Phialophora verrucosa (P. verrucosa) is a pathogen that can cause chromoblastomycosis and phaeohyphomycosis. Recent evidence suggests that neutrophils can produce neutrophil extracellular traps (NETs) that can protect against invasive pathogens. As such, we herein explored the in vitro functional importance of P. verrucosa-induced NET formation. By assessing the co-localization of neutrophil elastase and DNA, we were able to confirm the formation of classical NETs entrapping P. verrucosa specimens. Sytox Green was then used to stain these NETs following neutrophil infection with P. verrucosa in order to quantify the formation of these extracellular structures. NET formation was induced upon neutrophil exposure to both live, UV-inactivated, and dead P. verrucosa fungi. The ability of these NETs to kill fungal hyphae and conidia was demonstrated through MTT and pouring plate assays, respectively. Overall, our results confirmed that P. verrucosa was able to trigger the production of NETs, suggesting that these extracellular structures may represent an important innate immune effector mechanism controlling physiological responses to P. verrucosa infection, thereby aiding in pathogen control during the acute phases of infection.

Phialophora verrucosa（P. verrucosa）是一种病原体，可引起色素芽生菌病和褐丝菌病。最近的证据表明，中性粒细胞可以产生中性粒细胞胞外陷阱 (NETs)，可以防止侵入性病原体。因此，我们在此探讨了疣状假单胞菌诱导的 NET 形成的体外功能重要性。通过评估中性粒细胞弹性蛋白酶和 DNA 的共定位，我们能够确认经典 NET 的形成捕获了P. verrucosa标本。在中性粒细胞感染P. verrucosa后，使用 Sytox Green 对这些 NET 进行染色为了量化这些细胞外结构的形成。NET 形成是在中性粒细胞暴露于活的、紫外线灭活的和死的P. verrucosa真菌时诱导的。这些 NETs 杀死真菌菌丝和分生孢子的能力分别通过 MTT 和浇注板测定证明。总的来说，我们的结果证实了疣状假单胞菌能够触发 NET 的产生，这表明这些细胞外结构可能代表了一种重要的先天免疫效应机制，控制了对疣状假单胞菌感染的生理反应，从而有助于在急性期控制病原体。感染。