Small Donna M, Zani Marie-Louise, Quinn Derek J, Dallet-Choisy Sandrine, Glasgow Arlene M A, O'Kane Cecilia, McAuley Danny F, McNally Paul, Weldon Sinéad, Moreau Thierry, Taggart Clifford C
Centre for Infection and Immunity, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, Northern Ireland, UK.
CEPR, INSERM U1100/EA6305, University of Tours, Tours, France.
Mol Ther. 2015 Jan;23(1):24-31. doi: 10.1038/mt.2014.162. Epub 2014 Sep 5.
Elafin is a serine protease inhibitor produced by epithelial and immune cells with anti-inflammatory properties. Research has shown that dysregulated protease activity may elicit proteolytic cleavage of elafin, thereby impairing the innate immune function of the protein. The aim of this study was to generate variants of elafin (GG- and QQ-elafin) that exhibit increased protease resistance while retaining the biological properties of wild-type (WT) elafin. Similar to WT-elafin, GG- and QQ-elafin variants retained antiprotease activity and susceptibility to transglutaminase-mediated fibronectin cross-linking. However, in contrast to WT-elafin, GG- and QQ-elafin displayed significantly enhanced resistance to degradation when incubated with bronchoalveolar lavage fluid from patients with cystic fibrosis. Intriguingly, both variants, particularly GG-elafin, demonstrated improved lipopolysaccharide (LPS) neutralization properties in vitro. In addition, GG-elafin showed improved anti-inflammatory activity in a mouse model of LPS-induced acute lung inflammation. Inflammatory cell infiltration into the lung was reduced in lungs of mice treated with GG-elafin, predominantly neutrophilic infiltration. A reduction in MCP-1 levels in GG-elafin treated mice compared to the LPS alone treatment group was also demonstrated. GG-elafin showed increased functionality when compared to WT-elafin and may be of future therapeutic relevance in the treatment of lung diseases characterized by a protease burden.
弹性蛋白酶抑制因子是一种由上皮细胞和免疫细胞产生的具有抗炎特性的丝氨酸蛋白酶抑制剂。研究表明，蛋白酶活性失调可能引发弹性蛋白酶抑制因子的蛋白水解切割，从而损害该蛋白的固有免疫功能。本研究的目的是生成弹性蛋白酶抑制因子的变体（GG-弹性蛋白酶抑制因子和QQ-弹性蛋白酶抑制因子），这些变体在保留野生型（WT）弹性蛋白酶抑制因子生物学特性的同时，表现出更高的蛋白酶抗性。与WT-弹性蛋白酶抑制因子类似，GG-和QQ-弹性蛋白酶抑制因子变体保留了抗蛋白酶活性以及对转谷氨酰胺酶介导的纤连蛋白交联的敏感性。然而，与WT-弹性蛋白酶抑制因子不同的是，当与囊性纤维化患者的支气管肺泡灌洗液一起孵育时，GG-和QQ-弹性蛋白酶抑制因子对降解的抗性显著增强。有趣的是，这两种变体，尤其是GG-弹性蛋白酶抑制因子，在体外表现出改善的脂多糖（LPS）中和特性。此外，在LPS诱导的急性肺炎症小鼠模型中，GG-弹性蛋白酶抑制因子显示出增强的抗炎活性。用GG-弹性蛋白酶抑制因子处理的小鼠肺部炎症细胞浸润减少，主要是中性粒细胞浸润。与单独用LPS处理的组相比，在GG-弹性蛋白酶抑制因子处理的小鼠中也证实了MCP-1水平的降低。与WT-弹性蛋白酶抑制因子相比，GG-弹性蛋白酶抑制因子显示出增强的功能，并且在治疗以蛋白酶负荷为特征的肺部疾病方面可能具有未来的治疗相关性。
