University of Chicago Autoimmunity Center of Excellence

芝加哥大学自身免疫卓越中心

• 批准号: 10413988
• 负责人: Marcus Ramsay Clark
• 金额: $ 32.4万
• 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/10413988
• 关键词: Acute; Affect; Affinity; Anatomy; Antibodies; Antigen-Presenting Cells; Architecture; Autoantibodies; Autoantigens; Autoimmune; Autoimmune Diseases; Autoimmunity; B-Lymphocytes; Biological Assay; Blood; CD4 Positive T Lymphocytes; Celiac Disease; Cell Communication; Cell Shape; Cells; Chicago; Dendritic Cells; Development; Disease; Duodenum; Equilibrium; 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; 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 memory cell; 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; macrophage; 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.

项目总结 芝加哥大学ACE(UCACE)的研究重点是原位人类自身免疫。在上一次资助期间 周期，我们成功地开发了完全表征单个B细胞转录状态的技术，并 从组织样本中分离出浆母细胞，并将其与功能免疫球蛋白谱系的分析配对。 此入路在狼疮性肾小管间质炎症(TII)和急性混合移植肾中的应用 排斥反应(AMR)提示抗原驱动的B蛋白的激活状态、抗原库和机制 人类炎症中的细胞选择与通常在继发性炎症中观察到的细胞选择有根本的不同 淋巴器官(合作项目)。在腹腔疾病中，表达转谷氨酰胺酶2(TG2)的B细胞 特异性抗体在十二指肠粘膜中大量扩张。一个高度受限的VH基因谱系 编码这些抗体，最明显的是VH5-51。值得注意的是，这种VH基因在 循环中的IgA+B细胞池，具有丰富的抗微生物活性。这些发现提出了一个模型，其中 微生物抗原在易感宿主中选择致病性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)所独有的。以前的工作有 通过量化多色共焦图像中T细胞和B细胞之间的距离(CELL 距离映射，CDM)，我们可以确定有能力的TFH细胞和B细胞的功能关系。我们 现在已经实现了深度卷积神经网络(DCNN)，可以准确地识别两个细胞 多色共焦图像中的位置和形状。在小鼠中，对DCNN输出(CDM3)的分析表明 T细胞形态与距树突状细胞距离的关系区分同源和非同源树突状细胞 同源T细胞：敏感度和特异度接近双光子发射的DC相互作用 显微镜(TPEM)。在狼疮TII患者中，CDM3均证实髓系树突状细胞呈递CD4+T细胞抗原。 在重症TII中，浆细胞样树突状细胞(DC)是一种重要的抗原提呈细胞。最后，在 在试点项目中，我们正在应用微流控技术开发能够研究同源基因的体外培养系统 单细胞之间的相互作用。这些项目展示了一种新的方法流水线来确定 原位细胞群体，表征它们的功能，并量化它们通过的适应性细胞网络 合作推动局部适应性免疫和炎症。