Neutrophil extracellular traps induced by Haemonchus contortus excretory-secretory proteins varies among goats, gerbils, and mice.

IF 3.5 2区 医学 Q1 PARASITOLOGY
Yangchun Tan, Shuyi Cao, Muhammad Azhar Memon, Zhaohai Wen, Cheng Chen, Jiajun Feng, Xiaokai Song, Lixin Xu, Mingmin Lu, Ruofeng Yan
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引用次数: 0

Abstract

Background: Previous studies indicated that infection with Haemonchus contortus is host-specific (goat: susceptible host; gerbil: paratenic host; mouse: resistant host). Neutrophils play an essential role in host defense against parasitic infection through phagocytic engulfment, reactive oxygen species (ROS) generation, and neutrophil extracellular traps (NETs) formation. NETs are large web-like complexes consisting of a DNA scaffold decorated with various proteins components, including histones, myeloperoxidase, and elastase. They are released through both ROS-dependent and ROS-independent pathways. Previous studies have demonstrated both constraints and effectiveness of NETs in helminths. However, the roles of NETs in anti-infection of H. contortus in different hosts are still unclear.

Methods: To assess host-specific variations in NETs release, neutrophils isolated from goats, gerbils, and mice were co-cultured with Haemonchus contortus third-stage larvae (HcL3), followed by quantitative analysis of NETs formation using the PicoGreen® fluorescence assay. Subsequently, H. contortus excretory-secretory proteins (HcESPs) were co-cultured with neutrophils isolated from each host species. NETs release and ROS production were then quantitatively assessed using PicoGreen® fluorescence intensity and oxidation-sensitive dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescence. In addition, the neutrophil's phagocytic ability for FITC-dextran was evaluated by flow cytometric analysis. Finally, to elucidate the signaling pathways involved in HcESP-induced NETs release in goat neutrophils, four specific inhibitors were employed for pretreatment prior to stimulation.

Results: Our results demonstrate that in vitro stimulation with HcL3 triggers NETs formation. The release of NETs exhibits significant host-specific variation, specifically, neutrophils from mice showed the highest NETs release, followed by gerbils, and a minimal response in goats. Moreover, HcESP treatment markedly inhibited ROS generation and phagocytic capacity in neutrophils from all three host species. Intriguingly, HcESPs exerted host-specific modulation of NETs release, with inhibition observed in goats, enhancement in mice, and context-dependent modulation in gerbils. Mechanistic investigations revealed that the NETs suppression in goats neutrophils involved both nicotinamide adenine dinucleotide phosphate (NADPH) oxidase- and neutrophil elastase-dependent pathways.

Conclusions: Our results demonstrate that HcESPs significantly inhibit NETs formation in goat neutrophils through dual modulation of NADPH oxidase and neutrophil elastase activity. This finding highlights these two enzymes as promising molecular targets for anti-helminthic vaccine development.

山羊、沙鼠和小鼠的中性粒细胞胞外陷阱是由弯曲血蜱的排泄-分泌蛋白引起的。
背景:以往的研究表明,弯曲血蜱的感染是宿主特异性的(山羊:易感宿主;沙鼠:副毒宿主;老鼠:抗宿主)。中性粒细胞通过吞噬吞噬、活性氧(ROS)的产生和中性粒细胞胞外陷阱(NETs)的形成,在宿主防御寄生虫感染中发挥重要作用。net是由DNA支架组成的大型网状复合体,支架上装饰着各种蛋白质成分,包括组蛋白、髓过氧化物酶和弹性酶。它们通过ros依赖性和ros非依赖性途径释放。以前的研究已经证明了蚊帐对蠕虫的限制和有效性。然而,NETs在不同宿主中抗弯纹弓形虫感染的作用尚不清楚。方法:为了评估宿主释放NETs的特异性变化,将山羊、沙鼠和小鼠中分离的中性粒细胞与弯血蜱第三期幼虫(HcL3)共培养,然后使用PicoGreen®荧光法定量分析NETs的形成。随后,将H. contortus的排泄分泌蛋白(HcESPs)与从每种宿主中分离的中性粒细胞共培养。然后使用PicoGreen®荧光强度和氧化敏感的二氯二氢荧光素(DCFH-DA)荧光定量评估NETs释放和ROS生成。此外,用流式细胞术分析中性粒细胞对fitc -葡聚糖的吞噬能力。最后,为了阐明hcsp诱导的山羊中性粒细胞释放NETs的信号通路,在刺激前使用了四种特异性抑制剂进行预处理。结果:我们的研究结果表明,HcL3体外刺激可触发NETs的形成。NETs的释放表现出显著的宿主特异性差异,具体来说,小鼠的中性粒细胞释放的NETs最高,其次是沙鼠,山羊的反应最低。此外,HcESP处理显著抑制了所有三种宿主的中性粒细胞的ROS生成和吞噬能力。有趣的是,HcESPs发挥宿主特异性调节NETs释放的作用,在山羊中观察到抑制作用,在小鼠中观察到增强作用,在沙鼠中观察到环境依赖性调节作用。机制研究表明,山羊中性粒细胞的NETs抑制涉及烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶和中性粒细胞弹性酶依赖途径。结论:我们的研究结果表明,HcESPs通过双重调节NADPH氧化酶和中性粒细胞弹性酶活性,显著抑制山羊中性粒细胞NETs的形成。这一发现强调了这两种酶是抗蠕虫疫苗开发的有希望的分子靶点。
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来源期刊
Parasites & Vectors
Parasites & Vectors 医学-寄生虫学
CiteScore
6.30
自引率
9.40%
发文量
433
审稿时长
1.4 months
期刊介绍: Parasites & Vectors is an open access, peer-reviewed online journal dealing with the biology of parasites, parasitic diseases, intermediate hosts, vectors and vector-borne pathogens. Manuscripts published in this journal will be available to all worldwide, with no barriers to access, immediately following acceptance. However, authors retain the copyright of their material and may use it, or distribute it, as they wish. Manuscripts on all aspects of the basic and applied biology of parasites, intermediate hosts, vectors and vector-borne pathogens will be considered. In addition to the traditional and well-established areas of science in these fields, we also aim to provide a vehicle for publication of the rapidly developing resources and technology in parasite, intermediate host and vector genomics and their impacts on biological research. We are able to publish large datasets and extensive results, frequently associated with genomic and post-genomic technologies, which are not readily accommodated in traditional journals. Manuscripts addressing broader issues, for example economics, social sciences and global climate change in relation to parasites, vectors and disease control, are also welcomed.
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