Capriotti Lorena, Iuliano Marco, Lande Roberto, Frasca Loredana, Falchi Mario, Rosa Paolo, Mangino Giorgio, Romeo Giovanna
Department of Medico-Surgical Sciences and Biotechnologies, Sapienza University of Rome - Polo Pontino, Latina, Italy.
Pharmacological Research and Experimental Therapy Section, National Center for Drug Research and Evaluation, Istituto Superiore di Sanità, Rome, Italy.
J Inflamm Res. 2022 Sep 16;15:5387-5399. doi: 10.2147/JIR.S373150. eCollection 2022.
Extracellular Vesicles (EVs) are a heterogeneous group of cell-derived membranous nanoparticles involved in several physiopathological processes. EVs play a crucial role in the definition of the extracellular microenvironment through the transfer of their cargo. Psoriasis is a prototypical chronic inflammatory disease characterized by several secreted mediators, among which antimicrobial peptides (AMPs) are considered pivotal in the development of the psoriatic inflammatory microenvironment. The role of EVs in the pathogenesis of psoriasis has not been elucidated yet, even if emerging evidence demonstrated that interleukin-17A (IL-17A), the psoriasis-related principal cytokine, modifies EVs release and cargo content. The aim of this work was to analyze whether, besides IL-17A, other psoriasis-related cytokines (ie, IFN-γ, TNF-α, IL-22 and IL-23) could affect EVs release and their AMPs mRNAs cargo as well as to analyze the potential biological effect due to EVs internalization by different acceptor cells.
Nanoparticle tracking analysis (NTA) was performed on supernatants of HaCaT cells stimulated with IL-17A, IFN-γ, TNF-α, IL-22 or IL-23 to enumerate EVs. Real-Time RT-PCR was used for gene expression analysis in cells and EVs. Confocal microscopy and Flow cytometry were used to, respectively, study Netosis and EVs internalization.
IL-17A and IFN-γ increased EVs release by HaCaT cells. All the tested cytokines modulated AMPs mRNA expression in parental cells and in their respective EVs. and mRNAs were upregulated following IL-17A and IL-22 treatments. Interestingly, EVs derived from cytokine treated HaCaT cells induced Netosis in freshly isolated neutrophils. Upregulation of and mRNA was also detectable in acceptor cells incubated with EVs derived from cells treated with psoriasis-related cytokines.
The obtained results highlighted the role of EVs in the composition of psoriasis-associated secretome and microenvironment also suggesting the EV involvement in the spreading of the disease mediators and in the possible associated comorbidities.
细胞外囊泡（EVs）是一组异质性的细胞来源的膜性纳米颗粒，参与多种生理病理过程。EVs通过其内容物的传递在细胞外微环境的形成中发挥关键作用。银屑病是一种典型的慢性炎症性疾病，其特征是多种分泌介质，其中抗菌肽（AMPs）被认为在银屑病炎症微环境的发展中起关键作用。尽管有新证据表明银屑病相关的主要细胞因子白细胞介素-17A（IL-17A）可改变EVs的释放和内容物，但EVs在银屑病发病机制中的作用尚未阐明。这项工作的目的是分析除IL-17A外，其他银屑病相关细胞因子（即IFN-γ、TNF-α、IL-22和IL-23）是否会影响EVs的释放及其AMPs mRNA内容物，并分析不同受体细胞内化EVs所产生的潜在生物学效应。
对用IL-17A、IFN-γ、TNF-α、IL-22或IL-23刺激的HaCaT细胞上清液进行纳米颗粒跟踪分析（NTA）以计数EVs。实时逆转录聚合酶链反应（Real-Time RT-PCR）用于细胞和EVs中的基因表达分析。共聚焦显微镜和流式细胞术分别用于研究细胞焦亡和EVs内化。
IL-17A和IFN-γ增加了HaCaT细胞的EVs释放。所有测试的细胞因子均调节亲代细胞及其各自EVs中AMPs mRNA的表达。IL-17A和IL-22处理后，和mRNA上调。有趣的是，细胞因子处理的HaCaT细胞衍生的EVs在新鲜分离的中性粒细胞中诱导细胞焦亡。在用银屑病相关细胞因子处理的细胞衍生的EVs孵育的受体细胞中，也可检测到和mRNA的上调。
所得结果突出了EVs在银屑病相关分泌组和微环境组成中的作用，也表明EVs参与疾病介质的传播以及可能的相关合并症。
