PDF(Pubmed)
Abstract：
Extracellular vesicles (EVs) are membrane-enclosed nanoparticles that transport several biomolecules and are involved in important mechanisms and functions related to the pathophysiology of fungal diseases. EVs from Paracoccidioides brasiliensis, the main causative agent of Paracoccidioidomycosis (PCM), modulate the immune response of macrophages. In this study, we assessed the EVs proteome from a virulent P. brasiliensis isolated from granulomatous lesions and compared their immunomodulatory ability with EVs isolated from the fungus before the animal passage (control EVs) when challenging macrophages and dendritic cells (DCs). Proteome showed that virulent EVs have a higher abundance of virulence factors such as GP43, protein 14-3-3, GAPDH, as well as virulence factors never described in PCM, such as aspartyl aminopeptidase and a SidJ analogue compared with control EVs. Virulent extracellular vesicles induced higher expression of TLR4 and Dectin-1 than control EVs in macrophages and dendritic cells (DCs). In opposition, a lower TLR2 expression was induced by virulent EVs. Additionally, virulent EVs induced lower expression of CD80, CD86 and TNF-α, but promoted a higher expression of IL-6 and IL-10, suggesting that EVs isolated from virulent P. brasiliensis-yeast promote a milder DCs and macrophage maturation. Herein, we showed that EVs from virulent fungi stimulated a higher frequency of Th1/Tc1, Th17, and Treg cells, which gives new insights into fungal extracellular vesicles. Taken together, our results suggest that P. brasiliensis utilizes its EVs as virulence bags that manipulate the immune system in its favour, creating a milder immune response and helping with fungal evasion from the immune system.
摘要：
细胞外囊泡（EV）是膜封闭的纳米颗粒，可运输几种生物分子，并参与与真菌疾病的病理生理学相关的重要机制和功能。来自巴西副球菌的电动汽车，副孢子菌病（PCM）的主要病原体，调节巨噬细胞的免疫反应。在这项研究中,我们评估了从肉芽肿性病变中分离出的毒力巴西化验菌的EV蛋白质组,并比较了它们与动物传代前从真菌中分离出的EV(对照EV)在攻击巨噬细胞和树突状细胞(DC)时的免疫调节能力.蛋白质组显示,强毒力EV具有较高丰度的GP43、蛋白14-3-3、GAPDH、以及PCM中从未描述过的毒力因子，例如天冬氨酰氨基肽酶和SidJ类似物与对照电动汽车相比。在巨噬细胞和树突状细胞(DC)中,与对照EV相比,强的细胞外囊泡诱导更高的TLR4和Dectin-1表达。在反对中，毒性EV诱导较低的TLR2表达。此外,强毒电动汽车诱导CD80、CD86和TNF-α表达降低，但促进了IL-6和IL-10的较高表达，表明从强毒巴西酵母中分离出的EV促进了温和的DC和巨噬细胞成熟。在这里，我们发现，来自毒性真菌的EV刺激更高频率的Th1/Tc1，Th17和Treg细胞，这为真菌细胞外囊泡提供了新的见解。一起来看,我们的结果表明，巴西假单胞菌利用其电动汽车作为毒力袋，操纵免疫系统对其有利，创造更温和的免疫反应，帮助真菌逃避免疫系统。
