Engineered ImmuneChip Platform to Study B cell Migration and Affinity Maturation

项目摘要 开发针对SARS-COV-2和流感等新出现的感染的疫苗需要加强 了解体内潜在的抗体免疫反应。虽然抗体，用作 治疗剂可以来源于融合的杂交瘤模型、动物模型或基因工程，这些 技术不能解释抗体形成的详细免疫学过程，因此，不能 解读或预测宿主-病原体相互作用的结果。对哺乳动物免疫系统的研究 长期局限于体内方法或使用有限供体淋巴组织的单时间点研究， 通常不允许对细胞内和细胞外过程进行多维空间和时间控制， 调节免疫细胞的决定。这是由于淋巴结的复杂性，其具有不同的淋巴结。 B和T细胞、基质细胞和抗原呈递细胞的小生境。当暴露于抗原时，B细胞经历 一个高度受控的激活过程，称为生发中心（GC）反应，它使抗原特异性 抗体分泌细胞。在GC内，幼稚B细胞被激活，增殖，迁移，经历 免疫球蛋白类别转换，并通过体细胞超突变和基于T细胞的 选择，产生对特异性抗原具有高亲和力的长寿命浆细胞。然而，机械 对GC过程的理解主要来自小鼠淋巴结或2D B细胞培养，它们不 生成真实的气相色谱响应。这个R21的目标是开发一个免疫芯片平台，（A） 结合淋巴结的关键分子和细胞成分，以诱导GC反应并使B 细胞迁移（B）通过强制亲和力成熟过程选择高亲和力B细胞。免疫芯片 将提供GC中细胞过程的多维控制，允许快速产生免疫治疗剂， 并作为快速检测平台来鉴定候选疫苗和免疫原。的成功应用 这一项目的实施将有助于快速发现现有和新出现的感染的候选疫苗， 包括致命流感和SARS冠状病毒2。

PROJECT SUMMARY Developing vaccines against emerging infections such as SARS-COV-2 and influenza requires an enhanced understanding of the underlying antibody immune response in the body. Although antibodies, used as therapeutic agents, can be derived from fused hybridoma models, animal models, or genetic engineering, these techniques cannot explain the detailed immunological process of antibody formation and therefore, cannot decipher or predict outcomes of host-pathogen interactions. The study of the mammalian immune system has long been limited to in vivo approaches or single time point studies with limited donor lymphoid tissues, which often do not allow multidimensional spatial and temporal control of intracellular and extracellular processes that regulate the decisions of immune cells. This is attributable to the complexity of lymph nodes, which have distinct niches of B and T cells, stromal cells, and antigen-presenting cells. When exposed to antigens, B cells undergo a highly controlled activation process, called the germinal center (GC) reaction, which makes antigen-specific antibody-secreting cells. Inside GCs, naïve B cells become activated, proliferate, migrate, undergo immunoglobulin class switching, and increase their antigen affinity by somatic hypermutation and T cell-based selection, yielding long-lived plasma cell with high affinity for specific antigens. However, the mechanistic understanding of the GC process is largely derived from mouse lymph nodes or 2D B cell cultures, that do not generate a bona fide GC response. The goal of this R21 is to develop an ImmuneChip platform that (A) incorporates key molecular and cellular components of the lymph nodes to induce GC reactions and enable B cell migration, (B) selects for high-affinity B cells through a forced affinity maturation process. The ImmuneChip will provide multidimensional control of cellular processes in GCs, allow rapid generation of immune therapeutics, and serve as a rapid testing platform to identify candidate vaccines and immunogens. The successful application of this project will facilitate the rapid discovery of vaccine candidates for existing and emerging infections, including lethal influenza and SARS-CoV-2.