Role of Nuclear Factor TOX in Germinal Center Reactions

核因子 TOX 在生发中心反应中的作用

基本信息

批准号：
7687579
负责人：
JONATHAN G KAYE
金额：
$ 20.31万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-16 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7687579
关键词：
Activated Lymphocyte Affinity Antibodies Antibody Formation Area Autoimmunity B-Lymphocytes Breeding CD4 Positive T Lymphocytes Cell Lineage Cells Collaborations Complement Defect Development Gene Expression Generations Genes Genetically Modified Animals Helper-Inducer T-Lymphocyte Human Immune response Immunization Immunoglobulin Class Switching Immunoglobulin Switch Recombination Immunoglobulins In Vitro Investigation Knock-in Mouse Laboratories Lymphocyte Lymphoid Tissue Mediating Mouse Strains Mus Nuclear Nuclear Protein Nuclear Proteins Pattern Play Proteins Reaction Regulation Reporter Reporting Research Rodent Role Signal Transduction Structure of germinal center of lymph node T-Cell Development T-Lymphocyte Testing Thymus Gland Transgenes Transgenic Mice base cell growth regulation in vivo programs public health relevance recombinase tool tumor vaccine development

项目摘要

DESCRIPTION (provided by applicant): The generation of a high affinity protective antibody response requires T and B cell collaboration in specialized areas of secondary lymphoid tissue known as germinal centers. Understanding the regulation of these cellular interactions is important for vaccine development and for understanding the role of germinal center lymphocytes in tumor development and autoimmunity. T follicular helper cells (TFH) are specialized CD4 T cells that are able to migrate into follicles and deliver helper signals to B cells, and thus play a critical role in functional maturation of a humoral immune response. Global analysis of gene expression has shown that both germinal center B cells and TFH have unique patterns of gene expression that distinguish them from other subsets of naive and activated lymphocytes. One of the genes reported to be expressed specifically in both germinal center B cells and TFH encodes TOX, a protein highly conserved between rodents and humans and first identified in our laboratory as a regulator of T cell development. We have produced TOX transgenic mice and globally and conditionally TOX-deficient mice and have shown that TOX is required for development of the CD4 T cell lineage. Using combinations of these genetically modified animals, we propose here to determine whether TOX also plays a role in germinal center reactions. Our approaches allow us to distinguish the effects of TOX-deficiency on B and T cells. Thus, breeding conditionally TOX-deficient mice to mice that express Cre recombinase in the B cell lineage will be used to look for defects in germinal center B cell formation and function upon immunization. Two complementary approaches are proposed to study similarly the role of TOX in TFH development and function; complementation of globally TOX-deficient mice with a thymically expressed TOX transgene and in vitro Cre-mediated deletion of the Tox locus in CD4+ T cells that are subsequently tested for TFH development and function in vivo. To complement these studies we also propose to generate a TOX reporter knock-in strain of mice that will allow us to track expression of TOX in viable cells and in vivo during an immune response. PUBLIC HEALTH RELEVANCE: A protective antibody-mediated immune response requires T and B lymphocyte interactions in specialized areas of secondary lymphoid tissue known as germinal centers. Understanding the regulation of these interactions is important for vaccine development and for understanding the role of germinal center lymphocytes in tumor development and autoimmunity. This research program is directed at understanding the role of a specific nuclear protein, highly conserved in rodents and humans, in regulating these germinal center reactions.

描述（由申请人提供）：高亲和力保护性抗体反应的产生需要 T 细胞和 B 细胞在称为生发中心的次级淋巴组织的特殊区域中协作。了解这些细胞相互作用的调节对于疫苗开发以及了解生发中心淋巴细胞在肿瘤发展和自身免疫中的作用非常重要。滤泡辅助 T 细胞 (TFH) 是专门的 CD4 T 细胞，能够迁移到滤泡中并向 B 细胞传递辅助信号，从而在体液免疫反应的功能成熟中发挥关键作用。基因表达的整体分析表明，生发中心 B 细胞和 TFH 都具有独特的基因表达模式，将它们与其他幼稚和活化淋巴细胞亚群区分开来。据报道，在生发中心 B 细胞和 TFH 中特异性表达的基因之一编码 TOX，这是一种在啮齿类动物和人类之间高度保守的蛋白质，首次在我们的实验室中被鉴定为 T 细胞发育的调节因子。我们已经培育出 TOX 转基因小鼠和全局性、条件性 TOX 缺陷小鼠，并证明 TOX 是 CD4 T 细胞谱系发育所必需的。使用这些转基因动物的组合，我们在此建议确定 TOX 是否也在生发中心反应中发挥作用。我们的方法使我们能够区分 TOX 缺陷对 B 细胞和 T 细胞的影响。因此，将条件性 TOX 缺陷小鼠与在 B 细胞谱系中表达 Cre 重组酶的小鼠进行繁殖，将用于寻找免疫后生发中心 B 细胞形成和功能的缺陷。提出了两种互补的方法来研究 TOX 在 TFH 发育和功能中的类似作用；用胸腺表达的 TOX 转基因和体外 Cre 介导的 CD4+ T 细胞中 Tox 位点的缺失来补充整体 TOX 缺陷的小鼠，随后在体内测试 TFH 的发育和功能。为了补充这些研究，我们还建议生成 TOX 报告基因敲入小鼠品系，这将使我们能够在免疫反应期间追踪活细胞和体内 TOX 的表达。公共卫生相关性：保护性抗体介导的免疫反应需要二级淋巴组织（称为生发中心）的特殊区域中 T 淋巴细胞和 B 淋巴细胞相互作用。了解这些相互作用的调节对于疫苗开发以及了解生发中心淋巴细胞在肿瘤发展和自身免疫中的作用非常重要。该研究计划旨在了解在啮齿动物和人类中高度保守的特定核蛋白在调节这些生发中心反应中的作用。