Human B cell infiltration into allografts: mechanisms and molecules

人 B 细胞浸润同种异体移植物：机制和分子

基本信息

批准号：
10043269
负责人：
Thomas David Manes
金额：
$ 25.13万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-21 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10043269
关键词：
Ablation Acute Adhesives Alloantigen Allografting Animal Model Antibodies Antigen Presentation Antigens Archives B-Lymphocytes Blood Circulation Blood Vessels CD3 Antigens Cell Adhesion Molecules Cell physiology Chronic Clinical Trials Cloning Complement Data Endothelial Cells Event Goals Human Immune Immune system Immunocompromised Host Immunofluorescence Immunologic Immunofluorescence Microscopy Immunologics Immunosuppression In Vitro Individual Infiltration Intervention Kidney Transplantation Lasers Light Mediating Memory Microfluidics Microscopy Migration Assay Molecular Target Mus Organ Transplantation Pathogenesis Pathway interactions Pattern Periodicity Pharmaceutical Preparations Plasma Cells Plasmablast Population Process Receptor Signaling Receptors, Antigen, B-Cell Risk Sampling Signal Transduction Source Specificity T cell response T memory cell T-Cell Receptor T-Lymphocyte Techniques Testing Therapeutic Tissues Work adhesion process adhesion receptor cell type chemokine chemokine receptor crosslink cytokine effective therapy high dimensionality human migration in vitro Assay in vitro Model insight kappa opioid receptors kidney allograft migration monolayer novel therapeutic intervention outcome forecast peripheral blood prevent receptor recruit response tertiary lymphoid organ

项目摘要

7. PROJECT SUMMARY/ABSTRACT Rates of late renal allograft loss due to chronic rejection have not significantly changed over the past three decades despite marked improvements in controlling acute rejection. While the immune pathogenesis is incompletely understood, a significant proportion of allografts contain infiltrated host B cells and the presence of B cells correlates with poor prognosis that may be independent of the presence of a donor specific antibody. While T cell recruitment to allografts is well understood and may be initiated by either chemokines or antigen presentation within the vascular lumen, much less is known about B cell recruitment. We propose to study these processes with the goal of identifying pathways and molecular targets that may inhibit B cell infiltration of grafts. To study if and how this process can be induced by chemokines, we will employ both high dimensional cyclic tissue immunofluorescence microscopy of human allograft tissue and in vitro assays of transendothelial migration (TEM) by human peripheral blood B cells. Tissue analyses will provide information re the subtypes of B cells found within graft infiltrates, the chemokine and adhesion receptors they express, and their proximity to activated endothelial cells that are the likely point of entry. In vitro assays will use human microvascular endothelial cell monolayers in microfluidic chambers to characterize and manipulate B cells capable of undergoing TEM using approaches we developed in our T cell studies. To study if and how this process can be induced by antigen, we will use our high dimensional IF approach to determine if B cells within the graft are interacting with relevant T cell populations and we will use laser capture microscopy to isolate individual B cells from which we will clone and express the antigen combining regions to generate antibodies and then test specificity for different types of graft antigens. We will also modify our in vitro TEM assays to allow endothelial cells to cross-link and signal through the B cell receptor for antigen by display of anti-human kappa antibody, again employing a technique we used to mimic T cell receptor signaling with anti-human CD3. We anticipate that these studies will lead to identification of targets, possibly inhibitable by currently approved agents, that can limit B cell recruitment into allografts, an intervention that may reduce late graft loss without the risk to immunosuppressed patients mediated by total B cell ablation.

7. 项目概要/摘要由于慢性排斥导致的晚期同种异体移植肾丢失率在过去三年中没有显着变化几十年来，尽管在控制急性排斥反应方面取得了显着进展。虽然免疫发病机制是目前尚不完全清楚，相当一部分同种异体移植物含有浸润的宿主 B 细胞，并且存在 B 细胞的减少与不良预后相关，这可能与供体特异性抗体的存在无关。虽然 T 细胞向同种异体移植物的募集已被充分了解，并且可能由趋化因子或抗原启动血管腔内的呈现，对于 B 细胞的募集知之甚少。我们建议学习这些过程的目的是确定可能抑制 B 细胞浸润的途径和分子靶点移植物。为了研究趋化因子是否以及如何诱导这一过程，我们将采用高维人同种异体移植组织的循环组织免疫荧光显微镜和跨内皮细胞的体外测定人外周血 B 细胞的迁移 (TEM)。组织分析将提供有关亚型的信息移植物浸润中发现的 B 细胞、它们表达的趋化因子和粘附受体以及它们与激活的内皮细胞是可能的进入点。体外试验将使用人体微血管微流体室中的内皮细胞单层，用于表征和操纵能够使用我们在 T 细胞研究中开发的方法进行 TEM。研究这个过程是否以及如何进行由抗原诱导，我们将使用我们的高维 IF 方法来确定移植物内的 B 细胞是否与相关 T 细胞群相互作用，我们将使用激光捕获显微镜分离单个 B 细胞我们将克隆并表达抗原结合区以产生抗体，然后进行测试对不同类型移植物抗原的特异性。我们还将修改我们的体外 TEM 测定，以允许内皮细胞细胞通过 B 细胞受体通过展示抗人 kappa 抗体进行交联并发出抗原信号，再次采用我们用抗人 CD3 来模拟 T 细胞受体信号传导的技术。我们预计这些研究将导致目标的确定，可能被目前批准的药物抑制，可以限制 B 细胞募集到同种异体移植物中，这种干预措施可以减少晚期移植物丢失，而没有风险由总 B 细胞消融介导的免疫抑制患者。