An Autoimmune center of excellence for the study of IgG4-related disease

描述(由申请人提供)：在这个项目中，我们将寻求利用多学科方法更好地了解lgG4相关疾病的发病机制。基于这一知识，我们试图影响新疗法的发展，这些疗法不仅适用于这种疾病，而且也适用于其他自身免疫性和纤维性疾病。IgG4相关性疾病是一种多系统疾病，以组织肿胀/S、条带样纤维化、闭塞性静脉炎和血浆IgG4水平升高为特征。虽然这种疾病很可能是自身免疫性的，但不排除对致病微生物或共生微生物的异常反应。在主要的提案中，我们将深入研究lgG4相关疾病受试者中激活的效应器CD4+T细胞克隆。我们将通过下一代测序来检查T细胞受体的谱系，并对这些克隆中的基因和蛋白质表达进行详细分析，以便更好地了解它们是如何产生的，以及它们是如何发挥作用的。我们将通过单细胞rna序列和质量细胞术来了解这些细胞的克隆起源。 效应器T细胞。只有在疾病受试者中才能发现的独特细胞表面蛋白将被识别出来。这些T细胞克隆影响纤维化的机制将被研究。还将鉴定成浆细胞的抗体库，产生与疾病相关的新的人类单抗，并研究克隆扩增的成浆细胞的特性。人类ORFeome文库将与血清和疾病相关的单抗lgG4进行查询，疾病特异性抗原将被鉴定并用作工具来鉴定克隆扩增的CD4+T细胞识别的抗原肽。在合作项目中，将进行研究，以将口腔、组织和肠道微生物群落与遗传易感标记和患者亚群之间的关系联系起来，这些患者具有明显的克隆效应T细胞的功能特征。在试点项目中，将分析活动期疾病受试者中lgG4的糖基化情况，并检测其影响与不同人类Fc受体结合的能力。 相关性：拟议的多学科研究不仅旨在使患有lgG4相关疾病的患者受益，还旨在使许多其他疾病受益，包括许多其他自身免疫性疾病和包括特发性肺纤维化在内的其他疾病。 主要项目：IgG4相关疾病的免疫学研究 项目负责人(PL)：Shiv S.Pillai 描述(申请人提供)：lgG4相关疾病是一种多系统疾病，包括一系列以前描述的综合征，现在都被认为是以肿胀皮损、斑片状纤维化、闭塞性静脉炎和大量 血清IgG4。在许多受试者中，类固醇可以看到临床改善，B细胞枯竭在临床上也是有效的。对这种疾病的发病机制或许多明显无关的疾病中纤维化的分子和细胞基础知之甚少。研究将会 基因表达、蛋白质表达和代谢产物的详细询问将通过全球和单细胞RNAseq、多色流式细胞术、Cytof质谱仪以及克隆扩增的T细胞的液-质联用仪进行，并将进行以了解潜在的治疗途径 对这些T细胞的发育以及它们在这种纤维性炎症性疾病中的效应器功能的意义。LgG4抗体在这种疾病中的可能作用也将被研究。下一代测序以及单细胞克隆和测序策略将用于确定浆细胞的扩张，并建立可能导致疾病的特异性抗体重链-轻链对。因此，将使用人ORFeome表达文库作为抗原源来产生鉴定特定抗原的试剂。B细胞特异性蛋白抗原的测定和重组蛋白的使用将用于辅助T细胞特异性多肽的鉴定。 还将使用基于Fluidigm的MHC II类进行lgG4-RD的遗传易感性研究 基因分型，如果免疫芯片分析表明是外显子组测序。基因和蛋白质表达的全球比较将提示表观遗传学研究的必要性。 相关性：引起严重组织疤痕或纤维化的自身免疫性和炎症性疾病可导致过早死亡，并严重损害幸存者的生活质量。拟议的研究可能不仅有助于lgG4相关疾病的患者，还可能帮助许多其他疾病，包括许多其他自身免疫性疾病和包括特发性肺纤维化在内的其他疾病。

DESCRIPTION (provided by applicant): In this project we will seek to better understand the pathogenesis of lgG4 related disease utilizing a multidisciplinary approach. Based on this knowledge we seek to influence the development of new therapies that may be applicable not just to this disease but to other autoimmune and fibrotic diseases as well. lgG4 related disease is a multisystem disorder characterized by tissue swelling/s, storiform fibrosis, obliterative phlebitis and high levels of plasma lgG4. While this disorder is likely to be autoimmune, an aberrant response to a pathogenic or commensal microbe has not been ruled out. In the principal proposal we will study in depth activated effector CD4+ T cell clones in subjects with lgG4-Related disease. We will examine the T cell receptor repertoire by Next Gen Sequencing, and obtain a detailed analysis of gene and protein expression in these clones in order to better understand how they are generated and how they function. We will perform single cell RNA seq and mass cytometry to understand the clonal origins of these effector T cells. Unique cell surface proteins found only in disease subjects will be identified. The mechanism by which these T cell clones influence fibrosis will be examined. The antibody repertoire of plasmablasts will also be identified, and novel disease related human monoclonal antibodies will be generated and the properties of clonally expanded plasmablasts will also be studied. Human ORFeome libraries will be interrogated with serum and disease related monoclonal lgG4 antibodies and disease specific antigens will be identified and used as tools to identify the antigenic peptides recognized by clonally expanded CD4+ T cells. In the Collaborative project studies will be performed to correlate oral, tissue and gut microbial