Targeted in vivo perturbation for dissecting tissue immune responses

项目3：项目总结 成功的疫苗接种代表了多种、相互作用的先天、体液和细胞适应性的相互作用。 尚未完全定义的因素。免疫系统作为一个整体参与，包括这两个组织 常住人口和流动人口。虽然我们已经阐明了综合反应的重要要素， 所选细胞亚型作用的许多细微差别仍有待单独和组合确定。 一种剖析免疫反应复杂网络的方法，已在模型中成功应用 系统--以及体外人类免疫学--涉及到对基线的分子特异性、有针对性的扰动 蜂窝交互网络。随后免疫反应的变化可以开始突显 参与疫苗反应的网络元件与目标分子实体相关。在这里，我们 利用多发性硬化症(MS)患者可以通过分子疗法治疗的事实，部署这一战略 特定的治疗方法，因此提供了一个在体内检查靶向干扰如何改变的机会 人类对巨细胞病毒和新疫苗的已有反应。我们专注于两种这样的治疗方法 它们被用作单一疗法：(1)用奥曲珠单抗去除B细胞和(2)隔离淋巴细胞。 用指状物检测淋巴组织。我们建议对患有多发性硬化症的患者进行纵向抽样，接受以下治疗之一 这两种疗法--并以健康受试者为参照--通过三个疫苗周期：两个 一种是SARS-CoV-2疫苗，一种是流感疫苗。在接种前的时间点和最多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，包括高维细胞术、单细胞CITEseq和功能性 对抗原特异性刺激的反应评估。因此，我们将处于一个难得的位置来建模 3-4年健康人和活体人对多种不同抗原特异性刺激的纵向反应 有针对性的扰动背景。

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.