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 芝加哥大学王牌（UCACE）的重点是原位人类自身免疫性。在上次资金中 循环，我们成功地开发了技术以充分表征单个B细胞的转录状态和 从组织样品分类的浆膜，并将其与功能性免疫球蛋白库的分析配对。 将这种方法应用于狼疮性微管炎症（TII）和肾脏急性混合同种异体移植物 排斥（AMR）表明激活状态，抗原库和抗原驱动的B的机制 人类炎症中的细胞选择与次级中通常观察到的细胞炎症基本不同 淋巴器官（协作项目）。在腹腔疾病中，B细胞表达转谷氨酰胺酶2（TG2） 在十二指肠粘膜中大量扩展了特定的抗体。 VH基因的高度限制曲目 编码这些抗体，最著名的是VH5-51。值得注意的是，该VH基因在 富含抗微生物活性的IgA+ B细胞池。这些发现提出了一个模型 微生物抗原选择易感宿主中的致病性TG2反应性抗体（原理项目）。 我们的第二项技术创新是Ucace（协作项目）独有的。以前的工作有 证明，通过量化多色共聚焦图像中T和B细胞之间的距离（细胞 距离映射，CDM）我们可以鉴定有效的TFH细胞和与B细胞的功能关系。我们 现在已经实施了一个深度卷积神经网络（DCNN），该神经网络准确地识别了两个细胞 多色共聚焦图像中的位置和形状。在小鼠中，对DCNN输出的分析（CDM3）表明 T细胞形状与距离树突细胞距离（DC）的函数，可区分cognate和non- 同源T细胞：DC相互作用与敏感性和特异性接近两光子发射 显微镜（TPEM）。在狼疮TII中，CDM3都证实髓样DC在CD4+ T细胞中呈现抗原 原位并确定质晶型DC是严重TII中重要的抗原呈现细胞（APC）。最后，在 试点项目，我们正在应用微流体来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颖的方法，可以识别 原位细胞种群，表征其功能并量化它们的自适应细胞网络 合作驱动局部适应性免疫和炎症。