AG upon IL-2 transcription and secretion in human T cells; SA2) characterization on the effects of 2-AG upon T cell proliferation in human T cells; and SA3) characterization of the effects of 2-AG on the cyclin dependent kinase inhibitors, p16, p21, and p27, and the mitogen-activated kinases, ERK1 and ERK2.

描述：（由申请人提供）： 该实验室之前的研究已经证明了显着且 推定的内源性物质对 IL-2 的浓度依赖性抑制 大麻素，2-花生四烯酸甘油（2-AG），存在于活化的 T 细胞中，但尚未 未知这种抑制是否是通过大麻素受体介导的， CB1 和/或 CB2。 CB2 受体特别令人感兴趣，因为它具有 表达水平比 CB1 受体高得多。我们的大多数 迄今为止，观察结果是在小鼠 T 细胞中进行的。我们现在感兴趣的是 表征 2-AG 在两种人类 T 细胞系 Jurkat E6-1 和 HPB-ALL，分别表达异常和正常的 CB2 受体。这 Jurkat 和 HPB-ALL 细胞系与 CB2 拮抗剂 SR144528 联用， 将使我们能够确定人类 T 细胞中 2-AG 的活性是否受到介导 通过CB2受体。初步研究得出以下结论 观察结果：a) 2-AG 对 IL2 的抑制不会被 CB1/CB2 逆转 对手； b) 2-AG 在 HPB-ALL 细胞中抑制 IL-2 转录，但在 Jurkat 细胞，表达异常的 CB2 受体； c) 矛盾的是， 2-AG 还可以增强 T 细胞增殖。这些观察导致 假设：2-花生四烯酸甘油可增强 CB1/CB2 中的 T 细胞增殖 通过改变细胞周期控制和/或 ERK MAP 的独立机制 激酶，同时抑制 IL-2 的产生。该假设将使用以下方法进行检验 三个具体目标 (SA)： SA1) 表征 2-AG 对 人T细胞中IL-2的转录和分泌； SA2) 表征 2-AG 对人 T 细胞增殖的影响；和 SA3) 2-AG 对细胞周期蛋白依赖性激酶影响的表征 抑制剂 p16、p21 和 p27，以及丝裂原激活激酶 ERK1 和 ERK2。