Immune Tolerence to Transplanted Myoblasts

对移植的成肌细胞的免疫耐受

• 批准号: 7580970
• 负责人: DAVID M ROTHSTEIN
• 金额: --
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-26 至 2009-02-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7580970
• 关键词: Address; Allogenic; Allografting; Animal Model; Antigens; Autologous; Bone Marrow Transplantation; Cells; Cellular Infiltration; Characteristics; Chimerism; Data; Development; Dose; Duchenne muscular dystrophy; Dystrophin; Engraftment; Exhibits; Family; Future; Genes; Goals; Hematopoietic; Histocompatibility; Immune; Immune response; Immune system; Immunosuppression; Immunotherapeutic agent; Infection; Inflammatory; Integral Membrane Protein; Kidney; Lymphocyte; Malignant Neoplasms; Mediating; Modeling; Monkeys; Mus; Muscle; Muscle Fibers; Myoblasts; Organ; PTPRC gene; Peripheral; Physiology; Protein Isoforms; Protein Tyrosine Phosphatase; Proteins; Protocols documentation; Regulation; Research Personnel; Resistance; Rest; Role; Signal Transduction; Signaling Molecule; Skin Transplantation; Skin graft; Solid; Specificity; Stem cell transplant; T-Cell Activation; T-Lymphocyte; Tissue Transplantation; Toxic effect; Transplantation; Treatment Protocols; Up-Regulation; Western Blotting; Whole-Body Irradiation; base; conditioning; cytokine; design; functional improvement; gene therapy; graft vs host disease; human disease; immunogenic; immunogenicity; insight; irradiation; islet; islet allograft; kidney allograft; mdx mouse; muscle strength; novel; novel strategies; novel therapeutics; pre-clinical; primary outcome; programs; research study; retransplantation; skin allograft; stem

The goal of this project is to treat experimental Duchenne Muscular Dystrophy using allogeneic myoblast transplantation without sustained immunosuppression through allogeneic myoblast transplantation (MT). This will require the induction of immunological tolerance towards allogeneic myoblasts, as well as to neoantigens resulting from the fusion of donor myoblasts with host muscle fibers (including dystrophin expressed by donor cells bearing the wild-type gene). MT in mice, has proved resistant to potent tolerogenic strategies that are successful in other transplantmodels. For this reason, we will utilize newly emerging strategies that incorporate mAbs against the higher Mr isoforms of the CD45 protein tyrosine phosphatase (anti-CD45RB). A short course of anti-CD45RB can induce tolerance to murine renal and islet allografts and can promote long-term engraftment of renal allografts in monkeys. Moreover, in combination with anti-CD40L, anti-CD45RB can induce prolonged engraftment of murine skin grafts in highly immunogenic strain combinations. Here, we will utilize anti-CD45RB to help induce central tolerance to MT in dystrophic mdx mice. Our preliminary data reveals that the combination of anti-CD45RB with anti-CD154 and bone marrow transplantation (BMT) induces stable high level mixed chimerism (i.e. co-existence of both donor and recipient hematopoietic cells) and allows robust tolerance toward transplanted myoblasts. In this regard, we obtain >100 day survival of allogeneic myoblasts in dystrophic mice using this approach, with high levels of dystrophin expression. In Aim 1, we will extend and refine our protocol in attempts to minimize toxicity of the conditioning regimen required to achieve mixed chimerism. Specific tolerance will be demonstrated by retransplantation of both same strain and different strain ("third-party") myoblasts. Improvement in muscle physiology will be assessed. In Aim 2, we will determine the mechanisms by which anti-CD45RB and BMT contribute towards tolerance in this model, based on our current understanding. Studies will include:determination of the specific immunological reactivity towards both allogeneic and neoantigens including dystrophin; demonstrating a shift in CD45 isoform expression; determining the role of CTLA-4 upregulation; induction of regulatory T cells and altered inflammatory cytokines; altered humoral immune responses; and the requirement for chimerism and thymic deletion. The results of these studies will contribute to development of an approach towards treating this uniformly fatal human disease. Moreover, an understanding of the immune response towards dystrophin, expressed at higher levels and in its native form, will be attained.

该项目的目的是使用同种异体成肌细胞治疗实验性Duchenne肌肉营养不良 通过同种异体成肌细胞移植（MT）的移植而无需持续免疫抑制。这会 需要对同种异体成肌细胞以及对新抗原的诱导免疫耐受性 从供体成肌细胞与宿主肌肉纤维的融合（包括由供体细胞表达的肌营养物 带有野生型基因）。事实证明，小鼠中的MT对成功的有效耐受性策略具有抵抗力 其他移植模型。因此，我们将利用新兴的策略，将mab纳入反对 CD45蛋白酪氨酸磷酸酶（抗CD45RB）的较高MR同工型。一小段抗CD45RB可以 诱导对鼠肾脏和胰岛同种异体移植的耐受性，并可以促进肾脏同种异体的长期植入 猴子。此外，结合抗CD40L，抗CD45RB可以诱导长时间的鼠类植入 高度免疫原性应变组合中的皮肤移植物。在这里，我们将利用抗CD45RB来帮助诱导中央 营养不良的MDX小鼠对MT的耐受性。我们的初步数据表明，抗CD45RB与 抗CD154和骨髓移植（BMT）诱导稳定的高水平混合嵌合体（即共存 供体和受体造血细胞），并允许对移植的肌细胞耐受性。在这个 考虑到，我们使用这种方法获得了营养不良小鼠中同种异体成肌细胞的生存> 100天 肌营养不良蛋白表达的水平。在AIM 1中，我们将扩展和完善我们的协议以最大程度地减少毒性 实现混合嵌合需要的条件方案。具体的公差将由 相同应变和不同菌株（“第三方”）成肌细胞的重新植入。肌肉的改善 生理学将进行评估。在AIM 2中，我们将确定抗CD45RB和BMT的机制 基于我们当前的理解，在此模型中促进宽容。研究将包括：确定 对包括肌营养不良蛋白在内的同种异体和新抗原的特异性免疫反应性； 证明CD45同工型表达的变化；确定CTLA-4上调的作用；诱导 调节性T细胞和炎性细胞因子改变；体液免疫反应改变；以及要求 嵌合和胸腺缺失。这些研究的结果将有助于发展一种方法 治疗这种统一致命的人类疾病。此外，了解对免疫反应的理解 肌营养不良蛋白以较高水平和本机形式表达。