TLR3 activation stimulates cytokine secretion without altering agonist-induced human small airway contraction or relaxation
TLR3 activation stimulates cytokine secretion without altering agonist-induced human small airway contraction or relaxation
复制标题
DOI：
10.1152/ajplung.00133.2009
复制
发表时间：
2009-09-01
影响因子：
4.9
通讯作者：
Panettieri, Reynold A., Jr.
中科院分区：
文献类型：
--
作者：
Cooper, Philip R.;Lamb, Roberta;Panettieri, Reynold A., Jr.
库珀公关，Lamb R，Day ND，Branigan PJ，Kajekar R，San Mateo L，Hornby PJ，Panettieri RA Jr.TLR3的激活刺激细胞因子的分泌，而不改变激动剂诱导的人小气道的收缩或松弛。Am J Physiol肺细胞分子Physiol 297：L530-L537,2009。2009年6月19日首次出版；DOI：10.1152/ajpeng.00133.2009。-呼吸道感染加剧慢性肺部疾病，促进呼吸道炎症和高反应性。Toll样受体3(Toll-like Receptor 3，TLR3)识别病毒双链(Ds)RNA，如多肌苷-多胞苷[Poly(I：C)]，并刺激先天免疫反应。这项研究的目的是验证dsRNA促进肺部炎症和改变人肺切片对胆碱能和β-肾上腺素能受体激动剂的气道反应性的假设。人气道平滑肌(ASM)在Poly(I：C)+/-TNFα和TLR3单抗中孵育24 h。取健康人肺含小气道的精密切割肺切片(PCLS)，分别在0、10、100mU g/mlPoly(I：C)中孵育24 h，用活体显微镜观察肺切片对卡巴胆碱和异丙肾上腺素的收缩和松弛反应。ASM和PCLS培养上清液采用25倍比珠法分析细胞因子分泌情况。在人ASM中，Poly(I：C)(0.5mU g/ml)可增加巨噬细胞炎性蛋白-1α(MIP-1α)和RANTES，而TLR3单抗可阻止这种作用。人PCLS与Poly(I：C)(10和100 mU g/ml)孵育后，对卡巴胆碱和异丙肾上腺素收缩和松弛的对数EC50和最大药物效应(E-max)几乎没有影响。趋化因子和细胞因子的强劲增加证实了相同的人PCLS对Poly(I：C)孵育的反应性。在单独的实验中，PCLS与IL-13或肿瘤坏死因子α(100 ng/ml)孵育可增加对卡巴胆碱的呼吸道敏感性。Poly(I：C)在正常健康人肺切片上促进炎症介质的释放，而这种炎症介质释放与增强的支气管收缩或减弱的支气管扩张无关。由dsRNA启动的TLR3转导刺激下游的先天免疫反应。
Cooper PR, Lamb R, Day ND, Branigan PJ, Kajekar R, San Mateo L, Hornby PJ, Panettieri RA Jr. TLR3 activation stimulates cytokine secretion without altering agonist-induced human small airway contraction or relaxation. Am J Physiol Lung Cell Mol Physiol 297:L530-L537, 2009. First published June 19, 2009; doi:10.1152/ajplung.00133.2009.-Respiratory infections exacerbate chronic lung diseases promoting airway inflammation and hyper-reactivity. Toll-like receptor 3 (TLR3) recognizes viral double-stranded (ds) RNA such as polyinosinic-polycytidylic acid [poly(I:C)] and stimulates innate immune responses. The objective of this study was to test the hypothesis that dsRNA promotes lung inflammation and alters airway responsiveness to cholinergic and beta-adrenergic receptor agonists in human lung slices. Human airway smooth muscle (ASM) was incubated for 24 h in poly(I:C) +/- TNF alpha and a TLR3 monoclonal antibody. Precision-cut lung slices (PCLS; 250-mu m thickness) from healthy human lungs containing a small airway were incubated in 0, 10, or 100 mu g/ml poly(I:C) for 24 h. Intravital microscopy of lung slices was used to quantify contractile and relaxation responsiveness to carbachol and isoproterenol, respectively. Supernatants of ASM and PCLS were analyzed for cytokine secretion using a 25-multiplex bead assay. In human ASM, poly(I:C) (0.5 mu g/ml) increased macrophage inflammatory protein-1 alpha (MIP-1 alpha) and RANTES that was prevented by a TLR3 monoclonal receptor antibody. Incubation of human PCLS with poly(I:C) (10 and 100 mu g/ml) had little effect on the log EC50 or maximum drug effect (E-max) for contraction and relaxation in response to carbachol and isoproterenol, respectively. The responsiveness of the same human PCLS to poly(I:C) incubation was confirmed by the robust increase in chemokines and cytokines. In separate experiments, incubation of PCLS with IL-13 or TNF alpha (100 ng/ml) increased airway sensitivity to carbachol. Poly(I:C) promotes inflammatory mediator release that was not associated with enhanced bronchoconstriction or attenuated bronchodilation in normal healthy human lung slices. Transduction at the TLR3 initiated by dsRNA stimulates downstream innate immune responses.