Chaperonin 60.1 (Cpn60.1) is a protein derived from M. tuberculosis that has been shown, along with its peptide fragment IRL201104, to have beneficial effects in models of allergic inflammation. To further investigate the anti-inflammatory properties of Cpn60.1 and IRL201104, we have investigated these molecules in a model of non-allergic lung inflammation. Mice were treated with Cpn60.1 (0.5-5000ng/kg) or IRL201104 (0.00025-2.5ng/kg), immediately before intranasal instillation of bacterial lipopolysaccharide (LPS). Cytokine levels and cell numbers in mouse bronchoalveolar lavage (BAL) fluid were measured 4h after LPS administration. In some experiments mice were depleted of lung-resident phagocytes. Cells from BAL fluid were analysed for inflammasome function. Human umbilical vein endothelial cells (HUVEC) were analysed for adhesion molecule expression. Human neutrophils were analysed for integrin expression, chemotaxis and cell polarisation. Cpn60.1 and IRL201104 significantly inhibited neutrophil migration into the airways, independently of route of administration. This effect of the peptide was absent in TLR4 and Annexin A1 knock-out mice. Intravital microscopy revealed that IRL201104 reduced leukocyte adhesion and migration into inflamed tissues. However, IRL201104 did not significantly affect adhesion molecule expression in HUVEC or integrin expression, chemotaxis or polarisation of human neutrophils at the studied concentrations. In phagocyte-depleted animals, the anti-inflammatory effect of IRL201104 was not significant. IRL201104 significantly reduced IL-1β and NLRP3 expression and increased A20 expression in BAL cells. This study shows that Cpn60.1 and IRL201104 potently inhibit LPS-induced neutrophil infiltration in mouse lungs by a mechanism dependent on tissue-resident phagocytes and to a much lesser extent the pro-resolving factor Annexin A1.

中文翻译：

---

源自伴侣蛋白 60.1 的肽 IRL201104 抑制 LPS 诱导的急性肺部炎症

伴侣蛋白 60.1 (Cpn60.1) 是一种源自结核分枝杆菌的蛋白质，与其肽片段 IRL201104 一起已被证明在过敏性炎症模型中具有有益作用。为了进一步研究 Cpn60.1 和 IRL201104 的抗炎特性，我们在非过敏性肺部炎症模型中研究了这些分子。在鼻内滴注细菌脂多糖 (LPS) 之前，立即用 Cpn60.1 (0.5-5000ng/kg) 或 IRL201104 (0.00025-2.5ng/kg) 处理小鼠。在 LPS 给药后 4 小时测量小鼠支气管肺泡灌洗液 (BAL) 中的细胞因子水平和细胞数量。在一些实验中，小鼠耗尽了肺内的吞噬细胞。分析来自 BAL 液的细胞的炎性体功能。分析人脐静脉内皮细胞 (HUVEC) 的粘附分子表达。分析人中性粒细胞的整联蛋白表达、趋化性和细胞极化。Cpn60.1 和 IRL201104 显着抑制中性粒细胞迁移到气道，与给药途径无关。TLR4 和膜联蛋白 A1 敲除小鼠中不存在肽的这种作用。活体显微镜检查显示 IRL201104 减少了白细胞粘附和迁移到发炎组织中。然而，在研究浓度下，IRL201104 没有显着影响 HUVEC 中粘附分子的表达或整合素的表达、趋化性或人中性粒细胞的极化。在吞噬细胞耗尽的动物中，IRL201104 的抗炎作用不显着。IRL201104 显着降低 BAL 细胞中 IL-1β 和 NLRP3 的表达并增加 A20 的表达。该研究表明，Cpn60.1 和 IRL201104 通过依赖于组织驻留吞噬细胞的机制有效抑制 LPS 诱导的小鼠肺部中性粒细胞浸润，并在较小程度上抑制促解析因子膜联蛋白 A1。