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

被引:29
作者
Yang, Gui-Bo [1 ]
Qiu, Chen-Li [1 ]
Zhao, Hui [1 ]
Liu, Qiang [1 ]
Shao, Yiming [1 ]
机构
[1] China CDC, Natl Ctr AIDS STD Control & Prevent, State Key Lab Infect Dis Prevent & Control, Beijing 100050, Peoples R China
基金
中国国家自然科学基金;
关键词
serotonin; 5-HT receptors; one-step RT-PCR; PBMCs; Macaca mulatta;
D O I
10.1016/j.jneuroim.2006.05.016
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
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-HT1(A), 5-HT1(B), 5-HT1(E), 5-HT2(A), 5-HT2(B), 5-HT2(C), 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-HT1(A), 5-HT1(B), 5-HT1(E), 5-HT2(A), 5-HT3, 5-HT4, 5-HT6 receptor types/subtypes. However, mRNAs for 5-HT1(B), 5-HT1(E) and 5-HT2(A) cannot be confidently detected in some of the PBMC samples without PHA stimulation. 5-HT2(B) and 5-HT7 receptor mRNA was not detected in most of the samples and 5-HT2(C) receptor mNRA was not detected at all. FACS analysis revealed that CD3+ lymphocyte increased more than 20% among lymphocytes in the PffA 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. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:24 / 29
页数:6
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