Targeted in vivo perturbation for dissecting tissue immune responses

项目3：项目概要 成功的疫苗接种代表了多种相互作用的先天性、体液性和细胞适应性的相互作用 这些因素尚未完全确定。免疫系统作为一个整体参与，包括两个组织 居民和流动人口。虽然我们阐述了综合对策的重要内容， 所选细胞亚型的作用中的许多细微差别仍然有待于单独和组合地确定。 一种解剖免疫反应复杂网络的方法，已成功地应用于模型 系统-和体外人类免疫学-涉及分子特异性，有针对性的干扰基线 细胞相互作用的网络。随后免疫反应的变化可以开始突出免疫反应。 参与疫苗反应的网络元素与靶向的分子实体有关。这里我们 利用多发性硬化症（MS）患者接受分子治疗的事实， 特定的治疗，因此提供了一个机会，在体内检查靶向扰动如何改变 在人类中建立对CMV的应答和对新疫苗的应答。我们关注两种治疗方法 其用作单一疗法：（1）用ocrelizumab去除B细胞和（2）在细胞中隔离淋巴细胞。 用芬戈莫德治疗淋巴结我们建议对接受以下治疗的MS受试者进行纵向抽样： 这两种疗法-和健康受试者作为参考-通过三个疫苗接种周期：两个 SARS-CoV-2疫苗和流感疫苗。在接种前时间点和最多8 随后的时间点，我们将评估体液和T细胞对CMV，流感和SARS-CoV-2的反应， 多种不同的模式，包括高维细胞计数，单细胞CITEseq，和功能 评估对抗原特异性刺激的反应。因此，我们将在一个罕见的位置， 在健康和体内3-4年内对多种不同的抗原特异性刺激的纵向反应 目标扰动上下文。

PROJECT 3: PROJECT SUMMARY Successful vaccination represents the interplay of multiple, interacting innate, humoral, and cellular adaptive factors which have not yet been fully defined. The immune system as a whole is engaged, including both tissue resident and circulating populations. While we have elucidated important elements of the integrated response, many nuances in the role of selected cell subtypes remain to be determined both in isolation and in combination. One approach to dissect complex networks of immune responses that has been applied successfully in model systems - and in vitro for human immunology – involves molecularly specific, targeted perturbation of the baseline network of cellular interactions. Subsequent changes in immune responses can then begin to highlight the elements of the network involved in vaccine response related to the molecular entity that was targeted. Here, we deploy this strategy leveraging the fact that individuals with multiple sclerosis (MS) are treated with molecularly specific treatments and therefore present an opportunity to examine, in vivo, how targeted perturbation alters established responses to CMV and response to new vaccines in humans. We focus on two such treatments which are used as monotherapy: (1) B cell depletion with ocrelizumab and (2) sequestration of lymphocytes in the lymph node using fingolimod. We propose to longitudinally sample participants with MS treated with one of these two therapies - and healthy subjects to serve as a reference – through three vaccination cycles: two with SARS-CoV-2 vaccine and one with an influenza vaccine. At the pre-vaccination time point and up to 8 subsequent time points, we will assess humoral and T cell responses to CMV, influenza and SARS-CoV-2 using multiple different modalities, including high-dimensional cytometric, single cell CITEseq, and functional assessments in response to antigen-specific stimulation. We will therefore be in a rare position to model longitudinal responses to multiple different, antigen-specific stimulations over 3-4 years in the healthy and in vivo targeted perturbation context.