GENE THERAPY IN INDUCTION OF TRANSPLANT TOLERANCE

尽管最近取得了成功，但临床进展的一个主要障碍 移植继续存在同种异体排斥反应和发病率， 非特异性免疫抑制导致的死亡率。因此， 实现持久的捐赠者特定的对外国的无反应 抗原将构成一项重要的临床进步。这是第一次 提案将审查基因转移技术的潜在用途，以 诱导抗原特异性移植耐受。更准确地说，这些 研究将确定是否可以致命地诱导耐受性 重组自体骨髓细胞的受照小鼠 哪些异种MHC基因已经被引入和表达。 含有人类人类白细胞抗原A2基因的逆转录病毒表达载体将 被构建并引入包装细胞系。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，获得最佳基因 调职。然后将受辐射的C57B1/6(B6)小鼠移植到 转化自体骨细胞表达人人类白细胞抗原A2基因。 然后对长期骨髓重组B6小鼠进行检查 1)体外免疫学对人类白细胞抗原A2的免疫反应性 用淋巴细胞刺激细胞系或转基因的检测 在B6背景上表达人类白细胞抗原A2基因的小鼠和2)通过 人类白细胞抗原A2转基因或第三方皮肤移植 对照供鼠移植到重组B6受体上。成功者 这些实验的完成应该会为我们深入了解异种生物 并将为进一步设计工作提供动力 研究跨越更复杂的组织相容障碍的耐受性 用于临床应用。

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.