CD180(RP105) Regulation of TLR and BCR Responses in B Cells

CD180(RP105) 对 B 细胞中 TLR 和 BCR 反应的调节

• 批准号: 7772774
• 负责人: JEFFREY A LEDBETTER
• 金额: $ 22.76万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7772774
• 关键词: Affinity; Agonist; Antibodies; Antibody Formation; Antigens; Apoptosis; B Cell Proliferation; B-Lymphocytes; Binding; Cancer Model; Cell Line; Cessation of life; Chimeric Proteins; Communicable Diseases; Complex; Couples; Data; Engineering; Fc domain; Future; Goals; Grant; Human; IgG3; Immunoglobulin Variable Region; Immunomodulators; In Vitro; Inflammatory; Injection of therapeutic agent; Interleukin-10; Interleukin-6; Lead; Ligands; Link; Mammalian Cell; Mediating; Membrane; Molecular; Mus; Mutate; Production; Proteins; Receptor Signaling; Receptors, Antigen, B-Cell; Recombinants; Regulation; Role; Serum; Signal Transduction; Signal Transduction Pathway; T-Lymphocyte; TLR2 gene; TLR4 gene; Tail; Testing; Toxic effect; Transgenic Mice; Transgenic Organisms; Tyrosine Phosphorylation; Work; antigen binding; autoreactive B cell; base; cytokine; design and construction; dimer; immunoregulation; in vivo; novel; public health relevance; receptor; research study; response; tandem mass spectrometry; therapeutic target; tool

DESCRIPTION (provided by applicant): This proposal will investigate the potential for B cell immunoregulation by CD180 stimulation and study the mechanisms of CD180 interactions with BCR and TLR signals. CD180 (RP105) expression is required for TLR4 and TLR2-mediated B cell proliferation and T cell- independent antibody responses (TI-1 response). We propose a comprehensive study of the CD180 receptor, including studies of the mechanisms of CD180 interaction with BCR and TLR signals, combined with analysis of CD180 activation in mice to determine the effect on antigen-specific B cells expressing transgenic high affinity BCR (B1-8hi) specific for NP, and on the antibody response to NP-CGG. These experiments will determine whether CD180 activation may serve as a sensitization signal that could result in selective B cell deletion upon antigen binding. CD180 is thought to coordinate or integrate the TLR4 and TLR2 signals with BCR signals in B cells to drive an antigen specific antibody response of all isotypes. We have found that CD180 stimulation together with TLR agonists results in synergistic B cell proliferation, and augments B cell production of inflammatory cytokines IL-6, IL-10, and TNF1. In contrast, proliferation of CD180-stimulated B cells is inhibited by simultaneous BCR stimulation, and CD180-activated B cells are sensitized to anti-BCR apoptosis. The second major component of this grant will use molecular engineering of soluble forms of CD180/MD-1 that are expressed in mammalian cells to help identify and potentially block natural CD180 ligands. We will also express anti-human and anti-mouse CD180 scFv-Ig fusion proteins. The scFvs will be constructed with wt or mutated human and mouse Ig tails in order to study the regulation of CD180 signals by FcR binding. The results will help clarify the role of CD180 as a master regulator of TLR2 and TLR4 and will help determine the potential for CD180 as a target for antibody-based therapy. The work will also result in construction and expression of recombinant molecules that will be useful in future studies of CD180 therapy in infectious disease and cancer models. PUBLIC HEALTH RELEVANCE: CD180 is a critical receptor for regulation of TLR4 and TLR2 function in B cells and could be an important target for immunoregulation. This grant will express recombinant molecules for CD180 stimulation and will test the potential for CD180 as a therapeutic target for deletion of autoreactive B cells.

描述(由申请人提供)：这项建议将研究CD180刺激对B细胞免疫调节的可能性，并研究CD180与BCR和TLR信号相互作用的机制。CD180(RP105)的表达是TLR4和TLR2介导的B细胞增殖和T细胞非依赖性抗体应答(TI-1应答)所必需的。我们建议对CD180受体进行全面的研究，包括CD180与BCR和TLR信号相互作用的机制的研究，并结合CD180在小鼠体内的激活分析，以确定CD180对表达针对NP的转基因高亲和力BCR(B1-8HI)的抗原特异性B细胞的影响，以及对NP-CGG的抗体应答的影响。这些实验将确定CD180的激活是否可以作为致敏信号，在抗原结合时导致选择性B细胞缺失。CD180被认为在B细胞中协调或整合TLR4和TLR2信号与BCR信号，以驱动所有同型的抗原特异性抗体反应。我们发现CD180的刺激与TLR激动剂一起导致了B细胞的协同增殖，并增加了B细胞产生炎症细胞因子IL-6、IL-10和TNF1。相反，CD180刺激的B细胞的增殖被同时的bcr刺激抑制，CD180激活的B细胞对抗bcr的凋亡敏感。这笔赠款的第二个主要组成部分将使用在哺乳动物细胞中表达的可溶性形式的CD180/MD-1的分子工程来帮助识别和潜在地阻断天然的CD180配体。我们还将表达抗人和抗鼠CD180单链抗体-Ig融合蛋白。为了研究FCR结合对CD180信号的调节作用，我们将用重尾或突变的人和鼠Ig尾巴构建单链抗体。这一结果将有助于阐明CD180作为TLR2和TLR4的主要调节因子的作用，并将有助于确定CD180作为基于抗体的治疗靶点的潜力。这项工作还将导致重组分子的构建和表达，这将有助于未来CD180治疗传染病和癌症模型的研究。 公共卫生相关性：CD180是调节B细胞中TLR4和TLR2功能的关键受体，可能是免疫调节的重要靶点。这笔赠款将表达CD180刺激的重组分子，并将测试CD180作为删除自身反应性B细胞的治疗靶点的潜力。