Definition of an Ancient T cell-like System in Lampreys

七鳃鳗中古代 T 细胞样系统的定义

• 批准号: 8219356
• 负责人: Max Dale Cooper
• 金额: $ 29.45万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8219356
• 关键词: Alloantigen; Allogenic; Antibodies; Antigen Receptors; Antigens; B-Lymphocytes; Binding; Cell physiology; Cells; Cellular Assay; Chimeric Proteins; Cytidine Deaminase; DNA Sequence Rearrangement; Data; Data Analyses; Detection; Development; Diagnostic Reagent; Discrimination; Elements; Engineering; Gene Expression; Gene Expression Profile; Genes; Genetic Polymorphism; Genomics; Gills; Goals; Hagfish; Hematopoietic; Histocompatibility Antigens; Immune; Immune system; Immunoglobulin Domain; Immunoglobulin Somatic Hypermutation; In Vitro; Infectious Agent; Isoantibodies; J segment gene; Jaw; Lampreys; Leucine-Rich Repeat; Leukocytes; Lymphocyte; Membrane; Modification; Molecular Profiling; Participant; Pattern; Phytohemagglutinins; Plasma Cells; Population; Receptor Gene; Receptors, Antigen, B-Cell; Recombinants; Signaling Molecule; System; T-Lymphocyte; Testing; Vertebrates; allograft rejection; analog; antigen binding; antigen processing; arm; base; cancer cell; cell type; chemokine; chemokine receptor; comparative; cytokine; design; hematopoietic tissue; histocompatibility gene; in vivo; insight; irradiation; receptor; response; skin allograft; transcription factor

DESCRIPTION (provided by applicant): The proposed studies build on the findings that interactive T- and B-like lymphocytes are basic features of the adaptive immune system of both jawless and jawed vertebrates, although jawless vertebrates (lampreys and hagfish) generate variable lymphocyte receptors (VLR) for antigen recognition by recombinatorial conversion of incomplete germline VLR genes (VLRA, VLRB and VLRC) into fully assembled VLR genes using diverse leucine-rich-repeat (LRR) donor sequences. Recent studies indicate that VLRA assembly and expression coincides with expression of cytidine deaminase 1 (CDA1) only within the lymphoepithelial thymoid gill region, whereas VLRB and CDA2 expression occurs primarily in hematopoietic tissues. A comprehensive analysis is proposed to elucidate the development, distribution and function of the lamprey VLRA and VLRC lymphocyte lineages in comparison with the B cell-like VLRB lymphocytes to test the hypothesis that the VLRA and VLRC lineages represent agnathan T cell analogues of gnathostome 1/2 and 3/4 T cells with allorecognition responsibility. A long term goal is to identify the lamprey histocompatibility antigens to test the hypothesis that VLRA and VLRC recognize these to achieve self versus non-self discrimination. The first specific aim is to define the distribution, antigen-binding and functional responses of the VLRA +, VLRB+ and VLRC+ lymphocytes and to modulate VLRA and VLRC cell function by treatment with VLR-specific antibodies to gain insight into the function and cooperative potential of the lymphocyte lineages. The second specific aim is to purify the VLRA+, VLRB+ and VLRC+ lymphocyte populations and perform a comparative analysis of each of their transcriptomes to obtain information about the differential expression of cytokines, chemokines, cytokine/chemokine receptors, cytolytic components, VLR co-receptor candidates, co-stimulatory molecules, signaling elements, and transcription factors for the three types of lymphocyte in their quiescent and activated states. The third specific aim is to characterize the allogeneic recognition and response capabilities of VLRA+, VLRB+ and VLRC+ lymphocytes. Pilot in vivo and in vitro studies indicate that the VLRA+ and VLRC+ lymphocytes preferentially respond to allogeneic white blood cells. The stimulatory cell types will be identified, the cellular participants will also be defined for skin allograft rejections, and the modulatory effects of VLRA- and VLRB-specific antibodies will be determined. Gene expression profiles and VLR sequences will be compared for alloantigen-responsive versus non-responsive lymphocytes. VLRB alloantibodies will be induced and VLRA and VLRC receptors from the alloantigen responding cells will be engineered to form secreted chimeric proteins for use in histocompatibility antigen detection. Candidate histocompatibility genes will be examined for genetic polymorphism, expression patterns and immunostimulatory potential. PUBLIC HEALTH RELEVANCE: These studies will yield insight into the basic evolutionary principles of self versus non-self recognition in vertebrates. They will also elucidate the functional interactions between the T and B cell arms of the lamprey immune system to facilitate development of lamprey VLRB antibodies for biomedical purposes by serving as unique diagnostic reagents for identifying infectious agents and cancer cells.

