The Mouse Blood Center Core

项目摘要 背景：在输血和妊娠环境中，红细胞异体免疫是一个临床上重要的问题。 一模一样。考虑到人体研究中涉及的变量的数量，这个PPG中的项目已经采取了 使用简化主义小鼠模型更好地全面理解因素的方法 影响红细胞同种异体免疫。这一提议的老鼠血液中心核心(核心B)将支持4 PPG 通过提供用于输血的降低白血球、质量控制的转基因红细胞的项目。此外，这一核心将 作为同种异体抗体检测的中央免疫血液学参考实验室。除了优化 资源利用率和效率，该核心提供的产品和服务将允许更直接 场地和项目之间的结果比较。 创新：建议在这些研究中使用的小鼠供体品系，由 PPG研究人员允许隔离单变量变化对同种免疫的影响。首置设备 捐献者的红细胞上有一个三重融合蛋白，由鸡蛋溶菌酶、卵清蛋白和 人类Duffyb抗原。KEL捐献者在RBC中表达了整个人类KEL糖蛋白 具体方式，本提案中使用了K1和K2捐助者。野生型产生的抗体 输注转基因HOD、K1或K2红细胞的受体小鼠具有临床意义，导致 胎儿和新生儿的红细胞过早清除、溶血性输血反应和溶血性疾病 (HDFN)。 具体目标1：培育和维持表达模型或人类血型的转基因小鼠 红血球上的抗原。 具体目标2：收集、加工、执行质量控制措施并分发捐赠者红细胞 从这些转基因动物转移到所有4个PPG位点。 具体目标3：作为抗体检测的免疫血液学参考实验室。

Project Summary Background: RBC alloimmunization is a clinically significant problem in transfusion and pregnancy settings alike. Given the number of variables involved in human studies, the projects in this PPG have taken the approach of using reductionist murine models to generate a better overall understanding of factors influencing RBC alloimmunization. This proposed Mouse Blood Center Core (Core B) will support the 4 PPG projects by providing leukoreduced, quality controlled, transgenic RBCs for transfusion. Further, this Core will serve as a central immunohematology reference Laboratory for alloantibody testing. In addition to optimizing resource utilization and efficiency, the products and services provided by this Core will allow for more direct comparison of results between sites and projects. Innovation: The mouse donor strains proposed for use in these studies, generated and characterized by PPG investigators, allow for isolation of the effects of single variable changes on alloimmunization. The HOD donors have a triple fusion protein on their RBCs, composed of hen egg lysozyme, ovalbumin, and the human Duffyb antigen. The KEL donors have the entire human KEL glycoprotein expressed in an RBC specific manner, with K1 as well as K2 donors being used in this proposal. Antibodies generated by wild type recipient mice transfused with transgenic HOD, K1, or K2 RBCs are clinically significant, resulting in premature RBC clearance, hemolytic transfusion reactions, and hemolytic disease of the fetus and newborn (HDFN). Specific Aim 1: To breed and maintain transgenic mice expressing model or human blood group antigens on their RBCs. Specific Aim 2: To collect, process, perform quality control measures, and distribute donor RBCs from these transgenic animals to all 4 PPG sites. Specific Aim 3: To serve as an immunohematology reference laboratory for antibody detection.