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恒河猴(猕猴)外周血单个核细胞中5-羟色胺转运体的表达

Expression of serotonin transporters by peripheral blood mononuclear cells of rhesus monkeys (Macaca mulatta).

作者信息

Yang G B, Qiu C L, Aye P, Shao Y, Lackner A A

机构信息

State Key Laboratory for Infectious Disease Prevention and Control, National Center for AIDS/STD Control and Prevention, China-CDC, 27 Nanwei Road, Xuanwu District, Beijing 100050, PR China.

出版信息

Cell Immunol. 2007 Aug;248(2):69-76. doi: 10.1016/j.cellimm.2007.09.001. Epub 2007 Nov 5.

Abstract

It has been well established that serotonin (5-hydroxytryptamine, 5-HT) plays a key role in neuro-endocrine-immune networks, mostly through its receptors and/or transporters. Although the presence of 5-HT receptor mRNAs in peripheral blood mononuclear cells (PBMCs) of rhesus monkeys has been reported, there is little information about serotonin transporter (SERT) expression by these cells. To examine SERT expression at the transcription and translation level, one-step RT-PCR, confocal microscopy and flow cytometry were used to detect SERT mRNA and protein expression by rhesus monkey PBMCs. It was found that SERT mRNA could be detected by RT-PCR from all of the rhesus macaque PBMC RNA samples and the nucleotide sequence of the amplicons was identical to the published SERT mRNA sequence. Low level SERT immunoreactivity was also demonstrated on the surface of rhesus PBMCs by confocal microscopy. Almost all lymphocytes and most monocytes were positive for SERT by flow cytometry. In the 2 rhesus macaques examined by multicolor flow cytometry, SERT(bright) cells were more than 84%, 94%, and 96% among CD20+, CD3+, and CD3+CD4+ lymphocytes respectively. These data demonstrate expression of SERT by rhesus macaque PBMCs, and indicate that rhesus macaques would be suitable models to test the in vivo immune regulatory effects of 5-HT or drugs targeting SERT.

摘要

血清素(5-羟色胺,5-HT)在神经-内分泌-免疫网络中发挥关键作用,这一点已得到充分证实,主要是通过其受体和/或转运体来实现。尽管已有报道称恒河猴外周血单核细胞(PBMCs)中存在5-HT受体mRNA,但关于这些细胞中血清素转运体(SERT)表达的信息却很少。为了在转录和翻译水平检测SERT表达,采用一步法RT-PCR、共聚焦显微镜和流式细胞术来检测恒河猴PBMCs中SERT mRNA和蛋白的表达。结果发现,通过RT-PCR可从所有恒河猴PBMC RNA样本中检测到SERT mRNA,扩增子的核苷酸序列与已发表的SERT mRNA序列相同。共聚焦显微镜也显示恒河猴PBMCs表面存在低水平的SERT免疫反应性。流式细胞术检测显示,几乎所有淋巴细胞和大多数单核细胞的SERT呈阳性。在通过多色流式细胞术检测的2只恒河猴中,CD^20^+、CD^3^+和CD^3^+CD^4^+淋巴细胞中SERT(明亮)细胞分别超过84%、94%和96%。这些数据证明了恒河猴PBMCs中SERT的表达,并表明恒河猴将是测试5-HT或靶向SERT的药物体内免疫调节作用的合适模型。

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