Viral Immunology Core

核心B：项目摘要/摘要 该核心将通过提供集中化的病毒和免疫学检测为整个计划提供支持，如 并为每个项目提供科学指导。这将简化、标准化和集成 项目间的实验方法。有两个目标： 目的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分析方法。 ·流式细胞仪分析以量化huBLT小鼠模型中的重组 ·流式细胞仪鉴定表达表面C46的转导细胞 ·转导的T和B细胞的表型 ·用流式细胞术评价抗原特异性T细胞的增殖 ·流式细胞术检测表达bNAb的B细胞 ·可溶性bNAb的定量(定量酶联免疫吸附试验设计) ·评估bNAb导向的抗独特型体液免疫(设计抗独特型酶联免疫吸附试验) ·监测针对转导基因产品的细胞免疫 ·监测针对细胞表面转导基因产物C46和/或huEGFRt的体液免疫 (一种流式细胞仪的设计) ·在huBLT小鼠模型中测量血浆病毒血症 ·huBLT小鼠模型细胞免疫的抗病毒检测 ·体内生产的bNAb的抗病毒检测 ·对HIV-1包膜病毒进行测序，以寻找潜在的逃逸突变 ·在huBLT小鼠模型中对潜伏水库进行量化 ·测试CMV专用汽车结构的功能 ·促炎细胞因子的测定 ·利用载体整合位点深度测序技术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。核心调查员将有空 为每个科学项目提供科学技术咨询。

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.