Characterization of the Functional Repertoire and Ontogeny of FVIII Humoral Response Across Species: Project 1

项目1 -摘要 因子VIII（FVIII）抑制剂的形成是治疗的主要并发症 血友病A（HA），并与发病率和死亡率增加有关。总目标 该项目的目的是通过了解FVIII是如何 被体液免疫系统锁定。我们假设FVIII特异性体液免疫 免疫应答由多种动态抗体库组成。我们组建了一个 一组来自各种互补科学背景的研究人员， 克服了FVIII免疫原性和B细胞知识方面的主要空白。我们会委聘 创新和最先进的技术，以扩大我们目前的理解的基础 FVIII抑制剂形成机制。我们提出以下目标：目标1：确定 患者中循环抗FVIII抗体的克隆库和表位特异性 在HA。采用一种新的免疫组学方法，我们将首次描述 详细描述了由FVIII特异性抗体识别的FVIII表位以及 HA患者相关B细胞与抑制物的关系。目标2：定义FVIII 抗体库及其在犬HA抑制剂模型中的动力学。利用抗原 首次在犬疾病模型中进行噬菌体展示，我们将纵向表征 FVIII驱动的体液免疫反应，这将提供对内在免疫反应的深入了解。 FVIII的免疫原性。这项工作的结果将与目标1至 进一步验证了HA犬模型。目的3：探讨B细胞存活因子的作用 在抑制剂形成、表位多样性和根除方面。新的证据表明B 细胞存活细胞因子，包括BLyS，在免疫介导的疾病。我们的初步数据 表明BLyS可能在FVIII抑制剂中发挥作用。在此，我们将全面 通过一系列人体研究评价BLyS对FVIII免疫原性的贡献 和鼠HA模型。这些研究将共同提供关于体液免疫的见解， 系统，包括建立新的治疗框架， 方法和测试小型和大型临床前模型的有效性。

Project 1 - Abstract The formation of Factor VIII (FVIII) inhibitor is a main complication of the treatment of hemophilia A (HA) and it is associated with increased morbidity and mortality. The overall goal of this project is to gain insights into why FVIII is immunogenic by understanding how FVIII is targeted by the humoral immune system. We hypothesize that the FVIII-specific humoral immune response is composed of a diverse and dynamic antibody repertoire. We assembled a group of investigators from a variety of complementary scientific background that together will overcome major gaps in knowledge on FVIII immunogenicity and B cells. We will employ innovative and state-of-art technologies to broad our current understanding of the underlying mechanism of inhibitor formation to FVIII. We propose the following aims: Aim 1: Define the clonal repertoire and epitope specificity of circulating anti-FVIII antibodies in patients with HA. Taking a novel immunomics approach, we will, for the first time, characterize in detail the FVIII epitopes recognized by FVIII-specific antibodies as well as clonal relationships of the related B cells in HA patients with inhibitors. Aim 2: Define the FVIII antibody repertoire and its dynamics in canine HA inhibitor models. Utilizing antigen phage display for the first time in canine disease models, we will longitudinally characterize the FVIII-driven humoral immune response, which will provide insight into the intrinsic immunogenicity of FVIII. Results from this work will be compared to data from Aim 1 to further validate the HA dog model. Aim 3: Investigate the role of B cell survival cytokines in inhibitor formation, epitope diversity and eradication. Emerging evidence implicates B cell survival cytokines, including BLyS, in immune-mediated diseases. Our preliminary data suggests that BLyS may have a role in FVIII inhibitors. Herein, we will comprehensively evaluate the contribution of BLyS to FVIII immunogenicity through a series of studies in human and murine HA models. Together these studies will provide insights on the humoral immune system in FVIII immunogenicity, including establishing a framework for novel therapeutic approaches and testing the validity of small and large preclinical models.