GENE THERAPY IN INDUCTION OF TRANSPLANT TOLERANCE

诱导移植耐受的基因治疗

• 批准号: 2066620
• 负责人: Ira J. Fox
• 金额: $ 8.93万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-01-01 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2066620
• 关键词: MHC class II antigen; Retroviridae; autologous transplantation; bone marrow transplantation; gene therapy; genetically modified animals; immune tolerance /unresponsiveness; immunogenetics; laboratory mouse; microspectrophotometry; polymerase chain reaction; radiation immunosuppression; skin transplantation; southern blotting; transfection; transplantation immunology

Despite recent successes, a major impediment to progress in clinical transplantation continues to be allograft rejection and morbidity and mortality resulting from non-specific immunosuppression. Therefore, achievement of long lasting donor-specific unresponsiveness to foreign antigens would constitute an important clinical advance. This FIRST proposal will examine the potential use of gene transfer technology to induce antigen-specific transplantation tolerance. More precisely, these studies will determine whether tolerance can be induced in lethally irradiated mice reconstituted with autologous bone marrow cells into which xenogeneic MHC genes have been introduced and expressed. Retroviral expression vectors, which contain the human HLA-A2 gene will be constructed and introduced into packaging cell lines. Colonies which produce high titer, replication-defective recombinant retroviruses will then be co-cultured with mouse bone marrow cells to attain optimal gene transfer. Irradiated C57B1/6 (B6) mice will then be transplanted with autologous bone cells transformed to express the human HLA-A2 gene. Long-term bone marrow reconstituted B6 mice will then be examined for immunologic reactivity to the HLA-A2 antigen by 1) in vitro immunologic assays using lymphocytes for stimulation from cell lines or transgenic mice expressing the HLA-A2 gene on the B6 background and 2) by transplantation of skin grafts from HLA-A2 transgenic or third-party control donor mice onto reconstituted B6 recipients. The successful completion of these experiments should provide insight into xenogeneic tolerance induction and will provide an impetus to further work designed to study tolerance across more complicated histocompatibility barriers for clinical application.

尽管最近取得了成功，但临床进展的主要障碍 移植仍然是同种异移植的排斥和发病率， 非特异性免疫抑制导致的死亡率。 所以， 实现持久的捐助者对外国人的反应 抗原将构成重要的临床前进。 第一个 提案将研究基因转移技术的潜在用途 诱导抗原特异性移植耐受性。 更确切地说，这些 研究将确定是否可以在致命中诱导公差 用自体骨髓细胞重构的辐照小鼠进入 已经引入并表达了哪些异构MHC基因。 包含人HLA-A2基因的逆转录病毒表达载体将 被构造并引入包装细胞系。 殖民地 产生高滴度，复制缺陷的重组逆转录病毒将 然后与小鼠骨髓细胞共培养以获得最佳基因 转移。 然后将辐照的C57B1/6（B6）小鼠移植 自体细胞转化为表达人HLA-A2基因。 然后将检查长期骨髓重构B6小鼠的小鼠 1）体外免疫学对HLA-A2抗原的免疫反应性 使用淋巴细胞从细胞系或转基因刺激的测定 在B6背景上表达HLA-A2基因的小鼠和2） 从HLA-A2转基因或第三方移植皮肤移植 将供体小鼠控制到重建的B6接受者。 成功 这些实验的完成应提供对异质性的见解 耐受性诱导，并将为设计的进一步工作提供动力 研究更复杂的组织相容性障碍的耐受性 用于临床应用。