communities with genetic susceptibility markers and subsets of patients with distinct functional characteristics of the clonal effector T cells in disease subjects. In the Pilot Project the glycosylation of lgG4, in subjects with active disease will be analyzed and its ability to influenc binding to different human Fc receptors will be examined. RELEVANCE: The proposed multidisciplinary studies are designed not only to benefit patients with lgG4-related disease, but to also benefit a host of other disorders including many other autoimmune diseases and other diseases including idiopathic pulmonary fibrosis. Principal Project: Studies on the Immunology of IgG4-related diseases Project Leader (PL): Shiv S. Pillai DESCRIPTION (as provided by applicant): lgG4-Related Disease is a multi-system disorder encompassing a host of previously described syndromes, all now recognized to be characterized by tumescent lesions, storiform fibrosis, obliterative phlebitis and large amounts of serum lgG4. Clinical improvement is seen with steroids in many subjects and B cell depletion is also clinically effective. Very little is known about the pathogenesis of this disorder or about th molecular and cellular basis for fibrosis in a host of apparently unrelated disorders. Studies will be performed to define T cell clonal expansions observed by Next Gen Sequencing approaches in subjects with this disease but with distinct organ involvements. Novel surface markers expressed only on effector T cell clones will be investigated as potential targets for therapy. Detailed interrogation of gene expression protein expression and metabolites will be performed by global as well as single cell RNAseq, multi-color flow cytometry, by Cytof mass cytometry, and liquid chromatography-mass spectrometry on clonally expanded T cells and will be conducted in order to understand pathways of potential therapeutic significance that contributes to the development of these T cells as well as to their effector functions in this fibrotic inflammatory disease. A possible role for lgG4 antibodies in this disordr will also be examined. Next Gen Sequencing as well as single cell cloning and sequencing strategies will be used to define plasmablast expansions and to establish specific antibody heavy-light chain pairs that may contribute to the disease. Reagents will be thus generated to identify specific antigens using human ORFeome expression libraries as source of antigen. Determining the B cell specific protein antigen and the use of recombinant proteins will be used to assist the identification of T cell specific peptides. Studies will also be performed on genetic susceptibility to lgG4-RD using Fluidigm based MHC class II genotyping and if indicated from the Immunochip analyses by Exome sequencing. Global comparisons of gene and protein expression will inform the need for epigenetic profiling studies. RELEVANCE: Autoimmune and inflammatory diseases that cause severe tissue scarring or fibrosis can cause premature death and seriously impair the quality of life for survivors. The proposed studies may help not only patients with lgG4-related disease, but also a host of other disorders including many other autoimmune diseases and other diseases including idiopathic pulmonary fibrosis.