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恒河猴外周血单个核细胞中多种5-羟色胺(5-HT)受体类型/亚型的mRNA表达

Expression of mRNA for multiple serotonin (5-HT) receptor types/subtypes by the peripheral blood mononuclear cells of rhesus macaques.

作者信息

Yang Gui-Bo, Qiu Chen-Li, Zhao Hui, Liu Qiang, Shao Yiming

机构信息

State Key Laboratory for Infectious Disease Prevention and Control, People's Republic of China.

出版信息

J Neuroimmunol. 2006 Sep;178(1-2):24-9. doi: 10.1016/j.jneuroim.2006.05.016. Epub 2006 Jun 30.

Abstract

To find out whether rhesus macaque peripheral blood mononuclear cells (PBMCs) express mRNA for 5-HT receptors, blood samples from normal healthy rhesus monkeys were used to isolate PBMCs by Ficoll-paque density gradient centrifugation. Total RNA was extracted from MT-2 cells, Hut-78 cells, naive or phytohemagglutinin (PHA) stimulated human and monkey PBMCs. One tube RT-PCR was performed using primers specific for human 5-HT1A, 5-HT1B, 5-HT1E, 5-HT2A, 5-HT2B, 5-HT2C, 5-HT3, 5-HT4, 5-HT6, and 5-HT7 receptors. Amplicons of expected sizes were obtained from human cell lines as well as both human and monkey PBMCs. Both PHA stimulated human and monkey PBMCs express mRNAs for 5-HT1A, 5-HT1B, 5-HT1E, 5-HT2A, 5-HT3, 5-HT4, 5-HT6 receptor types/subtypes. However, mRNAs for 5-HT1B, 5-HT1E and 5-HT2A cannot be confidently detected in some of the PBMC samples without PHA stimulation. 5-HT2B and 5-HT7 receptor mRNA was not detected in most of the samples and 5-HT2C receptor mNRA was not detected at all. FACS analysis revealed that CD3+ lymphocyte increased more than 20% among lymphocytes in the PHA stimulated PBMCs. These data indicate that similar to human PBMC, rhesus macaque PBMC may express multiple types of 5-HT receptors and the expression profile could change after PHA stimulation due to either the changes in cell composition or changes in gene transcription level. This provided a basis for further studies on the neuroimmunomodulatory interactions of 5-HT in rhesus macaques.

摘要

为了确定恒河猴外周血单个核细胞(PBMCs)是否表达5-羟色胺(5-HT)受体的信使核糖核酸(mRNA),我们使用Ficoll-泛影葡胺密度梯度离心法,从正常健康恒河猴的血液样本中分离出PBMCs。从MT-2细胞、Hut-78细胞、未经刺激或经植物血凝素(PHA)刺激的人及猴PBMCs中提取总RNA。使用针对人5-HT1A、5-HT1B、5-HT1E、5-HT2A、5-HT2B、5-HT2C、5-HT3、5-HT4、5-HT6和5-HT7受体的特异性引物进行单管逆转录聚合酶链反应(RT-PCR)。在人细胞系以及人及猴PBMCs中均获得了预期大小的扩增子。PHA刺激的人及猴PBMCs均表达5-HT1A、5-HT1B、5-HT1E、5-HT2A、5-HT3、5-HT4、5-HT6受体类型/亚型的mRNA。然而,在一些未经PHA刺激的PBMC样本中,无法可靠地检测到5-HT1B、5-HT1E和5-HT2A的mRNA。在大多数样本中未检测到5-HT2B和5-HT7受体mRNA,且完全未检测到5-HT2C受体mRNA。荧光激活细胞分选(FACS)分析显示,在PHA刺激的PBMCs中,淋巴细胞中的CD3 + 淋巴细胞增加了20%以上。这些数据表明,与人类PBMCs相似,恒河猴PBMCs可能表达多种类型的5-HT受体,并且由于细胞组成的变化或基因转录水平的变化,PHA刺激后表达谱可能会发生变化。这为进一步研究恒河猴中5-HT的神经免疫调节相互作用提供了基础。

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