Swenson K M, Ke B, Wang T, Markowitz J S, Maggard M A, Spear G S, Imagawa D K, Goss J A, Busuttil R W, Seu P
Dumont-UCLA Transplant Center, Department of Surgery, UCLA School of Medicine, Cedars-Sinai Medical Center, Los Angeles, California 90095, USA.
Transplantation. 1998 Jan 27;65(2):155-60. doi: 10.1097/00007890-199801270-00002.
Fas ligand (FasL) induces apoptosis of cells bearing its receptor Fas, and has been shown to be important in T-cell development and regulation and in immune privilege. We hypothesized that FasL expression by renal allografts might provide protection from rejection.
The murine FasL cDNA was cloned into a replication-defective adenovirus (AdV-FasL). Protein expression was confirmed by immunostaining of AdV-FasL-transduced HeLa cells. Allogeneic kidney transplants were performed between WF (RT1u) donors and Lewis (RT1) recipients. Donor kidneys were perfused in situ with saline alone (control), or 9 x 10(9) plaque-forming units of AdV-FasL. One native kidney was removed at the time of transplant and the other at 6 or 7 days. Uremic death was the endpoint, and deaths within 7 days of transplant were excluded. Transduced allografts were stained for FasL expression using a monoclonal antibody and tested for FasL mRNA production by reverse transcriptase-polymerase chain reaction and Northern blotting.
Immunostaining of AdV-FasL-transduced allografts demonstrated efficient gene transfer lasting approximately 2 weeks, and FasL mRNA production in the AdV-FasL-transduced allografts was confirmed by Northern blotting and reverse transcriptase-polymerase chain reaction. Mean survival of animals with AdV-FasL-transduced renal allografts was 27.8 days vs. 11.6 days in control animals (P < 0.05).
(1) Adenoviral vectors can successfully transduce rat kidneys with the FasL cDNA. (2) FasL gene transfer prolongs rat renal allograft survival.
Fas配体（FasL）可诱导表达其受体Fas的细胞发生凋亡，并且已证明其在T细胞发育与调节以及免疫赦免中发挥重要作用。我们推测肾移植受者中FasL的表达可能对移植排斥具有保护作用。
将小鼠FasL cDNA克隆到复制缺陷型腺病毒（AdV-FasL）中。通过对AdV-FasL转导的HeLa细胞进行免疫染色来确认蛋白表达。在WF（RT1u）供体和Lewis（RT1）受体之间进行同种异体肾移植。供体肾脏原位灌注单独的生理盐水（对照）或9×10⁹ 空斑形成单位的AdV-FasL。移植时切除一个自体肾，另一个在6或7天时切除。以尿毒症死亡作为观察终点，排除移植后7天内的死亡病例。使用单克隆抗体对转导的移植肾进行FasL表达染色，并通过逆转录-聚合酶链反应和Northern印迹法检测FasL mRNA的产生。
对AdV-FasL转导的移植肾进行免疫染色显示基因有效转移持续约2周，并且通过Northern印迹法和逆转录-聚合酶链反应确认了AdV-FasL转导的移植肾中FasL mRNA的产生。AdV-FasL转导的肾移植受者的平均生存期为27.8天，而对照动物为11.6天（P<0.05）。
（1）腺病毒载体可成功地将FasL cDNA转导至大鼠肾脏。（2）FasL基因转移可延长大鼠肾移植的存活时间。
