Novel Innate Immune Receptors in Zebrafish

斑马鱼的新型先天免疫受体

• 批准号: 7316101
• 负责人: GARY WILLIAM LITMAN
• 金额: $ 33.72万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-15 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7316101
• 关键词: Address; Allogenic; Antigens; Biochemistry; Biological Assay; Biological Models; Biology; Catfish; Cell Line; Cell Lineage; Cell Surface Receptors; Cell surface; Cells; Channel Catfish; Characteristics; Complement Receptor; Development; Elements; Evolution; Exhibits; Family; Gene Cluster; Gene Expression; Gene Family; Genetic; Genomics; Green Fluorescent Proteins; Health; Immune; Immune system; Immunoglobulins; Immunologic Receptors; Immunological Models; Infection; Knowledge; Leukocytes; Ligand Binding; Ligands; Link; Malignant Neoplasms; Mammals; Mediating; Mediator of activation protein; Membrane; Modeling; Multigene Family; Natural Immunity; Natural Killer Cells; Numbers; Osteichthyes; Predisposition; Process; Receptor Gene; Recombinants; Regulation; Relative (related person); Reporter; Resistance to infection; Resolution; Role; Signal Transduction; Specificity; Staging; Structure; Structure-Activity Relationship; Surface; System; T-Cell Receptor; T-Lymphocyte; Technology; Transgenes; Transgenic Organisms; Zebrafish; base; developmental genetics; immune function; novel; novel strategies; receptor; response; structural biology; Address; Allogenic; Antigens; Biochemistry; Biological Assay; Biological Models; Biology; Catfish; Cell Line; Cell Lineage; Cell Surface Receptors; Cell surface; Cells; Channel Catfish; Characteristics; Complement Receptor; Development; Elements; Evolution; Exhibits; Family; Gene Cluster; Gene Expression; Gene Family; Genetic; Genomics; Green Fluorescent Proteins; Health; Immune; Immune system; Immunoglobulins; Immunologic Receptors; Immunological Models; Infection; Knowledge; Leukocytes; Ligand Binding; Ligands; Link; Malignant Neoplasms; Mammals; Mediating; Mediator of activation protein; Membrane; Modeling; Multigene Family; Natural Immunity; Natural Killer Cells; Numbers; Osteichthyes; Predisposition; Process; Receptor Gene; Recombinants; Regulation; Relative (related person); Reporter; Resistance to infection; Resolution; Role; Signal Transduction; Specificity; Staging; Structure; Structure-Activity Relationship; Surface; System; T-Cell Receptor; T-Lymphocyte; Technology; Transgenes; Transgenic Organisms; Zebrafish; base; developmental genetics; immune function; novel; novel strategies; receptor; response; structural biology

