Cannabinoid receptors CB1 and CB2: A characterization of expression and adenylate cyclase modulation within the immune system
Cannabinoid receptors CB1 and CB2: A characterization of expression and adenylate cyclase modulation within the immune system
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DOI：
10.1006/taap.1996.8034
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发表时间：
1997-02-01
影响因子：
3.8
通讯作者：
Kaminski, NE
中科院分区：
文献类型：
--
作者：
Schatz, AR;Lee, M;Kaminski, NE
大麻素受体（CB）表达在免疫细胞和组织制剂中进行了表征。 Northern 分析显示，小鼠脾脏、大脑以及大鼠小脑中的 CB1（脑型）转录物与 6 kb 相似。 Northern 分析未在小鼠胸腺或大鼠脾脏 RNA 中检测到 CB1。检测到 CB2（外周）在小鼠脾脏和胸腺中与 4-kb 转录物相似，在大鼠脾脏中与 2.4-kb 转录物相似。小鼠脾脏和胸腺中 CB2 转录物的定量显示，分别类似于 4 x 10(3) 和 4 x 10(2) 分子/100 ng RNA，而小鼠大脑中没有可量化的 CB2。相反，CB1 在小鼠脑中表达（类似于 2 x 10(5) 分子/100 ng RNA），在小鼠脾脏中表达较低（类似于 2 x 10(2) 分子/100 ng RNA），并且在小鼠胸腺中无法定量。完整小鼠脾细胞中的竞争结合表明，非放射性标记的大麻素 CP-55940、Win-55212-2、CP-56667、Delta(9)-THC 和大麻酚均与 H-3-CP-55940（一种高亲和力非歧视性 CB1 和 CB2 受体配体）竞争受体结合。先前的研究结果表明大麻素可显着抑制 T 细胞依赖性免疫反应，因此对原代 T 细胞和几种 T 细胞系进行了表征。放射性配体结合分析在纯化的脾 T 细胞中鉴定出 100-300 个大麻素受体结合位点/细胞，K-d 约为 200-700 pM，该细胞也表现出大麻素诱导的腺苷酸环化酶抑制作用。人 T 细胞系的 Northern 分析显示与 2.4-kb CB2 mRNA 转录物相似，但 HPB-ALL 细胞中没有 CB1，该细胞系也表现出腺苷酸抑制作用Delta(9)-THC 环化酶。相反，Jurkat E6-1 细胞表达了一种不寻常的 CB2 mRNA 带型模式，表达了三种不同的转录本大小，其中没有一个是 2.4 kb（人 CB2 的大小）。 Jurkat 也不表达 CB1 mRNA，并且在用 Delta(9)-THC 处理时没有表现出对腺苷酸环化酶的抑制。总的来说，这些结果进一步证明 CB2 是免疫系统内主要的大麻素受体，并且这种形式的受体在 T 细胞上表达。 (C) 1997 年学术出版社。
Cannabinoid receptor (CB) expression was characterized in immunological cell and tissue preparations. Northern analysis revealed similar to 6-kb transcripts for CB1 (brain-type) in mouse spleen and brain and in rat cerebellum. CB1 was not detected in mouse thymus or rat spleen RNA by Northern analysis. CB2 (peripheral) was detected as a similar to 4-kb transcript in mouse spleen and thymus and as similar to 2.4-kb transcripts in rat spleen. Quantitation of CB2 transcripts in mouse spleen and thymus revealed similar to 4 x 10(3) and similar to 4 x 10(2) molecules/100 ng RNA, respectively, with no quantifiable CB2 in mouse brain. Conversely, CB1 was expressed in mouse brain (similar to 2 x 10(5) molecules/100 ng RNA) with lower expression in mouse spleen (similar to 2 x 10(2) molecules/100 ng RNA) and was not quantifiable in mouse thymus. Competition binding in intact mouse splenocytes demonstrated that nonradiolabeled cannabinoids CP-55940, Win-55212-2, CP-56667, Delta(9)-THC, and cannabinol all competed for receptor binding with H-3-CP-55940, a high-affinity nondiscriminating CB1 and CB2 receptor ligand. Based on previous findings which demonstrated a marked inhibition of T-cell-dependent immune responses by cannabinoids, primary T cells and several T-cell lines were characterized. Radioligand binding analysis identified 100-300 cannabinoid receptor binding sites/cell with an approximate K-d of 200-700 pM in purified splenic T cells which also exhibited cannabinoid-induced inhibition of adenylate cyclase, Northern analysis of human T-cell lines revealed similar to 2.4-kb CB2 mRNA transcripts but no CB1 in HPB-ALL cells, a cell line which also exhibited inhibition of adenylate cyclase by Delta(9)-THC. Conversely, Jurkat E6-1 cells expressed an unusual mRNA banding pattern for CB2 expressing three distinct transcript sizes, none of which were 2.4 kb, the size for human CB2. Jurkat also did not express CB1 mRNA and did not exhibit inhibition of adenylate cyclase when treated with Delta(9)-THC. Collectively, these results provide further evidence that CB2 is the predominant cannabinoid receptor within the immune system and that this form of the receptor is expressed on T cells. (C) 1997 Academic Press.