■中性粒细胞是已知的先天免疫介质,然而,它们在疱疹病毒感染中的效应子功能仍然知之甚少。这里,我们阐明了中性粒细胞胞外诱捕网(NETs)在单纯疱疹病毒1型(HSV-1)眼部感染中的机制作用和关键作用.
■从HSV-1感染的小鼠中收集中性粒细胞,荧光成像,和免疫印迹分析。在L.V.Prasad眼科研究所收集健康受试者和HSV-1患者和小鼠的泪液样本,印度,在伊利诺伊大学,美国,分别。对于体内研究,用HSV-1(McKrae株)感染C57BL/6小鼠以及多样性近交小鼠,然后在不同时间点(0-10天)收集泪液。样品用于流式细胞术,ELISA,和免疫荧光分析。使用角膜炎数据集的人类转录组谱评估NETosis信号通路。我们还进行了中性粒细胞耗竭研究。
■我们的数据显示,主要在受感染的眼睛中形成可辨别的时间网(NETosis),在正常和多样性近交小鼠模型和人类HSV-1感染病例中。HSV-1引起快速NETosis,由caspase-1激活和髓过氧化物酶分泌控制。中性粒细胞的明显积累,感染后在对侧眼保持未参与NET释放,暗示在未感染的眼睛中采取积极的防御姿势。此外,中性粒细胞耗竭加重眼部病理,病毒载量增加,和升级的疾病评分,证实NET在减少病毒复制中的保护作用。
■我们的报告揭示了NETosis以前未被探索的机制,通过对眼部HSV-1感染的促炎细胞死亡,和HPSE上调,为未来的研究寻找新的途径。
UNASSIGNED: Neutrophils are known mediators of innate immunity, yet their effector function in herpesvirus infections remains poorly understood. Here, we elucidate the mechanistic action and pivotal role of neutrophil extracellular traps (NETs) during herpes simplex virus type 1 (HSV-1) ocular infection.
UNASSIGNED: Neutrophils were collected from mice for HSV-1 infection, fluorescence imaging, and immunoblotting assay. Tear samples from healthy subjects and patients with HSV-1 and mice were collected at L. V. Prasad Eye Institute, India, and at the University of Illinois, USA, respectively. For the in vivo study, C57BL/6 mice as well as diversity outbred mice were infected with HSV-1 (McKrae strain) followed by tear fluid collection at various time points (0-10 days). Samples were used for Flow cytometry, ELISA, and immunofluorescence assay. Human transcriptomic profile of keratitis dataset was used evaluate NETosis signaling pathways. We also performed neutrophil depletion studies.
UNASSIGNED: Our data revealed a discernible temporal NET formation (NETosis) predominantly in the infected eye, across normal and diversity outbred murine models and human cases of HSV-1 infection. HSV-1 instigates swift NETosis governed by caspase-1 activation and myeloperoxidase secretion. Distinct accumulations of neutrophils, remaining unengaged in NET release in the contralateral eye post-infection, hinting at a proactive defensive posture in the uninfected eye. Moreover, neutrophil depletion accentuated ocular pathology, augmented viral load, and escalated disease scores, substantiating the protective effects of NETs in curtailing viral replication.
UNASSIGNED: Our report uncovers a previously unexplored mechanism of NETosis through pro-inflammatory cell death in response to ocular HSV-1 infection, and HPSE up-regulation, identifying new avenues for future studies.