Proteomics Core Facility, Istituto Superiore di Sanità(ISS), Rome, Italy.
Who Collaborating Centre for Epidemiology, Detection and Control of Cystic and Alveolar Echinococcosis, Department of Infectious, Diseases (DMI), ISS, Rome, Italy.
PLoS Negl Trop Dis. 2020 Oct 5;14(10):e0008586. doi: 10.1371/journal.pntd.0008586. eCollection 2020 Oct.
The reference diagnostic method of human abdominal Cystic Echinococcosis (CE) is imaging, particularly ultrasound, supported by serology when imaging is inconclusive. However, current diagnostic tools are neither optimal nor widely available. The availability of a test detecting circulating biomarkers would considerably improve CE diagnosis and cyst staging (active vs inactive), as well as treatments and follow-up of patients. Exosomes are extracellular vesicles involved in intercellular communication, including immune system responses, and are a recognized source of biomarkers. With the aim of identifying potential biomarkers, plasma pools from patients infected by active or inactive CE, as well as from control subjects, were processed to isolate exosomes for proteomic label-free quantitative analysis. Results were statistically processed and subjected to bioinformatics analysis to define distinct features associated with parasite viability. First, a few parasite proteins were identified that were specifically associated with either active or inactive CE, which represent potential biomarkers to be validated in further studies. Second, numerous identified proteins of human origin were common to active and inactive CE, confirming an overlap of several immune response pathways. However, a subset of human proteins specific to either active or inactive CE, and central in the respective protein-protein interaction networks, were identified. These include the Src family kinases Src and Lyn, and the immune-suppressive cytokine TGF-β in active CE, and Cdc42 in inactive CE. The Src and Lyn Kinases were confirmed as potential markers of active CE in totally independent plasma pools. In addition, insights were obtained on immune response profiles: largely consistent with previous evidence, our observations hint to a Th1/Th2/regulatory immune environment in patients with active CE and a Th1/inflammatory environment with a component of the wound healing response in the presence of inactive CE. Of note, our results were obtained for the first time from the analysis of samples obtained in vivo from a well-characterized, large cohort of human subjects.
人类腹部包虫病(CE)的参考诊断方法是影像学,尤其是超声检查,当影像学检查结果不确定时,辅以血清学检查。然而,目前的诊断工具既不理想也不广泛可用。检测循环生物标志物的检测方法的出现将极大地改善 CE 的诊断和囊肿分期(活动与非活动),以及患者的治疗和随访。外泌体是参与细胞间通讯的细胞外囊泡,包括免疫系统反应,是生物标志物的公认来源。为了确定潜在的生物标志物,对来自活动性或非活动性 CE 感染患者以及对照者的血浆池进行处理,以分离外泌体进行无标记蛋白质组定量分析。结果进行了统计学处理,并进行了生物信息学分析,以确定与寄生虫活力相关的独特特征。首先,鉴定出一些与活动性或非活动性 CE 特异性相关的寄生虫蛋白,这些蛋白代表了有待进一步研究验证的潜在生物标志物。其次,鉴定出的许多人类来源的蛋白在活动性和非活动性 CE 中是共同的,证实了几种免疫反应途径的重叠。然而,还鉴定出了一组与活动性或非活动性 CE 特异性相关的人类蛋白,并且在各自的蛋白质-蛋白质相互作用网络中处于核心地位。这些蛋白包括Src 家族激酶Src 和 Lyn,以及在活动性 CE 中具有免疫抑制作用的细胞因子 TGF-β,以及在非活动性 CE 中具有 Cdc42。Src 和 Lyn 激酶在完全独立的血浆池中被证实为活动性 CE 的潜在标志物。此外,还获得了关于免疫反应谱的见解:与之前的证据基本一致,我们的观察结果暗示在活动性 CE 患者中存在 Th1/Th2/调节性免疫环境,而在非活动性 CE 中存在 Th1/炎症环境,其中包含伤口愈合反应的成分。值得注意的是,我们的结果是首次从对来自特征明确的大型人类受试者队列的体内样本进行的分析中获得的。