Multi-omic immune profiling

摘要 我们提出了单细胞技术来量化多参数免疫标记物，这些标记物提供了- 用于分析我们研究队列中接种疫苗的免疫反应的深度单细胞数据。这个 此服务核心中的共享平台提供高效且可重现的配置文件，以支持我们的 具有极大的敏感性和细胞类型特异性的研究项目。定义单元的步骤 表型，我们将使用质量细胞术或CyTOF(飞行时间细胞测量法)来 多参数单细胞分析，它使用重金属离子作为抗体标记，并提供 为免疫亚群的细胞分析提供了极大的细节。为了进一步分析和关联 具有免疫表型的靶向单细胞的转录，我们将采用单细胞 循环血细胞转录组和表位测序索引(CITE-SEQ) 通过确定性条形码的空间分辨蛋白质转录组学(DBiT-seq) 组织(皮肤、淋巴结)。这些转录片段包括BCR和TCR的深度测序 研究组织中克隆性扩张与循环中的克隆性扩张。鉴定分子 在我们的队列中，我们将使用基于质谱仪(MS)的疫苗响应签名 基于MStern blotting的血清蛋白质组学平台无偏发现数百个 具有免疫功能的蛋白质很好地反映了免疫状态。 在化验和分析层面上的标准化将允许比较和整合 跨项目的数据。核心C的目标是提供独特且必要的资源 能够对血液和细胞的循环免疫细胞进行全面的表征 可从组织活检中获得，具有细胞飞行时间和RNA测序(RNA-SEQ)的比较数据， 和蛋白质组环境的匹配图谱。

Abstract We propose single-cell technologies to quantify multiparameter immune markers that provide in- depth single-cell data for analysis of immune responses to vaccination in our study cohorts. The shared platforms in this service core provide efficient and reproducible profiling in support of our research projects with greatly expanded sensitivity and cell-type specificity. To define cell phenotypes, we will employ mass cytometry or CyTOF (Cytometry by Time-Of-Flight) for multiparameter single-cell analysis, which uses heavy metal ions as antibody labels and provides tremendous detail for cellular analysis of immune subsets. To further profile and correlate the transcriptomes of targeted single cells with immunophenotype, we will employ single-cell cellular indexing of transcriptomes and epitopes by sequencing (CITE-Seq) for circulating blood cells and spatial-resolution proteo-transcriptomics via deterministic barcoding (DBiT-seq) in biopsies from tissues (skin, lymph node). These transcriptomics include in depth sequencing of BCR and TCR to investigate clonal expansion in the tissue compared to the circulation. To identify molecular signatures of vaccine responses across our cohorts, we will use a mass spectrometry (MS)-based MStern blotting-based serum proteomics platform for unbiased discovery of several hundred proteins with immunological function as an excellent reflection of the immunological state. Standardization, both at the assay and analytical levels, will allow comparisons and integration of data across projects. The goals of Core C are to provide a unique and essential resource to enable the comprehensive characterization of circulating immune cells from blood as well as cells available from tissue biopsies, with comparative data by CyTOF and RNA-sequencing (RNA-seq), and matched profiles of proteomic milieu.