Junker K, Koehl U, Zimmerman S, Stein S, Schwabe D, Klingebiel T, Grez M
Department of Pediatric Hematology and Oncology, University Hospital, Johann-Wolfgang von Goethe University, Frankfurt, Germany.
Gene Ther. 2003 Jul;10(14):1189-97. doi: 10.1038/sj.gt.3301977.
Donor lymphocyte infusions (DLI) following allogeneic stem cell transplantation are known to mediate graft-versus-leukemia effect (GVL). A major side effect of these immunotherapies is the development of graft-versus-host diseases (GVHD). One promising approach to prevent GVHD is to genetically modify donor T cells with a suicide mechanism that can be induced in the case of GVHD. Here we report on a retroviral vector containing the death effector domain (DED) of the human Fas-associated protein with death domain (FADD). The DED was fused to two copies of an FKBP506-binding protein and a truncated version of the human low-affinity receptor for nerve growth factor (LNGFR). Activation of the death signal pathway can be triggered upon the addition of chemical inducers of dimerization. This construct was functionally compared to an optimized HSV-TK vector in which a hypersensitive mutant of the herpes simplex virus thymidine kinase gene (TK39) was fused to a cytoplasmic truncated version of the cell surface antigen CD34. A direct comparison between both vectors in primary T lymphocytes showed that the number of T cells transduced with vectors containing the DED was significantly reduced within 24 h of drug administration whereas ganciclovir treatment of TK39-transduced T cells showed a delay in cell death of approximately 3-4 days. Our results indicate that constructs containing the DED may prove to be the most efficient mechanism to quickly eliminate alloreactive T cells.
同种异体干细胞移植后的供体淋巴细胞输注（DLI）已知可介导移植物抗白血病效应（GVL）。这些免疫疗法的一个主要副作用是移植物抗宿主病（GVHD）的发生。一种有前景的预防GVHD的方法是用一种自杀机制对供体T细胞进行基因改造，该机制可在发生GVHD时被诱导。在此，我们报告一种含有人类死亡结构域相关蛋白（FADD）死亡效应结构域（DED）的逆转录病毒载体。DED与两个FKBP506结合蛋白拷贝以及人类神经生长因子低亲和力受体（LNGFR）的截短版本融合。加入化学二聚诱导剂后可触发死亡信号通路的激活。将该构建体与一种优化的单纯疱疹病毒胸苷激酶（HSV-TK）载体进行功能比较，在HSV-TK载体中，单纯疱疹病毒胸苷激酶基因的一个超敏突变体（TK39）与细胞表面抗原CD34的胞质截短版本融合。在原代T淋巴细胞中对这两种载体进行直接比较显示，在给药后24小时内，用含有DED的载体转导的T细胞数量显著减少，而对TK39转导的T细胞进行更昔洛韦处理显示细胞死亡延迟约3 - 4天。我们的结果表明，含有DED的构建体可能被证明是快速消除同种异体反应性T细胞的最有效机制。
