Study of Human B-cell Tolerance by Functional Genomics

功能基因组学研究人类 B 细胞耐受性

• 批准号: 6883230
• 负责人: Ignacio E. Sanz
• 金额: $ 23.63万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-15 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6883230
• 关键词: B cell receptor; B lymphocyte; CD1 molecule; T lymphocyte; anergy; autoantigens; autoimmunity; biological signal transduction; cell cell interaction; cellular immunity; clinical research; flow cytometry; functional /structural genomics; gene expression; gene expression profiling; glycolipids; helper T lymphocyte; human subject; immune tolerance /unresponsiveness; immunocytochemistry; microarray technology; pathologic process; phenotype; spleen; systemic lupus erythematosus; toll like receptor

DESCRIPTION (provided by applicant): This R21 application is submitted in response to PAS-02-160 (Application of Exploratory/Developmental Technologies to NIAID-Funded Research) on the basis of our RO1 grant # AI 49660-01A1. A central problem for the study of human B-cell tolerance is the lack of suitable experimental systems. We have approached this critical problem by establishing that a subset of autoreactive human B-cells (VH4.34 B-cells) is strictly censored in healthy subjects from participating in productive germinal center (GC) reactions. In addition, we have shown that the regulatory mechanisms acting upon VH4.34 B-cells in healthy subjects are faulty in SLE patients, where VH4.34 B-cells are frequently found in the germinal centers as well as in the IgG memory and PC compartments and contribute substantially to the pathogenic autoantibody repertoire. Another major problem in this field has to do with the complex nature of the regulatory processes responsible for the maintenance of B-cell tolerance. Indeed, the fate of autoreactive B-cells depends on the balance of multiple signaling (both positive and negative), survival and migration programs with the ability to mutually regulate one another through modulatory and feedback circuits. While targeted biochemical, cellular and genetic approaches are useful to reveal the role of specific factors and pathways, we submit that a functional genomics approach can provide unique insights into the role and interaction of known players and may also identify previously unsuspected elements. We have recently been funded through the RO1 mechanism to study different aspects of human B-cell tolerance using VH4.34 B-cells as an experimental system. In this R21 application, we propose to extend these studies by using DNA microarray analysis to compare the gene expression profiles of naive autoreactive VH4.34 B-cells in normal individuals with those of control naive B-cells. In addition, we shall also study the gene expression profiles of SLE VH4.34 B-cells before and during their transition to germinal center B-cells. Our experimental approach has the potential to identify genes generally associated with SLE as well as genes specifically involved in the maintenance of B-cell tolerance. In addition, although the questions asked are hypothesis-driven, the approach proposed is highly exploratory and, as such, is responsive to this RFA.

描述（由申请人提供）：根据我们的RO1赠款＃AI 49660-01A1，该R21申请是针对PAS-02-160（将探索性/发展技术应用于NIAID资助的研究的应用）提交的。 研究人B细胞耐受性的一个核心问题是缺乏合适的实验系统。我们已经通过确定自动反应性人B细胞（VH4.34 B细胞）的子集严格审查健康受试者，从参与生产生发中心（GC）反应中严格审查。此外，我们已经表明，在健康受试者中作用于VH4.34 b细胞的调节机制在SLE患者中是错误的，在该患者中，VH4.34 B细胞在生发中心以及IgG记忆和PC室中经常发现，并为致病性自动抗体捕获量库做出了实质性贡献。 该领域的另一个主要问题与负责维持B细胞公差的监管过程的复杂性有关。实际上，自动反应性B细胞的命运取决于多个信号传导（正面和负面），生存和迁移计划的平衡，具有通过调节和反馈电路相互调节的能力。虽然有针对性的生化，细胞和遗传方法对于揭示特定因素和途径的作用很有用，但我们认为功能性基因组学方法可以为已知玩家的作用和相互作用提供独特的见解，并且也可能识别先前未知的元素。 最近，我们通过RO1机制资助了使用VH4.34 B细胞作为实验系统研究人B细胞耐受性的不同方面。在此R21应用中，我们建议通过使用DNA微阵列分析来扩展这些研究，以比较具有控制的天真B细胞的正常个体中天真自动反应性VH4.34 B细胞的基因表达谱。此外，我们还将研究SLE VH4.34 B细胞在过渡到生发中心B细胞之前和期间的基因表达谱。我们的实验方法具有鉴定通常与SLE相关的基因以及特别涉及B细胞耐受性的基因。此外，尽管提出的问题是由假设驱动的，但提出的方法是高度探索性的，因此，对此RFA有反应。