DESCRIPTION (provided by applicant): Leukocyte regulatory receptors (LRRs) effect a variety of immune functions, including self/nonself recognition and fine regulation of the response to infection. Membrane receptors belonging to several different LRR gene families are expressed by natural killer (NK) cells and detect a variety of ligands or absence thereof on the surfaces of different target cells. The full complement of receptors that mediate NK function is likely not understood. We have cloned the novel immune-type receptor (NITR) genes, which constitute a large multigene family encoding transmembrane immunoglobulin (Ig)-type molecules, from bony fish. NITRs exhibit structural characteristics of NK receptors and other LRRs, including activating/inhibitory-transmembrane signaling, and possess ectodomains of the Ig/T cell antigen receptor (TCR) variable (V) region type. NITR genes, which have been identified in several species of bony fish, including zebrafish (Danio rerio), do not undergo somatic reorganization and represent the only example of multigenic (cell surface) receptors outside of Ig and TCR genes in which highly diversified V regions likely are utilized as the core recognition elements of a receptor family. NITRs constitute a unique and potentially informative link between the innate and adaptive immune systems. Resolution of the complete zebrafish NITR gene cluster has identified multiple inhibitory forms and one activating form ofNITR. NITR V specificity for both endogenous and exogenous ligands will be examined using a number of different approaches based on: 1) soluble recombinant NITRs, 2) an NFAT-coupled GFP reporter assay for NITR ligand binding and 3) the expression of transgenic chimeric zebrafish/medaka (Oryzies latipes) Mhc I. A second model system, channel catfish (Ictalurus punctatus), in which NK-type cell lines that recognize allogenic targets have been derived, will be used for characterizing specific NITRtarget recognition. Information gained in the catfish system will be applied to identification of NITR-ligand interactions in zebrafish. The cell surface biochemistry of NITRs will be characterized and structural biology approaches will be used to compare NITR V regions to those of adaptive immune receptors. Knowledge of the full genomic structure of the zebrafish NITR cluster along with the relative ease of introducing transgenes and modulating expression levels of specific mRNAs in this species will be used to characterize both cell lineage- and developmental stage-specific expression of NITRs and to examine how NITR gene expression relates to innate immunity during early development. The unique advantages of zebrafish as a developmental model will permit us to examine vertebrate innate immunity prior to the initiation of adaptive immune function. Collectively, the multifaceted approach being utilized will define the essential structure/function relationships of a novel family of putative recognition molecules in a uniquely informative model system. Such information is essential to understanding the evolution and interrelatedness of NK and other LRR-mediated immune recognition processes, as well as the role of innate immunity during development. Understanding this relationship has broad health consequences in terms of mechanisms of resistance to infection and susceptibility to malignancy.

描述（由申请人提供）：白细胞调节受体（LRR）影响多种免疫功能，包括自我/非自然识别和对感染反应的良好调节。属于几个不同LRR基因家族的膜受体通过自然杀伤（NK）细胞表达，并在不同靶细胞的表面上检测多种配体或其不存在。介导NK功能的受体的完整补体很可能是无法理解的。我们已经克隆了新型的免疫型受体（NITR）基因，该基因构成了一个大型的多基因家族，该家族编码了来自骨鱼的跨膜免疫球蛋白（IG）型分子。 NITRS表现出NK受体和其他LRR的结构特征，包括激活/抑制 - 跨膜信号传导，并具有Ig/T细胞抗原受体（TCR）变量（V）区域类型的Ectodomain。在包括斑马鱼（Danio rerio）在内的几种骨鱼中已经鉴定出的NITR基因不经过体细胞重组，并且代表了Ig和TCR基因之外的多基因（细胞表面）受体的唯一例子，其中高度多样化的V区很可能被用作受体家族的核心识别元素。 NITRS构成了先天性和适应性免疫系统之间的独特且潜在的信息。完整斑马鱼基因簇的分辨率已鉴定出多种抑制形式和一种激活形式的nitr。 NITR V specificity for both endogenous and exogenous ligands will be examined using a number of different approaches based on: 1) soluble recombinant NITRs, 2) an NFAT-coupled GFP reporter assay for NITR ligand binding and 3) the expression of transgenic chimeric zebrafish/medaka (Oryzies latipes) Mhc I. A second model system, channel catfish （Ictalurus punctatus），其中识别同种异性靶标的NK型细胞系将用于表征特定的nitrtarget识别。在cat鱼系统中获得的信息将应用于斑马鱼中NITR-配体相互作用的识别。氮的细胞表面生物化学将被表征，结构生物学方法将用于比较氮V区与适应性免疫受体的区域。了解斑马鱼硝基簇的完整基因组结构，以及引入转基因的相对易于易于，并调节该物种中特定mRNA的表达水平，以表征细胞谱系和氮的发育阶段特异性表达，并检查NITR基因在早期发育过程中如何与天生的免疫相关。斑马鱼作为发展模型的独特优势将使我们能够在启动适应性免疫功能之前检查脊椎动物先天免疫。总体而言，使用的多方面方法将定义一个新颖的识别分子在独特的信息模型系统中的基本结构/功能关系。这种信息对于理解NK和其他LRR介导的免疫识别过程以及先天免疫在发育中的作用至关重要。了解这种关系在抵抗感染和对恶性肿瘤易感性的机制方面具有广泛的健康后果。