Systems investigation of vaccine responses in aging and frailty

流感仍然是21世纪全球性的公共卫生威胁。虽然流感的发病率 在COVID-19大流行期间，流感明显减少，流感肯定会重新出现，并导致 呼吸道病毒疾病和SARS-CoV-2的负担增加。infliuenza的影响被放大 疫苗提供的保护不完全，以及新的大流行毒株的偶发性出现。 流感对老年人的影响不成比例，美国90%的死亡归因于 65岁以上人群中发生的流感;体弱患病率高的疗养院居民 尤其脆弱。目前有两种疫苗被批准用于65岁以上的成年人， (HD)无佐剂疫苗和MF 59佐剂标准剂量（SD）疫苗。在这里，我们将进行 第一个随机研究，比较免疫学，转录组学和蛋白质组学特征的反应，这些 两种疫苗在养老院的老年居民或在家的初级保健计划中的患者中， 与接受相同疫苗的年轻人相比。我们将继续与耶鲁大学的长期合作 老年病科，与一组老年病学家合作，担任这些机构的医疗主任。 负责每年流感疫苗接种的机构和项目。我们将利用我们的专业知识， 开展人体免疫学研究，特别是阐明先天免疫和基因 疫苗反应的表达特征以及衰老如何影响这些特征;例如，我们进行了 在年轻人和老年人中对SD疫苗反应的基因表达特征的最大研究 在连续五个疫苗接种季节中招募。在接种疫苗之前和之后获得的样品将 通过CyTOF（核心C）对先天性和适应性免疫细胞组成、活化 状态和细胞内细胞因子产生。我们还将开展先天免疫PRR的无偏倚研究 在PBMC群体和血小板中的功能。对流感血凝素的抗体反应宽度 将使用异源病毒株和冷冻电子显微镜进行微量中和研究， 基于显微镜的表位作图。这些研究结果将与已建立的CITE-seq细胞 用于单细胞转录组学分析的散列平台（核心C），也将与B细胞受体结合 测序比较年轻人与养老院老年人的反应，并对 与波士顿儿童医院HIPC中心合作完成的血清样本。最后，在 参与者，我们将获得疫苗接种部位的皮肤活检，以观察早期细胞浸润和区域 接种疫苗后的淋巴结活检，以了解组织和继发性免疫应答 淋巴器官使用创新的空间转录组学程序，DBiT-Seq。了解基因， 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.