Studies on the Immunology of IgG4-related diseases

LgG4相关疾病是一种多系统疾病，包括前面描述的一系列综合征， 所有这些现在都被认为是以肿胀的皮损，条带状纤维化，闭塞性静脉炎和 大量血清lgG4。在许多受试者和B细胞中使用类固醇可以看到临床改善 衰竭在临床上也是有效的。关于这种疾病的发病机制或关于 一系列明显无关疾病中纤维化的分子和细胞基础。 将进行研究，以确定通过下一代测序方法观察到的T细胞克隆性扩张 在患有这种疾病但有明显器官受累的受试者中。新的表面标记仅在 效应性T细胞克隆将被作为潜在的治疗靶点进行研究。基因的详细询问 表达蛋白表达和代谢物将由全局和单细胞RNAseq执行， 多色流式细胞术，采用Cytof质谱仪和液-质联用技术 克隆性扩增T细胞，并将进行研究，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个过程中的效应功能的意义 纤维炎症性疾病。 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定义成浆细胞的扩增和建立 可能导致疾病的特异性抗体重链-轻链对。试剂就是这样产生的 以人ORFeome表达文库为抗原源，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细胞特异性多肽。 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。全球范围的比较 基因和蛋白质的表达将使表观遗传学研究成为可能。

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.