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干燥综合征抗原B作为内源性危险分子诱导多形核中性粒细胞中白细胞介素-8基因表达。

Sjögren's Syndrome Antigen B Acts as an Endogenous Danger Molecule to Induce Interleukin-8 Gene Expression in Polymorphonuclear Neutrophils.

作者信息

Wu Cheng-Han, Li Ko-Jen, Yu Chia-Li, Tsai Chang-Youh, Hsieh Song-Chou

机构信息

Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine, Taipei, Taiwan; Division of Immunology, Rheumatology and Allergy, Department of Internal Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.

Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; Division of Immunology, Rheumatology and Allergy, Department of Internal Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.

出版信息

PLoS One. 2015 Apr 27;10(4):e0125501. doi: 10.1371/journal.pone.0125501. eCollection 2015.

DOI:10.1371/journal.pone.0125501
PMID:25915936
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4411107/
Abstract

BACKGROUND

Sjögren's syndrome antigen B is expressed in the nucleus and surface membrane of human polymorphonuclear neutrophils and is released after cell death. However, its biological role is not clear. This study is aimed to investigate the effect of Sjögren's syndrome antigen B on human polymorphonuclear neutrophils.

METHODS

Human recombinant Sjögren's syndrome antigen B (rSSB) purified from E. coli was incubated with human polymorphonuclear neutrophils as well as retinoid acid-induced granulocytic differentiated HL-60 cells, HL-60 (RA). Interleukin (IL)-8 protein production and mRNA expressions were measured by enzyme-linked immunosorbent assay and quantitative-polymerase chain reaction, respectively. Uptake of fluorescein isothiocyanate (FITC)-rSSB was assessed by flow cytometry and fluorescence microscopy. Moreover, mitogen-activated protein kinase (MAPK) pathways and nuclear factor-kappaB activation were investigated.

RESULTS

Human rSSB stimulated IL-8 production from normal human neutrophils and HL-60 (RA) cells in a time- and dose-dependent manner. This IL-8-stimulated activity was blocked by chloroquine and NH4Cl, indicating that endosomal acidification is important for this effect. We found rSSB activated both MAPK pathway and nuclear factor-kappaB signaling to transcribe the IL-8 gene expression of cells. Furthermore, tumor necrosis factor-α exerted an additive effect and rSSB-anti-SSB immune complex exhibited a synergistic effect on rSSB-induced IL-8 production.

CONCLUSIONS

Sjögren's syndrome antigen B might act as an endogenous danger molecule to enhance IL-8 gene expression in human polymorphonuclear neutrophils.

摘要

背景

干燥综合征抗原B在人多形核中性粒细胞的细胞核和表面膜中表达，并在细胞死亡后释放。然而，其生物学作用尚不清楚。本研究旨在探讨干燥综合征抗原B对人多形核中性粒细胞的影响。

方法

将从大肠杆菌中纯化的人重组干燥综合征抗原B（rSSB）与人多形核中性粒细胞以及视黄酸诱导的粒细胞分化的HL-60细胞HL-60（RA）一起孵育。分别通过酶联免疫吸附测定和定量聚合酶链反应测量白细胞介素（IL）-8蛋白的产生和mRNA表达。通过流式细胞术和荧光显微镜评估异硫氰酸荧光素（FITC）-rSSB的摄取。此外，还研究了丝裂原活化蛋白激酶（MAPK）途径和核因子-κB的激活。

结果

人rSSB以时间和剂量依赖性方式刺激正常人中性粒细胞和HL-60（RA）细胞产生IL-8。氯喹和NH4Cl可阻断这种IL-8刺激活性，表明内体酸化对该效应很重要。我们发现rSSB激活了MAPK途径和核因子-κB信号传导，从而转录细胞的IL-8基因表达。此外，肿瘤坏死因子-α发挥了相加作用，rSSB-抗SSB免疫复合物对rSSB诱导的IL-8产生表现出协同作用。

结论

干燥综合征抗原B可能作为一种内源性危险分子，增强人多形核中性粒细胞中IL-8基因的表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/a166b9a9193e/pone.0125501.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/698885e87a2a/pone.0125501.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/cc1f115c836f/pone.0125501.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/3a411e90775b/pone.0125501.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/e2ddd54c2c0a/pone.0125501.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/b7c8c73b11fc/pone.0125501.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/a166b9a9193e/pone.0125501.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/698885e87a2a/pone.0125501.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/cc1f115c836f/pone.0125501.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/3a411e90775b/pone.0125501.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/e2ddd54c2c0a/pone.0125501.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/b7c8c73b11fc/pone.0125501.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/109f/4411107/a166b9a9193e/pone.0125501.g006.jpg

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