Adachi K, Fujino M, Kitazawa Y, Funeshima-Fuji N, Takahara S, Kimura H, Li X-K
Laboratory of Transplantation Immunology, National Research Institute for Child Health and Development, Tokyo, Japan.
Transplant Proc. 2006 Oct;38(8):2710-3. doi: 10.1016/j.transproceed.2006.08.011.
Modulation of donor organs by transfection of a gene encoding immmunosuppresive molecules has been recognized as a less toxic approach to prevent allograft rejection. Fas-ligand (FasL) plays a critical role in activation-induced cell death of activated cytotoxic lymphocytes. This may provide a potential for induction of "immune privileged sites" to escape the host immune surveillance system. Cytokine response modifier A (CrmA), a gene product of cowpox virus, blocks caspase as well as perforin/granzyme-mediated apoptotic pathways. Therefore, it may suppress intragraft apoptosis. The aim of the present study was to investigate whether transfection of FasL or CrmA genes prolonged the survival of rat liver allografts. Using the high responder rat combination of DA (RT-1(a)) donor to LEW (RT-1(1)) recipient, we performed orthotopic liver transplantation with subsequent delivery of adenoviral vectors containing FasL, CrmA, or LacZ, at a dose of 1 x 10(9) pfu via a recipient tail vein using a Cre-mediated gene expression system. Recipient survival was assessed as well as immunohistochemical examination of the grafts for anti-CD2, TUNEL, and H&E staining. Statistical analysis was performed with the Mann-Whitney U test. The therapeutic groups showed significantly prolonged recipient survival compared with the LacZ-treated control group. Histologic analysis revealed reduced hepatocyte apoptosis in the CrmA-treated group and increased apoptosis of infiltrating mononuclear cells in the FasL-treated group. These data suggested that FasL and CrmA may be potent genes to prolong rat liver allograft survival.
通过转染编码免疫抑制分子的基因来调节供体器官，已被认为是一种毒性较小的预防同种异体移植排斥反应的方法。Fas配体（FasL）在活化的细胞毒性淋巴细胞的活化诱导细胞死亡中起关键作用。这可能为诱导“免疫特权部位”以逃避宿主免疫监视系统提供潜力。细胞因子反应调节因子A（CrmA）是牛痘病毒的基因产物，可阻断半胱天冬酶以及穿孔素/颗粒酶介导的凋亡途径。因此，它可能抑制移植内的细胞凋亡。本研究的目的是调查转染FasL或CrmA基因是否能延长大鼠肝同种异体移植的存活时间。使用DA（RT-1(a)）供体到LEW（RT-1(1)）受体的高反应性大鼠组合，我们进行了原位肝移植，随后通过Cre介导的基因表达系统，经受体尾静脉以1×10(9) pfu的剂量递送含有FasL、CrmA或LacZ的腺病毒载体。评估受体的存活情况，并对移植物进行抗CD2、TUNEL和苏木精-伊红染色的免疫组织化学检查。采用Mann-Whitney U检验进行统计分析。与LacZ处理的对照组相比，治疗组的受体存活时间显著延长。组织学分析显示，CrmA处理组的肝细胞凋亡减少，而FasL处理组的浸润单核细胞凋亡增加。这些数据表明，FasL和CrmA可能是延长大鼠肝同种异体移植存活时间的有效基因。
