慢性丙型肝炎患者 PBMCs 中 P2X 受体的转录分析：抗病毒治疗反应和 HCV 诱导发病机制的新见解。

Transcript analysis of P2X receptors in PBMCs of chronic HCV patients: an insight into antiviral treatment response and HCV-induced pathogenesis.

机构信息

1 Atta ur Rahman School of Applied Bio-Sciences, National University of Sciences & Technology, Islamabad.

出版信息

Viral Immunol. 2013 Oct;26(5):343-50. doi: 10.1089/vim.2013.0044.

DOI:10.1089/vim.2013.0044

PMID:24116708

Abstract

BACKGROUND

After invasion of hepatocytes and immune cells, hepatitis C virus has the ability to escape from the host immune system, leading to the progression of disease into chronic infection with associated liver morbidities. Adenosine 5'triphosphate (ATP) is released in most of the pathological events from the affected cells and acts as a signaling molecule by binding to P2X receptors expressed on the host's immune cells and activates the immune system for pro-inflammatory response. Therefore, the present study was designed to analyze the transcript expression of the ionotropic purinergic P2X receptors on peripheral blood mononuclear cells (PBMCs) of chronic HCV patients to have study the immune responses mediated by P2X receptors in chronic HCV infections.

METHODS

PBMCs were isolated from the collected blood samples. Transcript analysis of P2X receptors in PBMCs was done. The identity of amplified product was confirmed by sequencing PCR, while the quantification of the transcript expression was done by real time PCR. The relative expression of the P2X receptors was analyzed by unpaired Student's t test using GraphPad Prims 5 software.

RESULTS

We found that out of seven isoforms of P2X receptors, P2X1, P2X4, P2X5, and P2X7 receptors are expressed on the PBMCs. P2X1 and P2X7 are significantly upregulated in treatment-naïve chronic HCV patients by 2.2- and 2.5-fold, respectively. However, only P2X7 expression is found increased by 2.7-fold in patients achieving sustained virological response (SVR) after antiviral treatment compared to healthy controls. The expression of P2X receptors remained unaltered in chronic HCV patients not responding to the treatment.

CONCLUSION

The present study confirms the significant involvement of P2X receptors in the immune responses mediated by the PBMCs in the chronic HCV infection, which should be further investigated to devise strategies to augment the immune system against this chronic viral disease.

摘要

背景

丙型肝炎病毒（HCV）侵入肝细胞和免疫细胞后，具有逃避宿主免疫系统的能力，导致疾病进展为慢性感染，并伴有肝脏病变。三磷酸腺苷（ATP）在受影响细胞的大多数病理事件中释放，并通过与宿主免疫细胞上表达的 P2X 受体结合作为信号分子发挥作用，激活免疫系统引起促炎反应。因此，本研究旨在分析慢性 HCV 患者外周血单个核细胞（PBMC）中离子型嘌呤能 P2X 受体的转录表达，以研究 P2X 受体在慢性 HCV 感染中的免疫反应。

方法

从采集的血液样本中分离 PBMC。对 PBMC 中的 P2X 受体进行转录分析。通过测序 PCR 确认扩增产物的身份，通过实时 PCR 定量转录表达。使用 GraphPad Prims 5 软件通过未配对学生 t 检验分析 P2X 受体的相对表达。

结果

我们发现，在七种 P2X 受体亚型中，P2X1、P2X4、P2X5 和 P2X7 受体在 PBMC 上表达。未接受治疗的慢性 HCV 患者中，P2X1 和 P2X7 的表达分别上调了 2.2 倍和 2.5 倍。然而，仅在抗病毒治疗后获得持续病毒学应答（SVR）的患者中，P2X7 的表达增加了 2.7 倍，与健康对照组相比。未对治疗产生反应的慢性 HCV 患者的 P2X 受体表达保持不变。