Project 3: Effects of Age on Cellular, Transcriptomic and Biochemical Influenza Signatures

流感仍然是21世纪全球性的公共卫生威胁。它的影响被一种疫苗放大， 提供不完全的保护，其组成通常每年都在变化， 新的大流行毒株，如2009年出现的A/加州H1N1毒株。流感特别 严重影响老年年龄组，美国每年20，000 - 40，000例死亡中有90% 这是由于流感发生在65岁以上的人身上。这个项目，就像所有的项目一样， 人类免疫学项目联盟的应用，旨在确定与免疫相关的宿主因素， 人类的反应。在这里，我们重点关注影响流感疫苗反应的因素，以及这些因素如何影响流感疫苗反应。 因素和途径受年龄和功能状态的影响。在这份提案中，我们将利用我们的专业知识 在进行人类免疫学研究，特别是我们的研究，已确定先天免疫， 疫苗对标准剂量疫苗反应的基因表达特征以及衰老如何影响这些特征 签名.特别是，我们发现了一个意想不到的线粒体生物发生特征， 与疫苗抗体反应相关，这一发现，加上初步的代谢组学数据， 使我们建议使用已经收集血清样品来获得疫苗的生物化学特征 这种反应将与基因表达、细胞和免疫学特征相结合， 在这一群体中，年轻人和老年人都得到了阐明。我们还建议阐明生物学的新见解， 虚弱的基础，一种老年综合征，特征是生理储备和抗应激能力下降， 与死亡率和残疾率的增加有关。我们还将获得新的见解， 最近批准用于老年人的流感疫苗，将该疫苗给予年轻人（21岁- 30岁）、非虚弱老年人和虚弱老年人（≥ 70岁）。对于这些研究，我们将利用大型数据库， 由耶鲁大学老龄化项目建立的成年人和脆弱和脆弱专家的专业知识和跟踪记录， 比如托马斯吉尔博士。接种疫苗前后采集的样本将进行 通过CyTOF（核心C）对先天性和适应性免疫细胞组成的详细分析（包括新的研究 血小板谱系）、活化和细胞因子产生。我们还将进行先天性的无偏见研究， 免疫模式识别受体功能。这些发现将与单细胞RNA-seq 利用核心C中的细胞平台从鉴定为在细胞周期中发生变化的谱系中分离单个细胞的研究 疫苗接种的背景，以及疫苗接种前后血清和细胞的代谢分析。等 代谢组学分析将鉴定反映许多遗传信号输入的整合的途径， 并将为人类流感疫苗反应提供更多的见解。深入了解遗传学、免疫学 流感疫苗的代谢结构可能会识别出可能成为靶点的途径， 治疗、药物或其他生物治疗以根据需要增强或抑制免疫应答。

Influenza remains a worldwide public health threat in the 21st Century. Its impact is magnified by a vaccine that affords incomplete protection whose composition usually changes annually, and by the episodic appearance of new pandemic strains, such as the A/California H1N1 strain that emerged in 2009. Influenza has a particularly acute impact in in the geriatric age group, with 90% of the 20,000-40,000 annual deaths in the United States attributed to influenza occurring in individuals over the age of 65. This proposal, like all of the Projects in this Human Immunology Project Consortium application, seeks to identify the host factors associated with immune response in humans. Here we focus on factors contributing to influenza vaccine response, and how these factors and pathways are affected by age and functional status. In this proposal, we will leverage our expertise in carrying out studies in human immunology, particularly our studies that have identified innate immune and gene expression signatures of vaccine response to standard-dose vaccine and how aging affects these signatures. In particular, we found an unexpected mitochondrial biogenesis signature that was strongly associated with vaccine antibody response, and this finding, together with preliminary metabolomic data, has led us to propose employing already-collected serum samples to derive biochemical signatures of vaccine response that will be integrated with the gene expression, cellular and immunologic signatures already elucidated for young and older adults in this cohort. We also propose to elucidate new insights into the biologic basis of frailty, a geriatric syndrome characterized by decreased physiologic reserve and stress resistance that is associated with increased mortality and disability. We will also gain new insights into the high-dose influenza vaccine recently approved for older adults, administering this vaccine to a cohort of young (age 21- 30), non-frail older and frail older (≥ 70) adults. For these studies, we will leverage large databases of older adults established by the Yale Program on Aging and the expertise and track record of experts in frailty and disability at Yale such as Dr. Thomas Gill. Samples obtained prior to and following vaccination will undergo detailed analyses by CyTOF (Core C) for innate and adaptive immune cell composition (including novel studies of the platelet lineage), activation, and cytokine production. We will also carry out unbiased studies of innate immune pattern recognition receptor function. These findings will be integrated with single-cell RNA-seq studies utilizing the nanowell platform in Core C to isolate individual cells from lineages identified to change in the context of vaccination, and with metabolic analyses of serum and cells pre- and post-vaccine. Such metabolomic analyses will identify pathways reflecting the integration of numerous genetic signaling inputs, and will provide additional insights into human influenza vaccine response. Insights into genetic, immunologic and metabolic architecture of influenza vaccination are likely to identify pathways that could be targets of therapies, drugs or other biological treatments to enhance or suppress immune responses as needed.