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.

利用高色流式细胞术检测医疗器械开发和传染病中的免疫激活和极化 我们已经在人类样本上使用了高色流式细胞术，目前正在为小鼠样本设计抗体组。 FACS 中的大多数抗体面板在标准仪器上仅限于 15 种颜色。通过利用光谱细胞术，我们目前正在开发 37 色人类流式细胞术面板，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）和儿科扁桃体样本（合作：Schwartzberg）中的 COVID-19 研究中实施。该面板还将用于 NHTSA 全年收集的 700 多个创伤样本，以表征创伤受害者的 PBMC 反应，并与 COVID 抗体血清阳性状态相关联。我们正在优化两个超过 30 种颜色的抗体组的设计，用于对小鼠模型中的骨髓细胞和淋巴细胞进行表型分析。这些将在 2020 财年末至 2021 财年年初在实验室实施和测试。 开发用于检测 SARS-CoV-2 抗体的高通量血清学检测方法 为了帮助了解 SARS-CoV-2 大流行，我们的实验室开发了一种特异性且灵敏的检测方法来评估健康成人的血清阳性率。我们与弗雷德里克国家癌症研究实验室、国家转化科学促进中心和国家过敏和传染病研究所合作进行了下述血清学调查。我们的检测开发目标是创建一种简单、经济实惠、易于使用和实施的检测方法，不需要特殊设备或试剂。因此，我们开发了具有 100% 敏感性（95% CI：94.48% 至 100%）和 100% 特异性（95% CI：98.78% - 100%）的酶联免疫吸附测定 (ELISA)，并作为向 FDA 申请紧急使用授权。 设计并启动全国血清调查，以评估美国未确诊成年人中 COVID-19 的传播情况 利用我们新设计的 ELISA，我们试图通过血清学调查来评估未诊断出 SARS-CoV-2 感染但之前接触过该病毒的健康成年人的比例。我们与 NCATS、NIAID 和 FNLCR 合作，通过邮寄采血设计了一项全国血清调查。经过广泛招募志愿者后，我们收到了 40 万名潜在捐赠者。然后，这些志愿者提供了他们的基本人口统计信息，这些信息被输入到算法中，该算法根据最近的人口普查数据选择了代表美国人口的 10,000 名捐赠者。这 10,000 名捐献者通过电话获得同意，然后运送他们在家中完成的血液采样套件，随后运送到马里兰州贝塞斯达的实验室。使用上述测定法处理样品并测试抗 SARS-CoV-2 IgG、IgM 和 IgA 抗体。然后将这些实验室数据与临床问卷结合起来，分析与 SARS-CoV-2 血清阳性相关的因素。我们现在正在收集最后的样本，希望在 2020 年 9 月底之前完成研究。