High-resolution single cell profiling of vaccine responsiveness in the elderly

项目概要 该项目的目标是了解免疫系统中与衰老相关的基因组和功能变化 影响流感疫苗接种反应的细胞。衰老的免疫系统的能力下降 对疫苗的保护性反应是对老年人健康、独立和生存的主要威胁。很多 关于这种下降机制的知识是从针对一种或几种免疫的研究中获得的。 细胞亚群，或批量转录组学。然而，这项工作并没有产生两个关键信息： 1) 对相关免疫细胞群随衰老而发生的集体变化的综合看法，以及 2) 将特定免疫细胞子集与其潜在细胞表型/转录谱联系起来的能力，以及 比较这些表型和概况作为年龄和疫苗反应的函数。单细胞 分析，我们用来涵盖流式细胞术和质谱流式细胞术以及单细胞 RNAseq (scRNAseq) 的术语， 具有独特的优势，可以消除免疫系统异质性并识别新型独特的免疫细胞 健康和疾病的子集。因此，单细胞分析将使我们能够解析免疫细胞子集 与老年人对 PBMC 疫苗的免疫反应相关的缺陷，这是一种高度异质性的起始 细胞群，但具有临床可及性、高度代表性和 最终为了数据生成和分析的目的而公正。我们已经证明我们可以识别 PBMC 的衰老细胞类型特异性免疫特征，即使此类免疫子集代表 PBMC 池总量的一小部分，并拥有来自老年 PBMC 的初步单细胞分析数据， 强调可行性。在这里，我们将在单细胞水平上分析来自老年捐赠者之前和之后的 PBMC。 接种疫苗后（目标 1），以及有或没有体外激活特定免疫子集（目标 2），了解 作为年龄和疫苗的函数而发生或未发生的协调转录和功能变化 反应能力。该提案建立在我们公认的人类免疫学专业知识的基础上，并结合了 细胞计数和单细胞转录组分析 (JAX-GM) 方面的基本专业知识，以及接触老年群体的机会 （乔治·库切尔，康涅狄格大学老龄化中心）。影响：这些研究将以前所未有的分辨率得出 细胞类型特异性免疫特征可区分流感疫苗的反应者和非反应者 为成功疫苗免疫反应的机制和生物标志物提供关键线索。 此外，这些研究将产生大量与以下相关的转录和功能数据： 参与老年人流感疫苗反应的关键先天免疫和 T/B 细胞亚群的输出 个人。这些数据将成为未来研究老年人免疫系统在健康和健康方面的重要资源。 疾病。

PROJECT SUMMARY The goal of this project is to understand the aging-related genomic and functional changes in immune cells that affect responses to flu vaccination. The declining ability of the aging immune system to mount protective responses to vaccines is a major threat to the health, independence and survival of older adults. Much knowledge into the mechanisms of this decline has been gained from studies focused on one or a few immune cell subsets, or on bulk transcriptomics. However, this work has not produced two critical pieces of information: 1) an integrated view of the collective changes across relevant immune cell populations with aging, and 2) the ability to link specific immune cell subsets with their underlying cellular phenotypes/transcriptional profiles, and to compare these phenotypes and profiles as a function of age and responsiveness to vaccines. Single cell profiling, a term we use to encompass flow and mass cytometry together with single cell RNAseq (scRNAseq), is uniquely positioned to deconvolve immune system heterogeneity and identify novel distinct immune cell subsets in health and disease. Single cell profiling will therefore enable us to resolve the immune cell subset deficits relevant to the elderly immune response to vaccines from PBMCs, a highly heterogeneous starting population of cells, but one that offers the advantages of being clinically accessible, highly representative and ultimately unbiased for the purposes of data generation and analysis. We have shown that we can identify discreet cell-type-specific immune signatures of aging from PBMCs, even when such immune subsets represent a small fraction of the total PBMC pool, and have preliminary single cell profiling data from elderly PBMCs, underscoring feasibility. Here, we will analyze PBMCs at the single cell level from elderly donors before and after vaccination (Aim 1), and with or without in vitro activation of specific immune subsets (Aim 2), to understand the coordinated transcriptional and functional changes that occur, or fail to, as a function of age and vaccine responsiveness. This proposal builds on our recognized expertise in human immunology and incorporates essential expertise in cytometry and single cell transcriptomic analysis (JAX-GM), and access to elderly cohorts (George Kuchel, UConn Center on Aging). Impact: These studies will yield, with unprecedented resolution, the cell-type-specific immune signatures that distinguish responders to flu vaccine from non-responders, and will provide critical clues into the mechanisms and biomarkers of a successful vaccine immune response. Furthermore, these studies will generate a considerable amount of transcriptional and functional data related to the outputs of key innate immune and T/B-cell subsets involved in the influenza vaccine response of elderly individuals. The data will be an important resource for future studies of the elderly immune system in health and disease.