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

项目总结 这个项目的目标是了解衰老相关的基因组和免疫功能的变化。 影响流感疫苗接种反应的细胞。老化的免疫系统装载能力的下降 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。大有可为 通过对一种或几种免疫的研究，已经获得了关于这种下降的机制的知识 细胞亚群，或大量转录本。然而，这项工作并没有产生两条关键的信息： 1)跨相关免疫细胞群体随年龄的集体变化的综合视图，以及2) 能够将特定的免疫细胞亚群与其潜在的细胞表型/转录图谱联系起来，以及 比较这些表型和特征随年龄和对疫苗的反应而变化。单细胞 图谱，我们用来包括流式细胞术和质量细胞术以及单细胞RNAseq(ScRNAseq)的术语， 在消除免疫系统异质性和识别新的独特免疫细胞方面具有独特的地位 健康和疾病中的子集。因此，单细胞图谱将使我们能够解析免疫细胞亚群 老年人对外周血单核细胞疫苗的免疫反应缺陷，这是一个高度异质性的开始 细胞群体，但提供的优势是临床可获得的，高度代表性和 最终为数据生成和分析的目的不偏不倚。我们已经证明，我们可以识别 来自PBMC的谨慎的细胞类型特异的衰老免疫特征，即使这样的免疫亚群代表 只占整个PBMC池的一小部分，并拥有来自老年PBMC的初步单细胞图谱数据， 强调可行性。在这里，我们将在单细胞水平上分析老年捐赠者之前和 在接种疫苗后(目标1)，以及体外激活或不激活特定免疫亚群(目标2)，以了解 作为年龄和疫苗的函数而发生或不发生的协调的转录和功能变化 响应性。这项建议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 在细胞学和单细胞转录分析(JAX-GM)以及接触老年人群方面的基本专业知识 (乔治·库切尔，康涅狄格州老年中心)。影响：这些研究将以前所未有的决心产生 区分流感疫苗应答者和无应答者的细胞类型特异性免疫特征，并将 为成功的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 老年人流感疫苗免疫应答中关键先天免疫和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健康和免疫系统的重要资源 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.