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: 8434108
• 负责人: Max Dale Cooper
• 金额: $ 28.42万
• 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/8434108
• 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.

描述（申请人提供）：这项研究的基础是这样的发现，即相互作用的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，尽管无颌脊椎动物（七鳃鳗和八目鳗）通过重组转化不完全生殖系VLR基因产生用于抗原识别的可变淋巴细胞受体（VLR）（VLRA、VLRB和VLRC）转化为完全组装的VLR基因。最近的研究表明，VLRA的组装和表达与胞苷脱氨酶1（CDA 1）的表达仅在淋巴上皮胸腺样鳃区，而VLRB和CDA 2的表达主要发生在造血组织。 提出了一个全面的分析，以阐明的发展，分布和功能的七鳃鳗VLRA和VLRC淋巴细胞谱系的比较，与B细胞样VLRB淋巴细胞，以测试的假设，VLRA和VLRC谱系代表agnathan T细胞类似物的gnathostome 1/2和3/4 T细胞的同种异体识别责任。 一个长期的目标是确定七鳃鳗组织相容性抗原，以测试VLRA和VLRC识别这些抗原以实现自我与非自我区分的假设。 第一个具体目的是确定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，以深入了解淋巴细胞谱系的功能和合作潜力。 第二个具体目的是纯化VLRA+、VLRB+和VLRC+淋巴细胞群体，并对其每种转录组进行比较分析，以获得关于细胞因子、趋化因子、细胞因子/趋化因子受体、溶细胞组分、VLR共受体候选物、共刺激分子、信号传导元件、和转录因子的三种类型的淋巴细胞在其静止和激活状态。 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应答能力。初步体内和体外研究表明，VLRA+和VLRC+淋巴细胞优先应答同种异体白色血细胞。 将鉴定刺激细胞类型，还将确定皮肤同种异体移植物排斥的细胞参与者，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应答与非应答淋巴细胞的基因表达谱和VLR序列。 将诱导VLRB同种抗体，并将来自同种抗原应答细胞的VLRA和VLRC受体工程化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。