越来越多的证据表明,细胞外囊泡(EV)是自身免疫性疾病中重要的细胞间信号转导,包括多发性硬化症(MS)。如果MS的病因仍然未知,它的分子生理学已经得到了很好的研究,表明外周血单核细胞(PBMC)是该疾病和神经炎症的主要病理相关贡献者。最近,多项研究表明,在中枢神经系统(CNS)自身免疫中,EV是神经免疫串扰的关键介质。为了评估电动汽车在MS中的作用,我们应用电子显微镜(EM)技术和Westernblot分析来研究血浆衍生的EV的形态和含量以及PBMC的超微结构,考虑四名MS患者和四名健康对照。通过其探索性,我们的研究能够检测到组间的显著差异.假足和大囊泡在病例的质膜界面较多,就像内质囊泡一样,导致PBMC的活化方面。此外,来自MS患者的PBMC还显示细胞质内的多囊泡体和囊泡周围的无定形物质的数量增加。此外,我们观察到大量的质膜覆盖的延伸,多个相关的大囊泡和许多含有未消化细胞质物质的自噬小体液泡。最后,对EV货物的研究证明了MS患者中许多失调的分子,包括GANAB,IFI35,Cortactin,Septin2,Cofilin1和ARHGDIA,在囊泡动力学改变的背景下作为炎症信号。我们得出的结论是,将EM与Western印迹分析应用于PBMC和囊泡化可以增强我们对MS病理生理学的了解。
Growing evidence identifies extracellular vesicles (EVs) as important cell-to-cell signal transducers in autoimmune disorders, including multiple sclerosis (MS). If the etiology of MS still remains unknown, its molecular physiology has been well studied, indicating peripheral blood mononuclear cells (
PBMCs) as the main pathologically relevant contributors to the disease and to neuroinflammation. Recently, several studies have suggested the involvement of EVs as key mediators of neuroimmune crosstalk in central nervous system (CNS) autoimmunity. To assess the role of EVs in MS, we applied electron microscopy (EM) techniques and Western blot analysis to study the morphology and content of plasma-derived EVs as well as the ultrastructure of
PBMCs, considering four MS patients and four healthy controls. Through its exploratory nature, our study was able to detect significant differences between groups. Pseudopods and large vesicles were more numerous at the plasmalemma interface of cases, as were endoplasmic vesicles, resulting in an activated aspect of the
PBMCs. Moreover,
PBMCs from MS patients also showed an increased number of multivesicular bodies within the cytoplasm and amorphous material around the vesicles. In addition, we observed a high number of plasma-membrane-covered extensions, with multiple associated large vesicles and numerous autophagosomal vacuoles containing undigested cytoplasmic material. Finally, the study of EV cargo evidenced a number of dysregulated molecules in MS patients, including GANAB, IFI35, Cortactin, Septin 2, Cofilin 1, and ARHGDIA, that serve as inflammatory signals in a context of altered vesicular dynamics. We concluded that EM coupled with Western blot analysis applied to
PBMCs and vesiculation can enhance our knowledge in the physiopathology of MS.