Studies on the Immunology of IgG4-related diseases

IgG 4相关疾病是一种多系统疾病，包括许多先前描述的综合征， 现在认为所有这些都以肿胀性病变、Storiform纤维化、闭塞性静脉炎和 大量的血清IgG 4。在许多受试者和B细胞中观察到类固醇的临床改善 消耗在临床上也是有效的。关于这种疾病的发病机制或关于这种疾病的发病机制， 分子和细胞的基础上纤维化的主机显然无关的疾病。 将进行研究以确定通过下一代测序方法观察到的T细胞克隆扩增 在患有这种疾病但有不同器官受累的受试者中。新的表面标志物仅在 将研究效应T细胞克隆作为治疗的潜在靶。详细询问基因 表达蛋白表达和代谢物将通过全局以及单细胞RNAseq进行， 多色流式细胞术、流式细胞仪、质谱仪和液相色谱-质谱联用技术 克隆扩增的T细胞，并将进行，以了解潜在的治疗途径， 这些T细胞的发育以及它们在这一过程中的效应子功能的重要性， 纤维化炎性疾病 还将检查IgG 4抗体在该病症中的可能作用。下一代测序以及 单细胞克隆和测序策略将用于定义浆母细胞扩增并建立 特异性抗体重-轻链对可能导致疾病。因此将生成试剂 使用人ORFeome表达文库作为抗原来源鉴定特异性抗原。确定 B细胞特异性蛋白抗原和重组蛋白的使用将用于辅助鉴定 T细胞特异性肽。 还将使用基于Fluidigm的MHC II类对IgG 4-RD的遗传易感性进行研究 基因分型，并且如果免疫芯片分析指示，则通过外显子组测序。全球比较 基因和蛋白质表达将告知表观遗传分析研究的需要。

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 the 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 contribute 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 disorder 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.