Murine model of Rh incompatibility for alternative therapy and serologic reagents

替代疗法和血清试剂的 Rh 不相容小鼠模型

• 批准号: 7220450
• 负责人: DWANE E WYLIE
• 金额: $ 10.7万
• 依托单位: ERYTHROSITE, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7220450
• 关键词: Address; Alloimmunization; Alternative Therapies; Animal Model; Animals; Antibodies; Antigens; Area; Benchmarking; Binding; Biological Models; Blood; Blood Group Antigens; Blood group antibody D; Clinical; Complex; Computer Systems Development; Coombs' Test; Development; Erythrocytes; Exposure to; Facility Construction Funding Category; Female; Fetal Erythroblastosis; Fetus; Future; Genes; Genetic Polymorphism; Goals; Hepatitis B e Antigens; Homologous Gene; Human; Human Volunteers; Immune; Immune Tolerance; Immune response; Immune system; Immunization; Immunosuppression; Incidence; Individual; Infectious Agent; Investigation; Isoantibodies; Lead; Medicine; Methods; Modeling; Mouse Strains; Mus; Numbers; Partner in relationship; Patients; Phase; Prevention; Probability; Production; Proteins; Range; Reaction; Reagent; Rh Disease; Rh-Hr Blood-Group System; Rho(D) Immune Globulin; Risk; Serological; Shapes; Sickle Cell; Sickle Cell Anemia; Source; Specificity; Syndrome; System; Testing; Therapeutic immunosuppression; Transfusion; Transgenes; Transgenic Mice; Transgenic Organisms; Validation; Variant; clinically relevant; in vivo; mouse model; neonate; placental transfer; prevent; programs; success; therapy development

DESCRIPTION (provided by applicant): Antibodies that distinguish common Rh specificities cannot be induced in normal mice. Our hypothesis is that this is most likely due to the shaping of their immune repertoires by the murine Rh homologue, and that these specificities would be produced by mice whose repertoires are shaped in the presence of human Rh proteins. The goal of this project is to create strains of mice expressing human RH transgenes and use them to develop a mouse model of Rh incompatibility, which would have commercial potential in at least three areas: 1) Development of alternative therapies for treatment of Rh hemolytic disease of the newborn (HDN), a severe and potentially fatal syndrome, the prevention of which relies on a diminishing supply of human source material with ever-present risks from emerging or surreptitious infectious agents; 2) Production of murine monoclonal Rh serologic reagents with promise for specificity superior to those currently in use; and 3) Future development of reagents to prevent alloimmunization to blood group antigens. HDN has been treated successfully with Rh immune globulin since the 1960's. The success of the treatment, though, has reduced the Rh immune globulin supply by decreasing the number of individuals sensitized to the D antigen. For this reason, and because of the concern over the use of blood-derived products, alternative therapies are needed, but there is no reliable experimental system in which to develop and test them. The long-term goal of this project is to establish such a system for development of methods to not only prevent not only HDN, but to target prevention of alloimmunization to blood group antigens in general. Importantly, this project will provide a unique and accurate model system with the potential to study a broad range of clinically relevant issues in transfusion medicine, such as HDN, transfusion reactions, and IVIg therapy. In addition, the model could be used to address many basic questions in transfusion medicine dealing with composition and assembly of the erythrocyte Rh complex, immune tolerance, and immunosuppression. The proposal consists of two specific aims: 1. Construction of mice transgenic for the human RHD and RHce genes. Constructs containing human RHD and RHce cDNAs will be used for creation of the transgenic mice. When mice carrying the transgenes have been identified, their erythrocytes will be characterized for expression of human Rh blood group antigens. 2. Induction of Rh-specific antibodies and placental antibody transfer in RH transgenic mice. Rh-specific antibodies will be induced in the transgenic mice as proof of our hypothesis and the validity of this system as a model of HDN. The final validation of the system will be demonstration of placental transfer of D-specific antibodies in sensitized females by testing erythrocytes from their fetuses or neonates for bound anti-D by the direct antiglobulin test (DAT) and elution of anti-D. The long-term goal of this program is to develop alternatives treatments for preventing hemolytic disease of the newborn (HDN). This need exists because of the diminished supply of Rh immune globulin and concern that blood-derived products are not without risk. This project will provide a unique and accurate model system with the potential to study a broad range of clinically relevant issues in transfusion medicine, such as HDN, transfusion reactions, and IVIg therapy.

描述（由申请人提供）：在正常小鼠中不能诱导区分共同RH特异性的抗体。我们的假设是，这很可能是由于鼠类RH同源物对其免疫曲目的塑造，并且这些特异性将由小鼠产生，这些小鼠的曲目在存在人类RH蛋白的存在下形成了。该项目的目的是创建表达人类RH转基因的小鼠菌株并使用它们来开发鼠标不兼容的鼠标模型，至少在至少三个领域中具有商业潜力：1）开发用于治疗新生儿RH溶血性疾病（HDN）（HDN）（HDN）的替代疗法（HDN），一种严重且潜在的致命综合症，该综合症与人类的预测相关，以至于人类的预防或者是人类的预防，以至于供应的预测，以至于供应的预测，以至于人们对人类的预防物质产生了预测。传染性药物； 2）生产鼠单克隆RH血清学试剂，其特异性比当前使用的有望优于特异性； 3）未来开发试剂，以防止对血液组抗原的同种异体释放。自1960年代以来，HDN已成功地用RH免疫球蛋白治疗。但是，治疗的成功通过减少对D抗原的个体数量来减少RH免疫球蛋白供应。因此，由于人们对使用血液衍生产品的使用感到担忧，因此需要替代疗法，但是没有可靠的实验系统可以开发和测试它们。该项目的长期目标是建立这样一种方法，以开发方法，不仅不仅可以防止HDN，还可以靶向预防对血液组抗原的同种异体免疫化。重要的是，该项目将提供一个独特而准确的模型系统，具有研究在输血医学（例如HDN，输血反应和IVIG疗法）中广泛临床相关问题的潜力。此外，该模型可用于解决输血医学的许多基本问题，该问题处理了红细胞RH复合物，免疫耐受性和免疫抑制的组成和组装。该提案由两个具体目的组成：1。为人类RHD和RHCE基因的小鼠转基因的构造。包含人RHD和RHCE CDNA的构建体将用于创建转基因小鼠。当已经鉴定出携带转基因的小鼠时，它们的红细胞将被特征在于人类血液组抗原的表达。 2。诱导Rh转基因小鼠中RH特异性抗体和胎盘抗体转移。转基因小鼠将诱导RH特异性抗体，以证明我们的假设和该系统作为HDN模型的有效性。该系统的最终验证将是通过测试来自胎儿或新生儿的红细胞来证明D特异性抗体在敏化女性中的胎盘转移，或者通过直接的抗肿瘤蛋白测试（DAT）和抗D的抑制剂进行了抗D。该计划的长期目标是开发预防新生儿溶血疾病（HDN）的替代治疗方法。由于RH免疫球蛋白的供应量减少，并且担心血液来源的产品并非没有风险，因此存在这种需求。该项目将提供一个独特而准确的模型系统，并有可能研究输血医学中的广泛临床相关问题，例如HDN，输血反应和IVIG治疗。