Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden.
Department of Immunology, Genetics and Pathology - BioVis, Uppsala University, Uppsala, Sweden.
J Innate Immun. 2022;14(5):433-446. doi: 10.1159/000520972. Epub 2021 Dec 22.
Previous research has indicated an intimate functional communication between mast cells (MCs) and neutrophils during inflammatory conditions, but the nature of such communication is not fully understood. Activated neutrophils are known to release DNA-containing extracellular traps (neutrophil extracellular traps [NETs]) and, based on the known ability of tryptase to interact with negatively charged polymers, we here hypothesized that tryptase might interact with NET-contained DNA and thereby regulate NET formation. In support of this, we showed that tryptase markedly enhances NET formation in phorbol myristate acetate-activated human neutrophils. Moreover, tryptase was found to bind vividly to the NETs, to cause proteolysis of core histones and to cause a reduction in the levels of citrullinated histone-3. Secretome analysis revealed that tryptase caused increased release of numerous neutrophil granule compounds, including gelatinase, lactoferrin, and myeloperoxidase. We also show that DNA can induce the tetrameric, active organization of tryptase, suggesting that NET-contained DNA can maintain tryptase activity in the extracellular milieu. In line with such a scenario, DNA-stabilized tryptase was shown to efficiently degrade numerous pro-inflammatory compounds. Finally, we showed that tryptase is associated with NET formation in vivo in a melanoma setting and that NET formation in vivo is attenuated in mice lacking tryptase expression. Altogether, these findings reveal that NET formation can be regulated by MC tryptase, thus introducing a novel mechanism of communication between MCs and neutrophils.
先前的研究表明,在炎症条件下,肥大细胞(MCs)和中性粒细胞之间存在密切的功能通讯,但这种通讯的性质尚不完全清楚。已知激活的中性粒细胞会释放含有 DNA 的细胞外陷阱(中性粒细胞细胞外陷阱[NETs]),并且基于已知的类胰蛋白酶与带负电荷的聚合物相互作用的能力,我们假设类胰蛋白酶可能与 NET 中包含的 DNA 相互作用,从而调节 NET 的形成。支持这一观点的是,我们表明类胰蛋白酶显着增强了佛波醇肉豆蔻酸酯激活的人中性粒细胞中的 NET 形成。此外,发现类胰蛋白酶强烈结合 NET,导致核心组蛋白的蛋白水解,并导致瓜氨酸化组蛋白-3 的水平降低。分泌组分析显示,类胰蛋白酶导致许多中性粒细胞颗粒化合物的释放增加,包括明胶酶、乳铁蛋白和髓过氧化物酶。我们还表明 DNA 可以诱导类胰蛋白酶的四聚体、活性组织,这表明 NET 中包含的 DNA 可以在细胞外环境中维持类胰蛋白酶的活性。与此情况一致,DNA 稳定的类胰蛋白酶有效地降解了许多促炎化合物。最后,我们表明在黑色素瘤环境中,类胰蛋白酶与 NET 的形成有关,并且缺乏类胰蛋白酶表达的小鼠体内的 NET 形成受到抑制。总而言之,这些发现揭示了 NET 的形成可以通过 MC 类胰蛋白酶来调节,从而为 MCs 和中性粒细胞之间的通讯引入了一种新的机制。