Immune Monitoring Core

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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.