重症肌无力患者外周血单个核细胞中P2X7受体表达增加与临床严重程度及Th17相关细胞因子血清水平相关。

Increased expression of P2X7 receptor in peripheral blood mononuclear cells correlates with clinical severity and serum levels of Th17-related cytokines in patients with myasthenia gravis.

作者信息

Zhang Yong, Zhang Yanyan, Li Hao, Jia Xiao, Zhang Xiuying, Xia Yan, Wang YuZhong, Fu Linlin, Xiao Chenghua, Geng Deqin

机构信息

Department of Neurology, Affiliated Hospital of Xuzhou Medical university, Xuzhou, Jiangsu, China.

Department of Neurology, Affiliated Hospital of Xuzhou Medical university, Xuzhou, Jiangsu, China; Department of Neurology, The Third People's Hospital of Xuzhou, Xuzhou, Jiangsu, China.

出版信息

Clin Neurol Neurosurg. 2017 Jun;157:88-94. doi: 10.1016/j.clineuro.2017.04.012. Epub 2017 Apr 20.

DOI:10.1016/j.clineuro.2017.04.012

PMID:28458152

Abstract

OBJECTIVES

P2X7R is a well-documented activator of innate and adaptive immune responses. We aimed to measure the expression levels of P2X7R in peripheral blood mononuclear cells (PBMCs) from patients with myasthenia gravis (MG) and to investigate whether the expression of P2X7R is associated with pathogenesis of MG.

PATIENTS AND METHODS

A total of 32 patients with MG (12 generalized MG (GMG) and 20 Ocular MG (OMG) and 22 healthy donors were recruited in this study. The quantitative MG score was used to evaluate the clinical severity. Real-time PCR and western blot were used to measure the levels of P2X7R expressed in PBMCs. Serum Th17-related cytokines (IL-1β, IL-6, IL-17 and IL-21) were tested by ELISA. PBMCs from MG patients were purified and challenged by LPS with or without a selective P2X7R inhibitor (BBG).

RESULTS

Our results showed that the expression of P2X7R mRNA and protein in PBMCs was increased in MG patients compared with healthy controls, with higher expression in generalized patients (GMG) than in ocular patients (OMG). In addition, P2X7R expression presents a significantly positive correlation with clinical severity and serum levels of IL-1β, IL-6, IL-17 and IL-21 in MG. In cultured MG PBMC, LPS challenge led to up-regulated P2X7R expression accompanied with increased production of IL-1β, IL-6, IL-17 and IL-21. Importantly, P2X7R blockade with BBG significantly attenuates the LPS-induced production of cytokines.

CONCLUSION

P2X7R expression was up-regulated in MG and LPS-P2X7R axis may be involved in the pathogenesis of MG by promoting Th17 immune response.

摘要

目的

P2X7R是一种已被充分证明的先天性和适应性免疫反应激活剂。我们旨在检测重症肌无力（MG）患者外周血单个核细胞（PBMC）中P2X7R的表达水平，并研究P2X7R的表达是否与MG的发病机制相关。

患者与方法

本研究共招募了32例MG患者（12例全身型MG（GMG）和20例眼肌型MG（OMG））以及22名健康供者。采用定量MG评分评估临床严重程度。运用实时聚合酶链反应（PCR）和蛋白质免疫印迹法检测PBMC中P2X7R的表达水平。通过酶联免疫吸附测定（ELISA）检测血清中Th17相关细胞因子（白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、白细胞介素-17（IL-17）和白细胞介素-21（IL-21））。对MG患者的PBMC进行纯化，并在有或没有选择性P2X7R抑制剂（BBG）的情况下用脂多糖（LPS）进行刺激。

结果

我们的结果显示，与健康对照相比，MG患者PBMC中P2X7R mRNA和蛋白的表达增加，全身型患者（GMG）的表达高于眼肌型患者（OMG）。此外，MG患者中P2X7R表达与临床严重程度以及IL-1β、IL-6、IL-17和IL-21的血清水平呈显著正相关。在培养的MG PBMC中，LPS刺激导致P2X7R表达上调，同时IL-1β、IL-6、IL-17和IL-21的产生增加。重要的是，用BBG阻断P2X7R可显著减弱LPS诱导的细胞因子产生。