Small Animal Core

小动物核心

• 批准号: 8190136
• 负责人: YONG-GUANG YANG
• 金额: $ 36.82万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-15 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8190136
• 关键词: Animals; CD34 gene; CD47 gene; Cells; Chimera organism; Chimerism; Clinical; Engraftment; Family suidae; Fibrous capsule of kidney; Genes; Goals; Hematopoietic; Housing; Human; Immune system; Immunodeficient Mouse; Instruction; Laboratories; Ligands; Modeling; Mus; Natural Killer Cells; Organ Donor; Peptides; Protocols documentation; Reagent; Regulatory T-Lymphocyte; Research; Research Personnel; Research Project Grants; Role; Solutions; Stem cell transplant; Stem cells; T-Cell Development; Testing; Thymic Tissue; Thymus Gland; Tissues; Transgenes; Transgenic Mice; Transgenic Organisms; Transplantation; Xenograft procedure; cytokine; end-stage organ failure; fetal; mouse model; nonhuman primate; progenitor; programs; research study; stem; success

The role of Core C is to provide mouse models to all research projects. We will pursue two specific aims. Aim 1 includes: 1) coordinating, purchasing and housing immunodeficient (NOD/SCID and NOD/SCID/yc KO) mice, which will be used to prepare humanized mice (see Aim 2) for all projects (except Project 3, as explained in the Research Strategies section); and 2) maintaining and producing immunodeficient porcine cytokine transgenic mice. The porcine cytokine transgenic mice generated in this laboratory provide an excellent murine model for porcine hematopoietic stem/progenitor cell transplantation. These mice will be used to evaluate CD47 gene transduction in porcine hematopoietic progenitors (Project 2), and to prepare hu-mice for assessing human NK cell tolerance to porcine xenografts by mixed hematopoietic chimerism (Project 3). Aim 2 is to provide humanized mouse (hu-mouse) models, including: 1) 'standard' hu-mice with a functional human immune system; 2) hu-mice with porcine thymic grafts; 3) transgenic hu-mice that express human CD39 transgenes, and 4) porcine chimeric hu-mice with porcine hematopoietic cells expressing a HLA-E/human B2m/peptide trimer. Our approach to generating hu-mice with a functional immune system is to transplant fetal thymus tissue (under renal capsule) and CD34+ hematopoietic stem/progenitor cells (i.v.) in immunodeficient mice. The 'standard' hu-mice will be used to test the immunoregulatory reagents/protocols to be used in non-human primates (Projects 1 and 2). The HLAE/ human p2m/peptide trimer-expressing porcine chimeric hu-mice will be used to assess the potential of NK cell inhibitory ligand expression on porcine cells to induce human NK cell tolerance in Project 3. Hu-mice with porcine thymic grafts and human CD39-overexpresing hu-mice will be used in Project 4 to investigate the role of CD39 in human regulatory T cell development in xenogeneic porcine thymic grafts. The core will prepare and validate the mouse models, and then either perform the proposed experiments (for Projects 1 and 2) or provide 'ready-to-use' models to the investigators (for Projects 3 and 4). As detailed in each research components, these mouse models are essential to the success of this program project.

Core C的作用是为所有研究项目提供小鼠模型。我们将追求两个具体目标。 目标 1 包括：1) 协调、购买和饲养免疫缺陷（NOD/SCID 和 NOD/SCID/yc KO）小鼠，这些小鼠将用于为所有项目（项目 3 除外）制备人源化小鼠（参见目标 2），如研究策略部分）； 2)维持和产生免疫缺陷猪细胞因子转基因小鼠。本实验室培育的猪细胞因子转基因小鼠为猪造血干/祖细胞移植提供了优良的小鼠模型。这些小鼠将用于评估猪造血祖细胞中的CD47基因转导（项目2），并制备腐殖质小鼠，用于通过混合造血嵌合评估人类NK细胞对猪异种移植物的耐受性 （项目3）。目标2是提供人源化小鼠（hu-mouse）模型，包括：1）具有功能性人类免疫系统的“标准”hu-小鼠； 2) 带有猪胸腺移植物的腐殖质小鼠； 3) 表达人CD39转基因的转基因hu-小鼠，和4) 具有表达HLA-E/人B2m/肽三聚体的猪造血细胞的猪嵌合hu-小鼠。我们产生具有功能性免疫系统的 hu-小鼠的方法是将胎儿胸腺组织（肾囊下）和 CD34+ 造血干/祖细胞（静脉注射）移植到免疫缺陷小鼠中。 “标准”hu-小鼠将用于测试用于非人类灵长类动物的免疫调节试剂/方案（项目 1 和 2）。项目 3 中，表达 HLAE/人 p2m/肽三聚体的猪嵌合 hu-小鼠将用于评估猪细胞上 NK 细胞抑制性配体表达诱导人 NK 细胞耐受的潜力。 Hu-小鼠 项目 4 将使用猪胸腺移植物和人类 CD39 过度表达的 hu 小鼠来研究 CD39 在异种猪胸腺移植物中人类调节性 T 细胞发育中的作用。 核心将准备并验证小鼠模型，然后执行建议的实验（针对项目 1 和 2）或向研究人员提供“即用型”模型（针对项目 3 和 4）。正如每个研究部分所详述的，这些小鼠模型对于该计划项目的成功至关重要。