University of Chicago Autoimmunity Center of Excellence

芝加哥大学自身免疫卓越中心

• 批准号: 9920098
• 负责人: Marcus Ramsay Clark
• 金额: $ 44.4万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-10 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9920098
• 关键词: Acute; Affect; Affinity; Anatomy; Antibodies; Antigen-Presenting Cells; Architecture; Autoantibodies; Autoantigens; Autoimmune Diseases; Autoimmune Process; Autoimmunity; B-Lymphocytes; Biological Assay; Blood; CD4 Positive T Lymphocytes; Celiac Disease; Cell Communication; Cell Shape; Cells; Chicago; Dendritic Cells; Development; Disease; Duodenum; Flow Cytometry; Funding; Genes; Genetic Transcription; Gluten; Helper-Inducer T-Lymphocyte; Human; Immune; Immune response; Immune system; Immunity; Immunoglobulin A; Immunoglobulins; In Situ; In Vitro; Individual; Inflammation; Inflammatory; Kidney; Link; Lupus; Lupus Nephritis; Mediating; Memory; Methodology; Methods; Microfluidics; Microscopy; Modeling; Morbidity - disease rate; Mucous Membrane; Mus; Myelogenous; Organ; Output; Pathogenicity; Pilot Projects; Plasmablast; Population; Positioning Attribute; Process; Rheumatoid Arthritis; Sampling; Sensitivity and Specificity; Shapes; Site; Specificity; System; T-Lymphocyte; Techniques; Testing; Tissue Sample; Tissues; Universities; Work; adaptive immunity; allograft rejection; antimicrobial; base; confocal imaging; convolutional neural network; disease heterogeneity; high throughput analysis; innovation; insight; microorganism antigen; molecular phenotype; mortality; novel; peripheral blood; response; secondary lymphoid organ; single-cell RNA sequencing; transglutaminase 2; two-photon

PROJECT SUMMARY The focus of the University of Chicago ACE (UCACE) is in situ human autoimmunity. During the last funding cycle, we successfully developed techniques to fully characterize the transcriptional state of single B cells and plasmablasts sorted from tissue samples and to pair this with analyses of functional immunoglobulin repertoire. Application of this approach to lupus tubulointerstitial inflammation (TII) and renal acute mixed allograft rejection (AMR) suggest that the activation state, antigenic repertoire and mechanisms of antigenic-driven B cell selection in human inflammation is fundamentally different than that typically observed in secondary lymphoid organs (Collaborative Project). In Celiac disease, B cells expressing transglutaminase 2 (TG2) specific antibodies are massively expanded in the duodenal mucosa. A highly-restricted repertoire of VH genes encode these antibodies, most notably VH5-51. Remarkably, this VH gene is also over-represented in the recirculating IgA+ B cell pool which is rich in anti-microbial activity. These findings suggest a model in which microbial antigens select for pathogenic TG2 reactive antibodies in susceptible hosts (Principle Project). Our second technical innovation is unique to the UCACE (Collaborative Project). Previous work has demonstrated that by quantifying the distance between T and B cells in multicolor confocal images (Cell Distance Mapping, CDM) we could identify competent TFH cells and functional relationships with B cells. We have now implemented a deep convolutional neural network (DCNN) that accurately identifies both cell position and shape in multicolor confocal images. In mice, analysis of the DCNN output (CDM3) indicates that T cell shape as a function of distance from dendritic cells (DCs) discriminates between cognate and non- cognate T cell:DC interactions with a sensitivity and specificity approaching that of two-photon emission microscopy (TPEM). In lupus TII, CDM3 both confirmed that myeloid DCs present antigen to CD4+ T cells in situ and identified plasmacytoid DCs as an important antigen presenting cell (APC) in severe TII. Finally, in the Pilot Project, we are applying microfluidics to develop in vitro culture systems capable of studying cognate interactions between single cells. These projects demonstrate a novel pipeline of methodologies to identify in situ cell populations, characterize their function and quantify the adaptive cell networks through which they cooperate to drive local adaptive immunity and inflammation.

项目摘要 芝加哥大学ACE（UCACE）的重点是原位人类自身免疫。在最后一次融资中 周期，我们成功地开发了技术，以充分表征单个B细胞的转录状态， 从组织样品中分选浆母细胞，并将其与功能性免疫球蛋白库的分析配对。 该入路在狼疮性肾小管间质炎和急性混合移植肾中的应用 排斥反应（AMR）的研究表明，抗原驱动的B的激活状态，抗原库和机制， 人类炎症中的细胞选择与在继发性炎症中通常观察到的细胞选择是根本不同的。 淋巴器官（合作项目）。在乳糜泻中，表达转氨酶2（TG 2）的B细胞 特异性抗体在十二指肠粘膜中大量扩增。高度限制的VH基因库 编码这些抗体，特别是VH 5 -51。值得注意的是，这种VH基因在人类中也过度表达。 富含抗微生物活性的再循环伊加+ B细胞池。这些发现提出了一个模型，其中 微生物抗原选择易感宿主中的致病性TG 2反应性抗体（原理项目）。 我们的第二项技术创新是UCACE（合作项目）独有的。先前的工作已经 证明了通过定量在荧光共聚焦图像中T和B细胞之间的距离（Cell 距离作图，CDM），我们可以鉴定感受态TFH细胞和与B细胞的功能关系。我们 现在，我已经实现了一个深度卷积神经网络（DCNN），可以准确地识别两个细胞 位置和形状的共焦图像。在小鼠中，对DCNN输出（CDM 3）的分析表明， T细胞形状作为与树突状细胞（DC）的距离的函数区分同源和非同源T细胞。 同源T细胞：DC相互作用的灵敏度和特异性接近双光子发射 显微镜（TPEM）。在TII型狼疮中，CDM 3均证实髓样DCs向TII型狼疮中的CD 4 + T细胞呈递抗原。 原位和确定浆细胞样DC作为一个重要的抗原呈递细胞（APC）在严重的TII。最后在 试点项目，我们正在应用微流体技术开发能够研究同源的体外培养系统， 单细胞之间的相互作用。这些项目展示了一种新的方法学管道， 原位细胞群体，表征其功能并量化适应性细胞网络， 协同驱动局部适应性免疫和炎症。