Immunoengineering - Supplemental Funding

免疫工程 - 补充资金

• 批准号: 10291096
• 负责人: Kaitlyn Sadtler
• 金额: $ 38万
• 依托单位: NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10291096
• 关键词: 2019-nCoV; Adult; Algorithms; Animals; Antibodies; Authorization documentation; Biocompatible Materials; Biological; Biological Assay; Blood specimen; COVID-19; Censuses; Childhood; Clinical; Collaborations; Color; Communicable Diseases; Communities; Consent; Cytometry; Data; Detection; Development; Device or Instrument Development; Diagnosis; Emergency Situation; Enzyme-Linked Immunosorbent Assay; Equipment; Exposure to; Flow Cytometry; Funding; Goals; Home environment; Human; Immune response; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Infection; Laboratories; Lymphoid Cell; Lymphoid Tissue; Medical Device; Mus; Myeloid Cells; National Institute of Allergy and Infectious Disease; National Institute of Biomedical Imaging and Bioengineering; Peripheral Blood Mononuclear Cell; Phenotype; Process; Protocols documentation; Questionnaires; Reagent; Regenerative Medicine; Sampling; Serological; Ships; Specificity; Surveys; Telephone; Testing; Tonsil; Translational Research; Trauma; United States; United States National Institutes of Health; Virus; anticancer research; assay development; coronavirus disease; design; immune activation; immune function; immunoengineering; implantable device; implantation; instrument; mouse model; pandemic disease; peripheral blood; population based; process optimization; programs; response; seropositive; tool; volunteer

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.

利用高色流式细胞仪在医疗器械发展和传染病中审问免疫激活和极化 我们已经在人类样品上实施了高色流式细胞仪的使用，目前正在设计用于小鼠样品的抗体面板。 FACS中的大多数抗体面板在标准仪器上仅限于15种颜色。通过利用光谱细胞仪，我们目前正在使用一个37色的人流式细胞仪面板前进，用于外周血单核细胞和中央淋巴组织。该小组正在成人血液样本（合作：Memoli，Hall，Esposito）和小儿扁桃体样品（合作：Schwartzberg）中实施。该面板还将用于NHTSA全年收集的一组700多个创伤样本，以表征创伤受害者中的PBMC反应，并与共同的抗体血清阳性状态相关。我们正在优化两种> 30个颜色抗体面板的设计，用于在小鼠模型中表型和淋巴样细胞。这些将在2020财年后期之前在实验室中实施和测试，直到2021财年开始。 开发用于检测SARS-COV-2抗体的高通量血清学测定 为了帮助理解SARS-COV-2大流行，我们的实验室开发了一种特定敏感的测定法，以评估健康成年人的血清阳性。我们与弗雷德里克国家癌症研究实验室，国家前进的转化科学中心和国家过敏和传染病研究所合作，以下面介绍的血清学调查。我们的测定开发目标是创建一个简单，负担得起的，易于使用和实施的测定，不需要利基设备或试剂。因此，我们具有100％敏感性（95％CI：94.48％至100％）和100％的特异性（95％CI：98.78％-100％），以100％的敏感性（95％CI：94.48％至100％）开发和酶联免疫吸附测定法（ELISA）。 全国血清监管的设计和开始，以评估美国在美国未诊断的成年人中的covid-19 利用我们新设计的ELISA，我们试图评估尚未诊断出患有SARS-COV-2感染的健康成年人的比例，而SARS-COV-2感染以前通过血清学调查暴露于病毒。与NCAT，NIAID和FNLCR合作，我们通过邮寄血液采样设计了全国血清调查。在召集志愿者的一般呼吁之后，我们收到了40万个潜在的捐助者。然后，这些志愿者提供了他们的基本人口统计信息，该信息被送入了一种算法，该算法选择了10,000名捐助者，该捐助者根据最近的人口普查数据代表了美国人口。这10,000名捐赠者通过电话同意，然后运送了他们在家完成的血液采样套件，然后运送到马里兰州贝塞斯达的实验室。对样品进行处理并测试，并使用上述测定法对抗SARS-COV-2 IgG，IgM和IgA抗体进行了测试。然后将该实验室数据与临床问卷结合使用，以分析与SARS-COV-2血清阳性相关的因素。我们正在收集最后的样本，并希望在2020年9月底之前完成研究。