Systems investigation of vaccine responses in aging and frailty

流感在21世纪仍然是一个世界性的公共卫生威胁。而流感的发病率 在新冠肺炎大流行期间显著下降，流感肯定会重新出现，并有助于 与SARS-CoV-2一起增加了呼吸道病毒疾病的负担。流感病毒的影响被放大了 由于疫苗提供了不完全的保护，以及新的大流行毒株的间歇性出现。 流感对老年人的影响不成比例，美国90%的死亡病例被归因于 到65岁以上的人患流感；疗养院居民虚弱的高患病率 是特别脆弱的。目前有两种疫苗被批准用于65岁以上的成年人--大剂量 (HD)无佐剂疫苗和MF59佐剂标准剂量(SD)疫苗。在这里，我们将开展 第一项随机研究，比较免疫学、转录学和蛋白质组学特征对这些反应的影响 在养老院的老年居民或在家的初级保健计划中的患者中接种两种疫苗， 与接种相同疫苗的年轻人相比。我们正在继续我们与耶鲁大学的长期合作 老年科，与一组老年医生合作，这些医生担任这些项目的医疗主任 负责每年接种流感疫苗的机构和方案。我们将利用我们的专业知识在 进行人类免疫学研究，特别是那些阐明了先天免疫和基因的研究 疫苗反应的表达特征以及年龄如何影响这些特征；例如，我们携带 对年轻人和老年人对SD疫苗反应的基因表达特征进行了最大规模的研究 连续五个疫苗季节登记在册。在接种疫苗之前和之后获得的样本将 通过CyTOF(核心C)对先天性和获得性免疫细胞的组成、激活进行详细分析 状态和细胞内细胞因子的产生。我们还将对先天免疫PRR进行公正的研究 在外周血单核细胞群体和血小板中的功能。流感血凝素抗体反应的广度 蛋白质将通过对异源病毒株和冷冻电子的微中和研究进行评估 基于显微镜的表位映射。这些调查结果将与已建立的CITE-SEQ WITH CELL相结合 用于单细胞转录分析的散列平台(核心C)，也将与B细胞受体结合 测序以比较年轻人和养老院老年人的反应，并与蛋白质组分析 与波士顿儿童医院HIPC中心合作完成的血清样本。最后，在一个子集中 参赛者，我们将获得接种现场的皮肤活检，以可视化早期细胞浸润和局部 接种疫苗后的淋巴活检以了解组织和继发性免疫反应 使用一种创新的空间转录程序DBiT-Seq.对基因的理解， 流感疫苗的免疫学和蛋白质结构可能识别可能成为靶点的途径 治疗、药物或其他生物治疗，以优化老年人的免疫反应。

Influenza remains a worldwide public health threat in the 21st Century. While the incidence of influenza decreased markedly during the COVID-19 pandemic, influenza is certain to re-emerge and contribute to an increased burden of respiratory viral disease together with SARS-CoV-2. The impact of infliuenza is magnified by a vaccine that affords incomplete protection, and by the episodic appearance of new pandemic strains. Influenza has a disproportionate impact in older adults, with 90% of the deaths in the United States attributed to influenza occurring in individuals over the age of 65; nursing home residents with a high prevalence of frailty are particularly vulnerable. Two vaccines are currently approved for use in adults over age 65—a high-dose (HD) unadjuvanted vaccine and a MF59-adjuvanted standard-dose (SD) vaccine. Here, we will carry out the first randomized study to compare immunologic, transcriptomic and proteomic signatures of response to these two vaccines in older adult residents of nursing homes or patients in a homebound primary care program, compared to young adults receiving the same vaccines. We are continuing our longstanding work with the Yale Section of Geriatrics, collaborating with a group of geriatricians that serve as medical directors for these faciilties and programs who are responsible for annual influenza vaccination. We will leverage our expertise in carrying out studies in human immunology, particularly those that have elucidated innate immune and gene expression signatures of vaccine response and how aging affects these signatures; for example, we carried out the largest study of gene expression signatures of response to SD vaccine in young and older adults enrolled over five consecutive vaccine seasons. Samples obtained prior to and following vaccination will undergo detailed analyses by CyTOF (Core C) for innate and adaptive immune cell composition, activation status, and intracellular cytokine production. We will also carry out unbiased studies of innate immune PRR function in PBMC populations and platelets. The breadth of antibody responses to the influenza hemagglutinin protein will be assessed using microneutralization studies with heterologous viral strains and cryo-electron microscopy-based epitope mapping. These findings will be integrated with an established CITE-seq with cell hashing platform (Core C) for single-cell transcriptomic analyses that will also be combined with B cell receptor sequencing to compare responses in young vs. nursing home older adults, and with proteomic analyses on serum samples done in collaboration with the Boston Children’s Hospital HIPC center. Finally, in a subset of participants, we will obtain skin biopsies of the vaccination site to visualize early cellular infiltrates and regional lymph node biopsies following vaccination to gain insights into immune responses in tissues and secondary lymphoid organs using an innovative spatial transcriptomics program, DBiT-Seq. Understanding the genetic, immunologic and protein architecture of influenza vaccination is likely to identify pathways that could be targets of therapies, drugs or other biological treatments to optimize immune responses in older adults.