Resolution of inflammation after cardiac arrest

摘要 院外心脏骤停会引起早期全身炎症反应，与 严重的神经损伤和死亡率。与脓毒症不同的是，uchA提供了一种干预的机会 免疫反应的早期阶段。然而，心脏骤停的免疫学研究还不够深入。至 为了解决这一问题，我们启动了多中心心脏骤停免疫学网络(I-CAN)，这是一项合作 应用单细胞方法建立一种独特的超低温保存的外周血单核细胞生物库 外周血细胞(PBMC)。我们的前提是心脏骤停触发内源性、代偿性 促进炎症消退、限制损伤和提高存活率的机制。我们的预赛 研究证实，患者中表达免疫检查点的单核细胞和NK细胞状态扩大 神经学结果很差。交互作用组分析确定了介导串音的细胞因子(干扰素γ、白介素10) Nectin-2+单核细胞与Tim-3+TIGIT+NK细胞之间的关系。细胞色素A体外培养PBMC的研究进展 研究表明，Nectin-2对NK细胞产生干扰素γ有抑制作用。干扰素γ基因缺陷小鼠的实验研究 实验性心脏骤停和复苏减少了神经损伤和死亡率。这些发现 提示我们的假设是Nextin-2单核细胞是一种保护性反应，以减轻炎症和 神经性损伤。在这里，我们建议定义消退炎症的机制 心脏骤停。在目标1中，我们在单细胞分辨率下对uchA患者进行了深度免疫表型分析。在……里面 目的2，我们定义了uchA的免疫检查点特征及其与神经预后的关系。我们 然后定义免疫检查点的功能，重点是Nectin-2+单核细胞。在目标3中，我们 在心脏骤停和心脏骤停小鼠模型中测试免疫检查点受体作为治疗策略的靶向性 复苏。这些目标共同定义了一种新的治疗方法：增强内源性、 减少心脏骤停后炎症的保护性机制。此外，我们的多中心和多中心 心脏骤停免疫学网络(I-CAN)的学科小组建立了两个站点的资源 临床心脏骤停的生物库和受试者的深层免疫表型。

Abstract Out of hospital cardiac arrest (OHCA) produces an early systemic inflammatory response associated with significant neurological injury and mortality. Unlike sepsis, OHCA offers an opportunity to intervene at the earliest stages of the immune response. However, the immunology of cardiac arrest is understudied. To address this, we started the multi-center Immunology of Cardiac Arrest Network (I-CAN), a collaboration applying single-cell approaches to a unique biobank of cryopreserved, viable peripheral blood mononuclear cells (PBMC) from OHCA patients. Our premise is that cardiac arrest triggers endogenous, compensatory mechanisms that promote the resolution of inflammation, limit injury and improve survival. Our preliminary studies identified that monocyte and NK cell states expressing immune checkpoints were expanded in patients with poor neurological outcomes. Interactome analysis identified cytokines (IFNγ, IL-10) that mediate cross-talk between Nectin-2+ monocytes and Tim-3+ TIGIT+ NK cells. Subsequent ex vivo studies on PBMC from OHCA patients demonstrated that Nectin-2 is a brake on production of IFNγ by NK cells. IFNγ-deficient mice in experimental cardiac arrest and resuscitation had reduced neurological injury and mortality. These findings suggest our hypothesis that Nectin-2 monocytes are a protective response to ameliorate inflammation and neurological injury after OHCA. Here, we propose to define the mechanisms that resolve inflammation after cardiac arrest. In Aim 1, we perform deep immunophenotyping of OHCA patients at single-cell resolution. In Aim 2, we define the immune checkpoint profile of OHCA and its association with neurological outcomes. We then define the function of immune checkpoints in OHCA, with a focus on Nectin-2+ monocytes. In Aim 3, we test targeting of immune checkpoint receptors as a therapeutic strategy in a mouse model of cardiac arrest and resuscitation. Together, these aims define a new therapeutic approach: augmentation of endogenous, protective mechanisms to reduce inflammation after cardiac arrest. Further, our multi-center and multi- disciplinary team in the Immunology of Cardiac Arrest Network (I-CAN) establishes the resource of a two-site biorepository of clinical cardiac arrest with deep immunophenotyping of the subjects.