描述（由申请人提供）：拟议的研究建立在以下发现的基础上：尽管无颌脊椎动物（七鳃鳗和盲鳗）产生可变的淋巴细胞受体，但交互式 T 和 B 样淋巴细胞是无颌和有颌脊椎动物适应性免疫系统的基本特征(VLR) 通过使用不同的方法将不完整种系 VLR 基因（VLRA、VLRB 和 VLRC）重组转化为完全组装的 VLR 基因，从而进行抗原识别富含亮氨酸重复（LRR）供体序列。最近的研究表明，VLRA 的组装和表达仅在淋巴上皮胸腺鳃区域内与胞苷脱氨酶 1 (CDA1) 的表达一致，而 VLRB 和 CDA2 的表达主要发生在造血组织中。 提出了一项综合分析，以阐明七鳃鳗 VLRA 和 VLRC 淋巴细胞谱系的发育、分布和功能，并与 B 细胞样 VLRB 淋巴细胞进行比较，以检验 VLRA 和 VLRC 谱系代表颚口动物无颌 T 细胞类似物的假设 1/具有同种异体识别责任的 2 和 3/4 T 细胞。 长期目标是鉴定七鳃鳗组织相容性抗原，以检验 VLRA 和 VLRC 识别这些抗原以实现自我与非自我区分的假设。 第一个具体目标是定义 VLRA +、VLRB+ 和 VLRC+ 淋巴细胞的分布、抗原结合和功能反应，并通过用 VLR 特异性抗体处理来调节 VLRA 和 VLRC 细胞功能，以深入了解 VLRA +、VLRB+ 和 VLRC+ 淋巴细胞的功能和合作潜力。淋巴细胞谱系。 第二个具体目标是纯化VLRA+、VLRB+和VLRC+淋巴细胞群，并对它们各自的转录组进行比较分析，以获得细胞因子、趋化因子、细胞因子/趋化因子受体、溶细胞成分、VLR辅助受体候选物的差异表达信息、共刺激分子、信号元件和转录因子，适用于三种类型的淋巴细胞（处于静止和激活状态）。 第三个具体目标是表征 VLRA+、VLRB+ 和 VLRC+ 淋巴细胞的同种异体识别和反应能力。体内和体外试验研究表明，VLRA+ 和 VLRC+ 淋巴细胞优先响应同种异体白细胞。 将确定刺激细胞类型，还将定义皮肤同种异体移植排斥反应的细胞参与者，并确定 VLRA 和 VLRB 特异性抗体的调节作用。 将比较同种异体抗原反应性淋巴细胞与无反应性淋巴细胞的基因表达谱和 VLR 序列。 将诱导 VLRB 同种抗体，并且来自同种抗原应答细胞的 VLRA 和 VLRC 受体将被改造以形成分泌的嵌合蛋白，用于组织相容性抗原检测。 将检查候选组织相容性基因的遗传多态性、表达模式和免疫刺激潜力。 公共健康相关性：这些研究将深入了解脊椎动物自我识别与非自我识别的基本进化原理。 他们还将阐明七鳃鳗免疫系统的 T 细胞臂和 B 细胞臂之间的功能相互作用，通过充当识别感染因子和癌细胞的独特诊断试剂，促进用于生物医学目的的七鳃鳗 VLRB 抗体的开发。