Identification of a functionally relevant cannabinoid receptor on mouse spleen cells that is involved in cannabinoid-mediated immune modulation.
Identification of a functionally relevant cannabinoid receptor on mouse spleen cells that is involved in cannabinoid-mediated immune modulation.
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DOI：
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发表时间：
1992-11
影响因子：
3.6
通讯作者：
N. Kaminski;M. Abood;F. Kessler;B. Martin;A. R. Schatz
中科院分区：
文献类型：
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作者：
N. Kaminski;M. Abood;F. Kessler;B. Martin;A. R. Schatz
大麻素介导的对中枢神经系统的作用的广泛的行为和生物化学表征已经揭示了至少三条证据，支持推定的鸟嘌呤核苷酸结合蛋白偶联大麻素受体对大麻模拟作用的作用，（i）立体选择性，（ii）腺苷酸环化酶/cAMP第二信使系统的抑制，和（iii）用合成大麻素[3 H]CP-55，940进行的放射性配体结合研究，表明与脑组织制剂的高度特异性结合。基于我们实验室最近的发现，证明δ 9-四氢大麻酚显著抑制毛喉素刺激的小鼠脾细胞中cAMP积累，研究了与小鼠脾细胞相关的鸟嘌呤核苷酸结合蛋白偶联大麻素受体的存在及其在免疫调节中的功能作用。在本研究中，用合成双环大麻素（-）-CP-55，940相对于（+）CP-56，667以及用11-OH-δ 8-四氢大麻素-二甲基庚基（-）-HU-210相对于（+）-HU-211观察到立体选择性免疫调节。在这两种情况下，（-）-对映异构体表现出比（+）-异构体更大的免疫抑制效力，如通过体外绵羊红细胞抗体形成细胞反应所测量的。放射性配体结合研究产生了饱和等温线，表现出约45-65%的特异性结合小鼠脾细胞。Scatchard分析表明脾细胞上有一个单一的结合位点，Kd为910 pM，Bmax约为1000个受体/脾细胞。使用大麻素受体的特异性引物的分离的脾RNA的RNA聚合酶链反应导致与大麻素受体cDNA杂交的854-脱氢酶预测产物的扩增，证明大麻素受体mRNA在小鼠脾中的存在。总之，这些发现强烈支持大麻素受体在大麻模拟剂的免疫调节中的作用。
Extensive behavioral and biochemical characterization of cannabinoid-mediated effects on the central nervous system has revealed at least three lines of evidence supporting the role of a putative guanine nucleotide-binding protein-coupled cannabinoid receptor for cannabimimetic effects, (i) stereoselectivity, (ii) inhibition of the adenylate cyclase/cAMP second messenger system, and (iii) radioligand-binding studies with the synthetic cannabinoid [3H]CP-55,940 indicating a high degree of specific binding to brain tissue preparations. Based on recent findings from our laboratory demonstrating that delta 9-tetrahydrocannabinol markedly inhibited forskolin-stimulated cAMP accumulation in mouse spleen cells, the presence of a guanine nucleotide-binding protein-coupled cannabinoid receptor associated with mouse spleen cells and its functional role in immune modulation were investigated. In the present studies, stereoselective immune modulation was observed with the synthetic bicyclic cannabinoid (-)-CP-55,940 versus (+) CP-56,667 and with 11-OH-delta 8-tetrahydrocannabinol-dimethylheptyl, (-)-HU-210 versus (+)-HU-211. In both cases, the (-)-enantiomer demonstrated greater immunoinhibitory potency than the (+)-isomer, as measured by the in vitro sheep red blood cell antibody-forming cell response. Radioligand binding studies produced a saturation isotherm exhibiting approximately 45-65% specific binding to mouse spleen cells. Scatchard analysis demonstrated a single binding site on spleen cells, possessing a Kd of 910 pM and a Bmax of approximately 1000 receptors/spleen cell. RNA polymerase chain reaction of isolated splenic RNA using specific primers for the cannabinoid receptor resulted in the amplification of a 854-kilobase predicted product that hybridized with cannabinoid receptor cDNA, demonstrating the presence of cannabinoid receptor mRNA in mouse spleen. Together, these findings strongly support the role of a cannabinoid receptor in immune modulation by cannabimimetic agents.