Unit of Respiratory Immunopharmacology, Lund University Sweden, Lund, Sweden.
J Innate Immun. 2012;4(1):86-99. doi: 10.1159/000329131. Epub 2011 Jun 20.
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.
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).
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.
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.
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）。
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）。
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。
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。
