Allergy and Inmunology Department, Instituto de Investigaciones Sanitarias (ISS)-Fundación Jiménez Díaz, Universidad Autónoma de Madrid (UAM), Madrid, Spain.
Red de ASMA, REACCIONES ADVERSAS Y ALÉRGICAS (ARADyAL), Instituto de Salud Carlos III, Madrid, Spain.
Front Immunol. 2021 Jun 25;12:692569. doi: 10.3389/fimmu.2021.692569. eCollection 2021.
Anaphylaxis is a life-threatening systemic hypersensitivity reaction. During anaphylaxis, mediator release by effector cells causes endothelial barrier breakdown, increasing vascular permeability and leakage of fluids, which may lead to tissue edema. Although endothelial cells (ECs) are key players in this context, scant attention has been paid to the molecular analysis of the vascular system, and further analyses of this cell type are necessary, especially in humans. The protein expression pattern of human microvascular ECs was analyzed in response to sera from anaphylactic patients (EC-anaphylaxis) and sera from non-allergic subjects (EC-control) after 2 hours of contact. Firstly, a differential quantitative proteomic analysis of the protein extracts was performed by mass spectrometry using an isobaric labeling method. Second, the coordinated behavior of the identified proteins was analyzed using systems biology analysis (SBA). The proteome of the EC-anaphylaxis system showed 7,707 proteins, of which 1,069 were found to be significantly altered between the EC-control and EC-anaphylaxis groups (p-value < 0.05). Among them, a subproteome of 47 proteins presented a high rate of change (|ΔZq| ≥ 3). This panel offers an endothelial snapshot of the anaphylactic reaction. Those proteins with the highest individual changes in abundance were hemoglobin subunits and structural support proteins. The interacting network analysis of this altered subproteome revealed that the coagulation and complement systems are the main biological processes altered in the EC-anaphylactic system. The comprehensive SBA resulted in 5,512 functional subcategories (biological processes), 57 of which were significantly altered between EC-control and EC-anaphylaxis. The complement system, once again, was observed as the main process altered in the EC system created with serum from anaphylactic patients. Findings of the current study further our understanding of the underlying pathophysiological mechanisms operating in anaphylactic reactions. New target proteins and relevant signaling pathways operating in the endothelial-serum system have been identified. Interestingly, our results offer a protein overview of the micro-EC-anaphylaxis environment. The relevance of the coagulation, fibrinolytic, contact and complement systems in human anaphylaxis is described. Additionally, the untargeted high-throughput analysis used here is a novel approach that reveals new pathways in the study of the endothelial niche in anaphylaxis.
过敏反应是一种危及生命的全身性超敏反应。在过敏反应中，效应细胞释放介质会导致内皮屏障破裂，增加血管通透性和液体渗漏，从而导致组织水肿。尽管内皮细胞（ECs）是这方面的关键参与者，但对血管系统的分子分析关注甚少，需要进一步分析这种细胞类型，特别是在人类中。在与血清接触 2 小时后，分析了来自过敏患者（EC-过敏）和非过敏受试者（EC-对照）的血清对人微血管 ECs 的蛋白质表达模式。首先，使用等压标记法通过质谱对蛋白质提取物进行差异定量蛋白质组学分析。其次，使用系统生物学分析（SBA）分析鉴定蛋白的协调行为。EC-过敏系统的蛋白质组显示 7707 种蛋白质，其中 1069 种在 EC-对照和 EC-过敏组之间差异显著（p 值<0.05）。其中，47 种蛋白质的亚蛋白组表现出高变化率（|ΔZq|≥3）。该面板提供了过敏反应内皮快照。那些丰度变化最大的蛋白质是血红蛋白亚基和结构支持蛋白。对改变的亚蛋白组的相互作用网络分析表明，凝血和补体系统是改变的 EC-过敏系统中的主要生物学过程。全面的 SBA 产生了 5512 个功能亚类（生物学过程），其中 57 个在 EC-对照和 EC-过敏之间差异显著。补体系统再次被观察为来自过敏患者血清创建的 EC 系统中改变的主要过程。本研究的发现进一步加深了我们对过敏反应中潜在病理生理机制的理解。已确定新的靶蛋白和相关信号通路在内皮-血清系统中起作用。有趣的是，我们的结果提供了微 EC-过敏环境的蛋白质概述。描述了凝血、纤溶、接触和补体系统在人类过敏反应中的相关性。此外，这里使用的非靶向高通量分析是一种新方法，可在过敏反应内皮生态位的研究中揭示新途径。
