MODULATION OF T CELL ACTIVITY BY 2-ARACHIDONYL GLYCEROL

描述：（申请人提供）： 该实验室以前的研究表明， IL-2的浓度依赖性抑制，通过假定的内源性 大麻素，2-花生四烯酸甘油（2-AG），在活化的T细胞，但它还没有 尚不清楚这种抑制是否通过大麻素受体介导， CB 1和/或CB 2。CB 2受体是特别感兴趣的，因为它具有一个 比CB 1受体的表达水平高得多。我们的大多数 到目前为止，观察是在鼠T细胞中进行的。我们现在感兴趣的 表征2-AG在两种人T细胞系Jurkat E6-1和Jurkat E6 - 2中的作用。 HPB-ALL，其分别表达异常和正常CB 2受体。的 Jurkat和HPB-ALL细胞系与CB 2拮抗剂SR 144528联合， 这将使我们能够确定2-AG在人类T细胞中的活性是否是介导的 CB 2受体。初步研究得出以下结论 观察结果：a）CB 1/CB 2不能逆转2-AG对IL 2的抑制作用 拮抗剂; B）2-AG抑制HPB-ALL细胞中IL-2的转录，但不抑制HPB-ALL细胞中IL-2的转录。 表达异常CB 2受体的Jurkat细胞;和c）矛盾的是， 2-AG还增强T细胞增殖。这些观察导致了 假设：2-花生四烯酸甘油增强CB 1/CB 2中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。该假设将通过以下方式进行检验： 三个具体目标（SA）：SA 1）表征2-AG对 人T细胞中的IL-2转录和分泌; SA 2）对人T细胞的表征 2-AG对人T细胞中T细胞增殖的影响;和SA 3） 2-AG对细胞周期蛋白依赖性激酶作用的表征 抑制剂，p16，p21和p27，以及丝裂原活化激酶，ERK 1和 ERK 2。

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.