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 Sarwal等人在开创性纸中报告说，B细胞基因的存在和致密簇的存在 肾脏活检中的B细胞与糖皮质激素耐药的T细胞介导的移植物密切相关 排斥（TCMR）和肾脏同种异体移植损失。他们提出了一个假设，尽管标准了 免疫抑制，浸润B细胞在肾脏急性细胞排斥的子集中起关键作用 诊所中的同种异体移植。虽然随后的一些研究支持了这一假设，但其他研究报告相反 结果。结果，目前尚不清楚肾内B细胞在临床TCMR中的作用，并且理解 迫切需要迫切需要其作用，以便允许临床医生合理决定B细胞耗竭是否可能对 逆转抗类固醇的TCMR。我们假设识别肾内B的其他特征 细胞或拒绝活检需要更准确地预测较差的移植结果，并且需要B 细胞耗竭以治疗TCMR。这些功能包括了解肾内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 开发了两种创新的方法来理解人类活检中的原位适应性T和B细胞免疫力 拒绝肾脏同种异体移植。首先，我们能够将单个B细胞中的转录状态与 它们分泌的抗体的抗原特异性。肾脏活检的分类B细胞受到单细胞的约束 （SC）RNA-SEQ和免疫球蛋白变量区域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体之间的结构并确定功能关系2,3。 使用新型的深卷积神经网络（DCNN），结合到调谐神经网络（TNN），我们可以 在识别树突状细胞（DC）中识别抗原时，准确捕获T细胞形状的变化 多色共聚焦显微照片。因此，该分析管道（单元距离映射，CDM）可以识别 并量化来自小​​鼠和人类的固定组织样品中的抗原表现。我们将使用CDM到 研究T细胞与B细胞或树突状细胞（DC）的T细胞相互作用在拒绝肾脏活检中。第三，我们开发了 肾脏移植的鼠标模型，以解决由由 人类的肾脏活检。我们将应用这三种方法以两个目的测试我们的项目假设： AIM 1。检验以下假设：在 排斥与B细胞受体（BCR）特异性和排斥类型有关。 目标2：检验细胞排斥类型的假设（T细胞介导的排斥反应与混合T细胞和混合T细胞和 抗体介导的排斥反应）由T细胞与B细胞之间的原位点细胞相互作用定义 或适合特定治疗干预的DC。