Viral Immunology Core

病毒免疫学核心

• 批准号: 10614636
• 负责人: OTTO O YANG
• 金额: $ 34.02万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-07 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614636
• 关键词: Address; Area; B-Lymphocytes; BLT mice; Biological Assay; Cell surface; Cells; Cellular Assay; Cellular Immunity; Clonality; Collaborations; Consult; Consultations; Cytomegalovirus; Data Analyses; Detection; Development; Engraftment; Enzyme-Linked Immunosorbent Assay; Evaluation; Experimental Designs; Flow Cytometry; Fostering; Funding; Genetic Engineering; Goals; HIV; HIV-1; Humoral Immunities; Immune; Immune response; Immunoglobulin Idiotypes; Immunologics; Immunology; Immunology procedure; Individual; Infection; Inflammatory; Intervention; Knowledge; Laboratories; Measurement; Methodology; Monitor; Mutation; Pathogenesis; Phenotype; Plasma; Research; Research Personnel; Running; Services; Standardization; Surface; T-Cell Proliferation; T-Lymphocyte; Testing; Transduction Gene; Transplantation; Viral; Viremia; antigen-specific T cells; antiviral immunity; cellular transduction; cytokine; deep sequencing; design; gene product; immune resistance; in vivo; integration site; latent HIV reservoir; mouse model; novel; operation; programs; reconstitution; success; tool; vector; virology

Core B: Project Summary/Abstract This core will provide support to the overall program by providing centralized viral and immunologic assays, as well as providing scientific guidance to each project. This will streamline, standardize, and integrate experimental methodology between Projects. There are two Aims: Aim 1. To perform immunologic and virologic assays in support of the interventions in the Projects, and develop or optimize novel assays as needed. • Flow cytometric analyses to quantify reconstitution in the huBLT mouse model • Flow cytometric identification of transduced cells expressing surface C46 • Phenotyping of transduced T and B cells • Evaluation of antigen-specific T cell proliferation by flow cytometry • Detection of bNAb expressing B cells by flow cytometry • Quantification of soluble bNAb (Design of quantitative ELISA) • Assessment of bNAb-directed anti-idiotype humoral immunity (Design of an anti-idiotype ELISA) • Monitoring for cellular immunity against transduced gene products • Monitoring for humoral immunity against cell surface transduced gene products C46 and/or huEGFRt (Design of a flow cytometric assay) • Measurement of plasma viremia in the huBLT mouse model • Antiviral assays for cellular immunity in the huBLT mouse model • Antiviral assays for bNAb produced in vivo • Sequencing of HIV-1 env for potential escape mutations • Quantitation of the latent reservoir in the huBLT mouse model • Testing functionality of CMV-specific CAR constructs • Measurement of pro-inflammatory cytokines • Clonal tracking of transduced cells by vector integration site deep sequencing Aim 2. To provide scientific and technical consultation to the Projects. The Core investigators will be available for scientific and technical consultation for each of the scientific projects.

核心B：项目总结/摘要 该核心将通过提供集中的病毒和免疫学检测，为整个项目提供支持， 并为每个项目提供科学指导。这将精简、标准化和整合 项目之间的实验方法。有两个目标： 目标1.进行免疫学和病毒学分析，以支持项目中的干预措施，以及 根据需要开发或优化新的检测方法。 ·流式细胞术分析以定量huBLT小鼠模型中的重建 ·表达表面C46的转导细胞的流式细胞术鉴定 ·转导的T和B细胞的表型分析 ·通过流式细胞术评估抗原特异性T细胞增殖 ·通过流式细胞术检测表达bNA B的B细胞 ·可溶性bNAb的定量（定量ELISA的设计） ·bNAb导向的抗独特型体液免疫的评估（抗独特型ELISA的设计） ·监测针对转导基因产物的细胞免疫 ·监测针对细胞表面转导的基因产物C46和/或huEGFR t的体液免疫 （流式细胞术测定的设计） ·测量huBLT小鼠模型中的血浆病毒血症 ·huBLT小鼠模型中细胞免疫的抗病毒测定 ·体内产生的bNAb的抗病毒测定 ·对HIV-1 env进行测序，以确定潜在的逃逸突变 ·huBLT小鼠模型中潜伏储库的定量 ·测试CMV特异性CAR构建体的功能 ·促炎细胞因子的测量 ·通过载体整合位点深度测序对转导细胞进行克隆追踪 目标2.为项目提供科学技术咨询。核心研究者将在 为每个科学项目提供科学和技术咨询。