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在被称为生发中心的继发性淋巴组织的专业区域中进行T和B细胞协作。了解这些细胞相互作用的调节对于疫苗发育和了解生发中心淋巴细胞在肿瘤发育和自身免疫性中的作用很重要。 T卵泡辅助细胞（TFH）是能够迁移到卵泡并向B细胞传递辅助信号的专门CD4 T细胞，因此在体液免疫反应的功能成熟中起着至关重要的作用。基因表达的全球分析表明，生发中心B细胞和TFH均具有独特的基因表达模式，可以将它们与其他天真和活化的淋巴细胞的其他亚群区分开。据报道，其中一个基因在生发中心B细胞和TFH编码托克斯（Tox）中特别表达，TOX是啮齿动物和人之间高度保守的蛋白质，在我们的实验室中首次确定为T细胞发育的调节剂。我们已经产生了托克斯转基因小鼠以及全球和有条件的缺陷小鼠，并表明托克斯是CD4 T细胞谱系开发所必需的。使用这些转基因动物的组合，我们在这里建议确定托克斯是否也在生发中心反应中起作用。我们的方法使我们能够区分托克斯缺陷对B和T细胞的影响。因此，有条件地育种毒素缺陷的小鼠是在B细胞谱系中表达CRE重组酶的小鼠，将使用在免疫后寻找生发中心B细胞形成和功能的缺陷。提出了两种互补方法，以类似地研究托克斯在TFH发育和功能中的作用。在CD4+ T细胞中，与胸膜表达的TOX转基因和体外CRE介导的毒Tox基因座的缺失的全球毒素缺陷小鼠的互补，随后对TFH发育和在体内功能进行了测试。为了补充这些研究，我们还建议生成小鼠的托克斯报告基因敲入菌株，这将使我们能够在免疫反应期间跟踪托克斯在活体和体内的表达。公共卫生相关性：保护性抗体介导的免疫反应需要在称为生发中心的继发性淋巴组织的专业区域中进行T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的发育和理解生发中心淋巴细胞在肿瘤发育和自身免疫性中的作用很重要。该研究计划旨在了解特定的核蛋白质（在啮齿动物和人类中高度保守的）在调节这些生发中心反应中的作用。