Intrarenal B cells in acute kidney allograft rejection

肾内 B 细胞在急性同种肾移植排斥反应中的作用

• 批准号: 10329990
• 负责人: Anita S Chong
• 金额: $ 79.13万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10329990
• 关键词: Ablation; Acute; Address; Allogenic; Antibodies; Antibody Specificity; Antigen Presentation; Antigen-Presenting Cells; Antigenic Specificity; Antigens; Architecture; Autoimmune Diseases; Automobile Driving; B-Cell Antigen Receptor; B-Lymphocytes; Biological Markers; Biopsy; Blood; Cell Communication; Cell Shape; Cells; Characteristics; Clinic; Clinical; Collaborations; Complement; Complex; Coupled; Dendritic Cells; Ensure; Genetic Transcription; Glucocorticoids; Graft Rejection; Helper-Inducer T-Lymphocyte; Heterogeneity; Human; Immune; Immunity; Immunoglobulin Variable Region; Immunosuppression; In Situ; Inflammation; Inflammatory; Investigation; Kidney; Kidney Transplantation; Measures; Mediating; Methods; Monoclonal Antibodies; Mus; Outcome; Output; Paper; Pathogenicity; Peripheral; Phenotype; Play; Population; Positioning Attribute; Process; Property; Reporting; Resistance; Role; Seminal; Sensitivity and Specificity; Shapes; Specificity; Steroid Resistance; T-Cell Activation; T-Lymphocyte; T-Lymphocyte Subsets; Techniques; Testing; Therapeutic Intervention; Tissue Sample; adaptive immunity; allograft rejection; antibody-mediated rejection; cell type; convolutional neural network; donor-specific antibody; effector T cell; experimental study; genetic signature; high throughput technology; human disease; human tissue; immune function; innovation; kidney allograft; kidney biopsy; molecular targeted therapies; mouse model; neural network; new therapeutic target; novel; novel strategies; peripheral blood; programmed cell death protein 1; response; secondary lymphoid organ; single-cell RNA sequencing; standard of care; therapeutically effective; transplant model; two photon microscopy

ABSTRACT In a seminal paper, Sarwal et al.1 reported that the presence of a B cell gene signature and dense clusters of B cells in the renal biopsies was strongly associated with glucocorticoid-resistant T cell-mediated graft rejection (TCMR) and kidney allograft loss. They proposed a hypothesis that, despite standard of care immunosuppression, infiltrating B cells play a pivotal role in a subset of acute cellular rejection of kidney allografts in the clinic. While some subsequent studies supported this hypothesis, others reported opposite outcomes. As a result, the role of intrarenal B cells in clinical TCMR is currently unclear, and an understanding of their role is urgently required to allow clinicians to rationally decide if B cell depletion might be useful for reversing steroid-resistant TCMR. We hypothesize that the identification of additional features of intrarenal B cells or of the rejecting biopsy are required to more accurately predict poor graft outcome and the need for B cell depletion to treat TCMR. These features include an understanding the specificity of intrarenal B cells and their functional properties, as well as the inflammation architecture of the biopsy. To this end, we have developed two innovative approaches to understand in situ adaptive T and B cell immunity in human biopsies from rejecting kidney allografts. First, we are able to pair the transcriptional state in single B cells with the antigenic specificity of the antibody they secrete. Sorted B cells from renal biopsies are subjected to single cell (sc) RNA-Seq, and the immunoglobulin variable regions are cloned from these same cells and expressed corresponding antibodies. Second, we have developed a novel approach to characterize the spatial architecture between different immune cell populations in human tissue and identify functional relationships2,3. Using a novel deep convolutional neural network (DCNN), coupled to a tuned neural network (TNN), we can accurately capture T cell shape change upon recognition of antigen presented by dendritic cells (DCs) in multicolor confocal micrographs. Therefore, this analytic pipeline (Cell Distance Mapping, CDM) can identify and quantify antigen presentation in fixed tissue samples from both mice and humans. We will use CDM to study T cell interactions with B cells or dendritic cells (DCs) in rejecting kidney biopsies. Third, we developed a mouse model of kidney transplantation to address mechanistic questions raised by observations made from the human renal biopsies. We will apply these three approaches to test our project hypothesis in two Aims: Aim 1. Test the hypothesis that the different transcriptional and functional states of intrarenal B cells during rejection are associated with B cell receptor (BCR) specificity and rejection type. Aim 2: Test the hypothesis that the type of cellular rejection (T cell mediated rejection vs. mixed T cell- and antibody-mediated rejection) is defined by characteristic in situ cellular interactions between T cells and B cells or DCs that are amenable to specific therapeutic intervention.

抽象的 在一篇开创性论文中，Sarwal 等人 1 报道了 B 细胞基因特征和密集簇的存在 肾活检中 B 细胞的减少与糖皮质激素耐药性 T 细胞介导的移植物密切相关 排斥反应（TCMR）和肾同种异体移植物丢失。他们提出了一个假设，尽管护理标准 免疫抑制、浸润 B 细胞在肾急性细胞排斥反应中发挥关键作用 在诊所进行同种异体移植。虽然随后的一些研究支持了这一假设，但其他研究却得出了相反的结论 结果。因此，肾内 B 细胞在临床 TCMR 中的作用目前尚不清楚，并且需要了解 迫切需要了解它们的作用，以便临床医生能够理性地决定 B 细胞耗竭是否对以下疾病有用： 逆转类固醇耐药 TCMR。我们假设肾内 B 的其他特征的识别 需要细胞或排斥性活检来更准确地预测不良的移植结果和 B 的需要 细胞耗竭来治疗 TCMR。这些特征包括了解肾内 B 细胞的特异性和 它们的功能特性，以及活检的炎症结构。为此，我们有 开发了两种创新方法来了解人体活检中的原位适应性 T 和 B 细胞免疫 因排斥同种异体肾移植物。首先，我们能够将单个 B 细胞中的转录状态与 它们分泌的抗体具有抗原特异性。对来自肾活检的分选 B 细胞进行单细胞分析 (sc) RNA-Seq，免疫球蛋白可变区从这些相同的细胞中克隆并表达 相应的抗体。其次，我们开发了一种新颖的方法来表征空间 人体组织中不同免疫细胞群之间的结构并确定功能关系2,3。 使用新颖的深度卷积神经网络（DCNN）与调谐神经网络（TNN）相结合，我们可以 在识别树突状细胞 (DC) 呈递的抗原后，准确捕获 T 细胞形状变化 多色共焦显微照片。因此，这个分析管道（小区距离映射，CDM）可以识别 并量化小鼠和人类固定组织样本中的抗原呈递。我们将使用CDM 研究排斥肾活检时 T 细胞与 B 细胞或树突状细胞 (DC) 的相互作用。第三，我们开发了一个 肾移植小鼠模型，以解决由观察结果提出的机制问题 人类肾活检。我们将应用这三种方法来测试我们的项目假设的两个目标： 目的 1. 检验以下假设：肾内 B 细胞在 排斥反应与 B 细胞受体 (BCR) 特异性和排斥类型有关。 目标 2：检验细胞排斥类型（T 细胞介导的排斥与混合 T 细胞介导的排斥）的假设 抗体介导的排斥）是由 T 细胞和 B 细胞之间的原位细胞相互作用特征定义的 或适合特定治疗干预的 DC。