Immune Monitoring Core

摘要： 拟议的免疫监测核心(IMC；核心2)旨在提供统一、全面的 和集中服务，为U54调查人员提供高效和可重复的高通量分析 SARS-CoV-2抗体反应的研究，包括结合、功能特征和中和活性 (保留的临床样本和预期从核心1采集的血液样本)支持 项目1-3中所述的研究。IMC是CLIA认证的诊断分析平台的产物 由IMC调查人员开发，目前在埃默里医学实验室(EML)使用，以及 本系翻译研究实验室相关分析方法的发展。病理学和其他 埃默里遗址。IMC活动将由来自肿瘤学、病理学和 传染病、临床病理学家和训练有素的技术人员。该小组的努力将通过以下方式协调 三个项目、核心1和行政核心的领导层之间的定期会议。国际管理信息中心 活动将实现以下具体目标： 目的1.应用高通量自动RBD-EL ISA进行癌症和 风湿病患者在Emory Healthcare接受治疗。核心将利用抗RBD酶联免疫吸附试验 (由IMC研究人员开发)现已用于临床，以及该检测方法的变种，用于检测IgM和IgA 回应。这些化验方法将用于对目标保留的临床样本进行血清监测。 患者分组，以及预期收集的样本，如核心1中所述。 目的2.对新冠肺炎感染者进行详细的SARS-CoV-2抗体应答评估。 虽然自动化的酶联免疫吸附试验将被用来筛查所有预期收集的抗RBD抗体的样本 来自SARS-CoV-2感染患者的样本也将被检测抗SARS-CoV-2棘突抗体(S)， 核衣壳蛋白(N)、包膜蛋白(E)和膜蛋白(M)。此外，SARS的功能特征-- 冠状病毒-2抗体将包括抗体依赖的细胞介导的细胞毒性(ADCC)检测。 目的3.检测新冠肺炎感染者外周血中和能力。预期收集的样本 经EL ISA检测有SARS-CoV-2抗体的指定患者组，将在假病毒中进行检测 BSL2+条件下的中和试验。随后，那些阳性的样本将被确认 使用最近开发的焦点减少中和滴度(FRNT)-MNG分析。 总体而言，这一核心使U54研究人员能够利用最先进的诊断质量免疫 监控以实现项目目标。

Abstract: The proposed Immune Monitoring Core (IMC; Core 2) has been designed to provide a unified, comprehensive, and centralized service to provide the U54 investigators with efficient and reproducible high-throughput analysis of antibody responses to SARS-CoV-2, including binding, functional characterization, and neutralization activity (both retained clinical specimens and prospectively collected blood samples from Core 1) in support of the studies described in Projects 1-3. The IMC is an outgrowth of CLIA certified diagnostic assay platforms developed by IMC investigators and now in use in the Emory Medical Laboratories (EML), as well as a result of development of relevant assays in the Translational Research laboratory in the Dept. of Pathology and other Emory sites. IMC activities will be performed by experimental investigators from oncology, pathology, and infectious diseases, clinical pathologists, and trained technicians. This group’s efforts will be coordinated through regular meetings between leadership of the three Projects, Core 1, and the Administrative Core. The IMC activities will achieve the following Specific Aims: Aim 1. Employ the high-throughput automated RBD ELISA to perform serosurveillance of cancer and rheumatology patients treated within Emory Healthcare. The Core will utilize anti-RBD ELISA test (developed by IMC investigators) now in clinical use, as well variants of this test for detection of IgM and IgA responses. These assays will be used both for sero-surveillance on retained clinical samples from the target patient groups, and also on prospectively collected samples, as described in Core 1. Aim 2. Perform detailed evaluations of antibody response to SARS-CoV-2 in COVID-19 infected patients. While the automated ELISA will be used to screen all samples for anti-RBD antibodies, prospectively collected samples from SARS-CoV-2 infected patients will also be tested for antibodies against SARS-CoV-2 spike (S), nucleocapsid (N), envelope (E) and membrane (M) proteins. Additionally, functional characterization of SARS- CoV-2 antibodies will include assays of antibody dependent cell-mediated cytotoxicity (ADCC). Aim 3. Measuring the neutralization activity in COVID-19 infected patients. Prospectively collected samples from designated patient groups which have SARS-CoV-2 antibodies by ELISA will be tested in the pseudovirus neutralization assay under BSL2+ conditions. Subsequently, those samples that are positive will be confirmed using a recently developed focus reduction neutralization titer (FRNT)-mNG assay. Overall, this Core allows U54 investigators to take advantage of state-of-the-art and diagnostic quality immune monitoring to achieve Project goals.