病毒刺激物通过慢性阻塞性肺疾病上皮细胞引发过度的胸腺基质淋巴细胞生成素表达：内体 TLR3 和胞质 RIG-I 样螺旋酶的作用。

Viral stimuli trigger exaggerated thymic stromal lymphopoietin expression by chronic obstructive pulmonary disease epithelium: role of endosomal TLR3 and cytosolic RIG-I-like helicases.

机构信息

Unit of Respiratory Immunopharmacology, Lund University Sweden, Lund, Sweden.

出版信息

J Innate Immun. 2012;4(1):86-99. doi: 10.1159/000329131. Epub 2011 Jun 20.

DOI:10.1159/000329131

PMID:21691053

Abstract

BACKGROUND

Rhinovirus (RV)-induced chronic obstructive pulmonary disease (COPD) exacerbations exhibit TH(2)-like inflammation. We hypothesized that RV-infected bronchial epithelial cells (BEC) overproduce TH(2)-switching hub cytokine, thymic stromal lymphopoietin (TSLP) in COPD.

METHODS

Primary BEC from healthy (HBEC) and from COPD donors (COPD-BEC) were grown in 12-well plates, infected with RV16 (0.5-5 MOI) or stimulated with agonists for either toll-like receptor (TLR) 3 (dsRNA, 0.1-10 μg/ml) or RIG-I-like helicases (dsRNA-LyoVec, 0.1-10 μg/ml). Cytokine mRNA and protein were determined (RTqPCR; ELISA).

RESULTS

dsRNA dose-dependently evoked cytokine gene overproduction of TSLP, CXCL8 and TNF-α in COPD-BEC compared to HBEC. This was confirmed using RV16 infection. IFN-β induction did not differ between COPD-BEC and HBEC. Endosomal TLR3 inhibition by chloroquine dose-dependently inhibited dsRNA-induced TSLP generation and reduced generation of CXCL8, TNF-α, and IFN-β. Stimulation of cytosolic viral sensors (RIG-I-like helicases) with dsRNA-LyoVec increased production of CXCL8, TNF-α, and IFN-β, but not TSLP.

CONCLUSIONS

Endosomal TLR3-stimulation, by dsRNA or RV16, induces overproduction of TSLP in COPD-BEC. dsRNA- and RV-induced overproduction of TNF-α and CXCL8 involves endosomal TLR3 and cytosolic RIG-I-like helicases and so does the generation of IFN-β in COPD-BEC. RV16 and dsRNA-induced epithelial TSLP may contribute to pathogenic effects at exacerbations and development of COPD.

摘要

背景

鼻病毒（RV）引起的慢性阻塞性肺疾病（COPD）加重表现出 TH（2）样炎症。我们假设 RV 感染的支气管上皮细胞（BEC）在 COPD 中过度产生 TH（2）转换枢纽细胞因子，胸腺基质淋巴细胞生成素（TSLP）。

方法

将来自健康人（HBEC）和 COPD 供体（COPD-BEC）的原代 BEC 在 12 孔板中生长，用 RV16（0.5-5MOI）感染或用 TLR（TLR）3（dsRNA，0.1-10μg/ml）或 RIG-I 样螺旋酶的激动剂刺激（dsRNA-LyoVec，0.1-10μg/ml）。测定细胞因子 mRNA 和蛋白（RTqPCR；ELISA）。

结果

与 HBEC 相比，dsRNA 剂量依赖性地引起 COPD-BEC 中 TSLP、CXCL8 和 TNF-α 的细胞因子基因过度产生。这一点通过 RV16 感染得到了证实。COPD-BEC 和 HBEC 之间 IFN-β 的诱导没有差异。氯喹对内体 TLR3 的抑制作用呈剂量依赖性地抑制 dsRNA 诱导的 TSLP 产生，并减少 CXCL8、TNF-α 和 IFN-β的产生。dsRNA-LyoVec 刺激细胞质病毒传感器（RIG-I 样螺旋酶）增加了 CXCL8、TNF-α 和 IFN-β的产生，但不增加 TSLP。

结论

dsRNA 或 RV16 刺激内体 TLR3 可诱导 COPD-BEC 中 TSLP 的过度产生。dsRNA 和 RV 诱导的 TNF-α和 CXCL8 的过度产生涉及内体 TLR3 和细胞质 RIG-I 样螺旋酶，COPD-BEC 中 IFN-β 的产生也是如此。RV16 和 dsRNA 诱导的上皮 TSLP 可能有助于加重和 COPD 的发展中的致病作用。