Li X K, Okuyama T, Tamura A, Enosawa S, Kaneda Y, Takahara S, Funashima N, Yamada M, Amemiya H, Suzuki S
Department of Experimental Surgery & Bioengineering, National Children's Medical Research Center, Tokyo, Japan.
Transplantation. 1998 Dec 15;66(11):1416-23. doi: 10.1097/00007890-199812150-00003.
Transplantation of Fas ligand (FasL) gene-transfected tissues can have opposite effects. For example, cotransplantation of pancreas islets with myoblasts transfected with FasL-expressing plasmid vector (pFasL) prevented graft rejection, whereas the expression of FasL directly within islets using adenovirus vector led to graft destruction. It was also reported that FasL expression on pancreas islets led to neutrophilic infiltration and rapid destruction of the islets. From these results, overexpression of FasL in transfected tissues may lead directly to self destruction through an autocrine Fas-FasL pathway or graft destruction through neutrophil recruitment. To date there have been no reports of successful transplantation of FasL gene-transfected solid organs.
Rat pFasL was transfected at a dose of 90, 180, 270, or 360 microg into rat liver with an inactivated hemagglutinating virus of Japan conjugated to liposome vesicles (HVJ-liposome), and the gene-transfected livers were transplanted to allogeneic rats.
In 18 rats transfected with 180 microg of pFasL, 14 (78%) did not develop fulminant hepatitis. FasL-mRNA was detected in these livers at 3, 5, 7, and 14 days after transfection. The expression of FasL protein was also observed in the transfected liver, and the transfection rate by this method was 11.1+/-1.9%. The livers were then transplanted to allogeneic recipients, resulting in significant (P<0.01) prolonged recipient survival times. Histological observation showed that the pFasL-transfected liver allografts caused apoptotic cell death in infiltrating activated T cells. In contrast, transfection of pFasL higher than 180 microg resulted in lethal hepatitis in all rats, and its low dose (90 microg) did not induce the hepatitis or prolong recipient survival.
Our results indicate that rat liver allografts can be protected to host immune responses by an adequate level (approximately 10%) of FasL expression in the livers using HVJ-liposome incorporating pFasL.
转染Fas配体（FasL）基因的组织移植可能产生相反的效果。例如，将胰岛与转染了表达FasL质粒载体（pFasL）的成肌细胞共同移植可防止移植物排斥，而使用腺病毒载体在胰岛内直接表达FasL则导致移植物破坏。也有报道称胰岛上的FasL表达会导致中性粒细胞浸润和胰岛的快速破坏。从这些结果来看，转染组织中FasL的过表达可能通过自分泌Fas - FasL途径直接导致自我破坏，或通过募集中性粒细胞导致移植物破坏。迄今为止，尚无关于成功移植转染FasL基因的实体器官的报道。
将90、180、270或360微克剂量的大鼠pFasL用与脂质体囊泡偶联的日本灭活血凝病毒（HVJ - 脂质体）转染到大鼠肝脏中，然后将基因转染的肝脏移植到同种异体大鼠体内。
在18只转染180微克pFasL的大鼠中，14只（78%）未发生暴发性肝炎。在转染后3、5、7和14天在这些肝脏中检测到FasL - mRNA。在转染的肝脏中也观察到FasL蛋白的表达，该方法的转染率为11.1±1.9%。然后将肝脏移植到同种异体受体中，导致受体存活时间显著延长（P<0.01）。组织学观察表明，pFasL转染的肝脏同种异体移植物导致浸润的活化T细胞发生凋亡性细胞死亡。相比之下，高于180微克的pFasL转染导致所有大鼠发生致死性肝炎，其低剂量（90微克）未诱发肝炎或延长受体存活时间。
我们的结果表明，使用掺入pFasL的HVJ - 脂质体，通过肝脏中适当水平（约10%）的FasL表达，大鼠肝脏同种异体移植物可免受宿主免疫反应的影响。
