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Principal Investigator: Kaitlyn Sadtler
Organization: NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING
Fiscal Year: 2020
Award: $379,950
Funding agency: National Institute of Biomedical Imaging and Bioengineering
Utilization of High-Color Flow Cytometry for Interrogation of Immune Activation and Polarization in Medical Device Development and Infectious Disease
We have implemented the use of high-color flow cytometry on human samples and are currently designing antibody panels for mouse samples. Most antibody panels in FACS are limited to 15 colors on standard instruments. By utilizing spectral cytometry, we are currently moving forward with a 37-color human flow cytometry panel for use on peripheral blood mononuclear cells and central lymphoid tissues. This panel is being implemented in the study of COVID-19 in adult blood samples (collab: Memoli, Hall, Esposito) and pediatric tonsil samples (collab: Schwartzberg). This panel will also be utilized against a set of 700+ trauma samples that have been collected throughout the year by the NHTSA to characterize PBMC responses in trauma victims and correlate with COVID antibody seropositivity status. We are in the process of optimizing the design of two >30 color antibody panels for phenotyping myeloid and lymphoid cells in mouse models. These will be implemented and tested in the lab in late FY2020 into the beginning of FY2021.
Development of High-Throughput Serologic Assays for Detection of SARS-CoV-2 Antibodies
To assist in the understanding of the SARS-CoV-2 pandemic, our laboratory developed a specific and sensitive assay to evaluate seropositivity in healthy adults. We partnered with Frederick National Laboratory for Cancer Research, the National Center for Advancing Translational Sciences and the National Institute for Allergy and Infectious Diseases for the serologic survey described below. Our goals for assay development were to create a simple, affordable, easily used and implemented assay that didnt require niche equipment or reagents. As such, we developed and enzyme-linked immunosorbent assay (ELISA) with 100% sensitivity (95% CI: 94.48% to 100%) and 100% specificity (95% CI: 98.78% - 100%) that is being submitted as an application for Emergency Use Authorization from the FDA.
Design and commencement of a national serosurvey for evaluating the spread of COVID-19 in undiagnosed adults in the United States
Utilizing our newly designed ELISAs, we sought to evaluate the proportion of healthy adults that had not been diagnosed with SARS-CoV-2 infection that had previously been exposed to the virus through a serologic survey. In collaboration with NCATS, NIAID, and FNLCR, we designed a national serosurvey through mail-in blood sampling. After a general call for volunteers, we received 400,000 potential donors. These volunteers then provided their basic demographic information that was fed into an algorithm that selected 10,000 donors that represented the US Population based on recent census data. These 10,000 donors were consented over the phone, then shipped a blood sampling kit that they completed at home, and subsequently shipped to the laboratory in Bethesda, MD. Samples were processed and tested for anti-SARS-CoV-2 IgG, IgM, and IgA antibodies with the assays described above. This laboratory data was then combined with a clinical questionnaire to analyze factors that are correlated with SARS-CoV-2 seropositivity. We are in the process of collecting the last samples now and hope to have the study completed by the end of September 2020.
Terms: <19S Gamma Globulin><2019 novel coronavirus><2019-nCoV><21+ years old><7S Gamma Globulin><Adult><Adult Human><Algorithms><Animals><Antibodies><Assay><Authorization><Authorization documentation><Bioassay><Biocompatible Materials><Biologic Assays><Biological><Biological Assay><Biomaterials><Blood Sample><Blood specimen><COVID><COVID-19><COVID19><Cellular Immune Function><Censuses><Childhood><Clinical><CoV disease><Collaborations><Color><Communicable Diseases><Communities><Consent><Cytometry><Data><Detection><Development><Device or Instrument Development><Diagnosis><ELISA><Emergencies><Emergency Situation><Enzyme-Linked Immunosorbent Assay><Equipment><Exposure to><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Funding><Goals><Home><Home environment><Human><IgA><IgG><IgM><Immune Cell Activation><Immune response><Immunoglobulin A><Immunoglobulin G><Immunoglobulin M><Immunological response><Infection><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Laboratories><Lymphatic Tissue><Lymphoid Cell><Lymphoid Tissue><Medical Device><Mice><Mice Mammals><Modern Man><Murine><Mus><Myeloid Cells><NIAID><NIBIB><NIH><National Institute of Allergy and Infectious Disease><National Institute of Biomedical Imaging and Bioengineering><National Institutes of Health><PBMC><Peripheral Blood Mononuclear Cell><Permission><Phenotype><Phone><Process><Protocol><Protocols documentation><Questionnaires><Reagent><Regenerative Medicine><SARS-CoV-2><SARS-CoV2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related coronavirus 2><Sampling><Serologic><Serological><Severe acute respiratory syndrome coronavirus 2><Ships><Specificity><Survey Instrument><Surveys><Telephone><Testing><Tonsil><Translational Research><Translational Science><Trauma><United States><United States National Institutes of Health><Virus><Wuhan coronavirus><adulthood><anti-cancer research><anticancer research><assay development><biological material><biomedical implant><cancer research><corona virus disease><corona virus disease 2019><coronavirus disease><coronavirus disease 2019><design><designing><developmental><device development><engineered immune system><flow cytophotometry><host response><immune activation><immune engineering><immune function><immunoengineering><immunoresponse><implant device><implantable device><implantation><indwelling device><instrument><instrument development><mouse model><murine model><pandemic><pandemic disease><pediatric><peripheral blood><population based><process optimization><programs><response><seropositive><tonsillar><tool><translation research><volunteer>