TOLERANCE TO ALLOGRAFTS THROUGH GENE THERAPY

通过基因治疗对同种异体移植物的耐受性

• 批准号: 2687985
• 负责人: John J Iacomini
• 金额: $ 24.84万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2687985
• 关键词: MHC class I antigen; MHC class II antigen; Retroviridae; T cell receptor; bone marrow; cell mediated lymphocytolysis test; cell type; cytotoxic T lymphocyte; flow cytometry; gene expression; gene therapy; genetic manipulation; genetic transduction; genetically modified animals; helper T lymphocyte; homologous transplantation; laboratory mouse; major histocompatibility complex; mutant; polymerase chain reaction; skin transplantation; suppressor T lymphocyte; tissue /cell culture; transplant rejection; transplantation immunology

Induction of lymphohematopoietic chimerism through bone marrow transplantation (BMT) can lead to life-long transplantation tolerance without the need for chronic immunosuppression. However, BMT across major histocompatibility (MHC) barriers is severely limited by problems associated with the severity of the host preparative regimen required, the potential for lethal graft-versus-host disease (GVHD), and engraftment failure. These problems could potentially be avoided by using a gene therapy approach to introduce allogeneic MHC genes into autologous bone marrow rather than transferring immunocompetent cells into the host. Using retroviral gene therapy, the allogeneic MHC class I gene H-2Kb (Kb) has been introduced into syngeneic bone marrow. Reconstitution of lethally irradiated syngeneic mice with bone marrow transduced with the Kb gene has been found to produce the specific prolongation of Kb bearing skin allograft survival. The major goals of this proposal are to examine the mechanism(s) by which expression of the transduced allogeneic MHC gene in BM derived cells lead to prolonged survival of Kb disparate skin grafts, and to establish conditions permitting long-term expression of the transduced gene in BM derived cells for the purpose of inducing life-long allograft acceptance. The specific aims are to: 1) Establish conditions permitting long-term expression of the retrovirally transduced H-2Kb gene in bone marrow derived cells; 2) Determine the mechanism by which expression of the transduced MHC class I gene product affects reactivity of anti-Kb CD8 T cells. 3) Determine the mechanism by which expression of retrovirally transduced Kb in bone marrow derived cells affects the ability of CD4 T cells to respond Kb disparate targets; and, 4) Evaluate the ability of Kb expressing cells of various lineages to induce specific prolongation of Kb disparate skin graft survival. These studies should provide practical information important for the field of transplantation on the mechanism of tolerance induced by gene therapy, and should also provide theoretical insights which may advance our understanding of self-nonself discrimination with respect to MHC antigens.

通过骨髓诱导淋巴瘤的嵌合体 移植（BMT）可以导致终身移植耐受性 无需慢性免疫抑制。但是，BMT跨越 主要的组织相容性（MHC）障碍受到问题的严重限制 与所需的宿主准备方案的严重程度相关， 致命的移植物抗宿主病（GVHD）的潜力和 植入失败。 这些问题可能会避免 使用基因疗法方法将同种异体MHC基因引入 自体骨髓而不是转移免疫能力的细胞 进入主机。 使用逆转录病毒基因治疗，同种异体MHC类 I Gene H-2KB（KB）已被引入合成骨髓中。 用骨髓重建致命的辐照合元小鼠 已发现用Kb基因转导的特异性 KB轴承皮肤同种异体移植存活的延长。 主要目标 该提议是要检查表达的机制 在BM衍生细胞中转导的同种异体MHC基因导致延长 KB的生存不同的皮肤移植物，并确定条件 允许在衍生的BM中转导基因的长期表达 为了诱导终身同种异体移植的目的。 这 具体目的是：1）建立允许长期的条件 在骨髓中逆转录转导的H-2KB基因的表达 衍生细胞； 2）确定表达的机制 转导的MHC I类基因产物会影响抗KB CD8的反应性 T细胞。 3）确定逆转录病毒表达的机制 在骨髓衍生细胞中转导的Kb会影响CD4的能力 T细胞反应Kb不同的靶标； 4）评估能力 表达各种谱系的细胞的Kb诱导特定的细胞 KB延长皮肤移植物的生存。 这些研究应该 提供有关移植领域重要的实用信息 关于基因疗法诱导的耐受性的机制，也应 提供理论见解，可以提高我们对 关于MHC抗原的自称歧视。