Identification of Metabolic and Immune Deficits in the Aged Population and Their Restoration to Achieve Youthful Anti-Influenza Vaccine Responsiveness

项目摘要 流感是老年人死亡的主要原因，目前的疫苗只保护了这一人群的一小部分 尽管有广泛的疫苗接种计划和专门的配方。更好地理解分子 迫切需要控制免疫衰老的机制，以便新设计的疫苗、佐剂 可以使用药物干预来恢复老年人的年轻抗体(Ab)反应 人口。我们的首要目标是确定衰老对免疫细胞的影响。 抗流感抗体疫苗的反应。我们将全面、全面地评估 外周血中T滤泡辅助细胞(TFH)、T滤泡调节细胞(TFR)和B细胞及其亚群 在季节性流感疫苗接种前后的年轻人和老年人。我们将检验一个假设，即 增龄的结果是改变了TFH/TFR细胞亚群的分布，导致 代谢重新编程和缺陷的B细胞引起流感疫苗反应。我们的综合分析 包括在机体和细胞水平上调查代谢途径中与年龄相关的下降的研究， 例如线粒体生物发生缺陷和一种碳代谢，并确定这些变化 导致免疫功能障碍。最后，从这些研究中获得的知识将用于 概念验证拯救实验在体外恢复最佳的流感疫苗反应性。支持 这些研究的样本将来自153名参与者的季节性流感疫苗接种队列(21-76岁， 包括15名在四价血凝后0、7和32天献血的人(≥65年) (Ha)接种疫苗。我们计划有针对性地扩大这一群体，以招募额外的百名捐赠者≥65年 年龄和一组25名较年轻的成年人(≤40岁，n=25)进行单细胞基因组和代谢组学研究，需要 新鲜的血样。我们将暂时跟踪流感疫苗的反应，并比较年轻人和老年人 组。具体地说，在目标1中，我们将检查子集组成和代谢重新编程的变化 在青年和老年人的滤泡T细胞中。这些研究的完成将使我们能够了解 与衰老相关的体液免疫缺陷是否由TFH和TFR亚群的变化所介导， 内在功能，或两者兼而有之。在目标2中，我们将评估抗体反应的特征并确定 青年和老年人B细胞群的组成和代谢变化。在完成时 通过这些研究，我们还将了解导致免疫印记的变化。在目标3中，我们将分析年龄- 流感疫苗抗原对体外培养的TfH、TfR和B细胞相关功能的影响TFH/B细胞共培养 将接受免疫调节剂和小分子治疗，以恢复免疫和代谢功能 克服疫苗反应中与年龄有关的缺陷。我们已经组建了一支非常有成就的团队 开展这些研究的目的是测试新的范式，并建立一个可行的路线图，说明如何 提高老年人对疫苗的反应性。

Project Summary Influenza is a leading cause of death in the elderly and current vaccines only protect a fraction of this population despite widespread vaccination programs and specialized formulations. A better understanding of the molecular mechanisms that control immunological aging is urgently needed so that newly designed vaccines, adjuvants and pharmacologic interventions can be employed to restore youthful antibody (Ab) responses in the older population. Our overarching goals are focused on determining the effects of aging on immune cells that mediate the anti-influenza Ab vaccine response. We will perform an integrated and comprehensive evaluation of circulating T follicular helper (Tfh), T follicular regulatory (Tfr) cells and B cells as well as their subsets from young and older individuals prior to and after seasonal influenza vaccination. We will test the hypothesis that a consequence of advancing age is an altered distribution of individual subsets of Tfh/Tfr cells resulting in metabolic reprogramming and defective B cell elicited influenza vaccine responses. Our integrated analysis includes studies to investigate at the organismal and cellular level age-related declines in metabolic pathways, such as defective mitochondrial biogenesis and one carbon metabolism and to determine if these changes contribute to the immune dysfunction. Finally, knowledge gained from these studies will be used to perform proof of concept rescue experiments in vitro to restore optimal influenza vaccine responsiveness. To support these studies, samples will be used from a seasonal influenza vaccinee cohort of 153 participants (ages 21-76, including 15 individuals ≥ 65yr) who have donated blood at days 0, 7 and 32 after quadrivalent hemagglutinin (HA) vaccination. We plan a targeted expansion of this cohort to recruit an additional 100 donors ≥ 65 years of age and a group of 25 younger adults (≤ 40yr, n=25) for single cell genomic and metabolomic studies that require fresh blood samples. We will temporally follow the influenza vaccine response and compare young and older groups. Specifically, in Aim 1 we will examine alterations in subset composition and metabolic reprogramming in follicular T cells in young and aged individuals. The completion of these studies will allow us to understand whether defects in humoral immunity associated with aging are mediated by changes in Tfh and Tfr subsets, intrinsic functionality, or both. In Aim 2 we will assess characteristics of the Ab response and determine composition and metabolic changes in B cell populations in young and aged individuals. At the completion of these studies we will also understand changes that result in immune imprinting. In Aim 3 we will analyze age- related dysfunction of Tfh, Tfr and B cells in vitro in response to influenza vaccine antigens. Tfh/B cell co-cultures will be treated with immune modulators and small molecules to restore immunologic and metabolic function to overcome age-related defects in vaccine responses. We have assembled a highly accomplished team to carry out these studies with the goal of testing new paradigms and establishing an actionable roadmap on how to improve vaccine responsiveness in the elderly.