Wirtz S, Galle P R, Neurath M F
Laboratory of Immunology, I Medical Clinic, University of Mainz, Langenbeckstrasse, 55101 Mainz, Germany.
Gut. 1999 Jun;44(6):800-7. doi: 10.1136/gut.44.6.800.
BACKGROUND/AIMS: Replication deficient recombinant adenoviruses represent an efficient means of transferring genes in vivo into a wide variety of dividing and quiescent cells from many different organs. Although the gastrointestinal tract is a potentially attractive target for gene therapy approaches, only a few studies on the use of viral gene transfer vehicles in the gut have been reported. The prospects of using recombinant adenoviruses for gene delivery into epithelial and subepithelial cells of the normal and inflamed colon are here analysed.
An E1/E3 deleted recombinant adenovirus (denoted AdCMVbetaGal) and an adenovirus with modified fibre structure (denoted AdZ.F(pk7)) both expressing the bacterial lacZ gene under the control of a human cytomegalovirus promoter were used for reporter gene expression in vitro and in vivo. beta-Galactosidase activity was determined by specific chemiluminescent reporter gene assay.
Intravenous or intraperitoneal injection of AdCMVbetaGal into healthy Balb/c mice caused strong reporter gene expression in the liver and spleen but not in the colon. In contrast, local administration of AdCMVbetaGal resulted in high reporter gene expression in colonic epithelial cells and lamina propria mononuclear cells. A local route of adenovirus administration in mice with experimental colitis induced by the hapten reagent trinitrobenzenesulphonic acid was next evaluated. Interestingly, rectal administration of AdCMVbetaGal caused a higher beta-galactosidase activity in isolated lamina propria cells from infected mice with experimental colitis than in those from controls. Furthermore, isolated lamina propria cells from mice with colitis infected in vitro showed a significant increase in reporter gene activity compared with controls. Finally, AdZ.F(pk7) adenoviruses with modified fibre structure produced 10- to 40-fold higher reporter gene activity in spleen T cells and lamina propria mononuclear cells of colitic mice compared with standard AdCMVbetaGal vectors.
Local administration of recombinant adenoviruses with normal or modified fibre structure could provide a new reliable method for targeted gene expression in the inflamed colon. Such gene delivery could be used to specifically express signal transduction proteins with therapeutic potential in inflamed colonic tissue. In particular, adenoviruses with modified fibre structure may be useful in T cell directed therapies in intestinal inflammation.
背景/目的：复制缺陷型重组腺病毒是一种将基因体内转移至来自许多不同器官的多种分裂和静止细胞的有效手段。尽管胃肠道是基因治疗方法潜在的有吸引力的靶点，但关于在肠道中使用病毒基因转移载体的研究报道较少。本文分析了使用重组腺病毒将基因递送至正常和炎症结肠的上皮及上皮下细胞的前景。
一种E1/E3缺失的重组腺病毒（命名为AdCMVbetaGal）和一种具有修饰纤维结构的腺病毒（命名为AdZ.F(pk7)），二者均在人巨细胞病毒启动子控制下表达细菌lacZ基因，用于体外和体内的报告基因表达。通过特异性化学发光报告基因测定法测定β-半乳糖苷酶活性。
将AdCMVbetaGal静脉内或腹腔内注射到健康的Balb/c小鼠中，导致肝脏和脾脏中报告基因强烈表达，但结肠中无表达。相反，局部给予AdCMVbetaGal导致结肠上皮细胞和固有层单核细胞中报告基因高表达。接下来评估了在由半抗原试剂三硝基苯磺酸诱导的实验性结肠炎小鼠中腺病毒的局部给药途径。有趣的是，直肠给予AdCMVbetaGal导致来自感染实验性结肠炎小鼠的分离固有层细胞中的β-半乳糖苷酶活性高于对照组。此外，与对照组相比，体外感染的结肠炎小鼠分离的固有层细胞中报告基因活性显著增加。最后，与标准AdCMVbetaGal载体相比，具有修饰纤维结构的AdZ.F(pk7)腺病毒在结肠炎小鼠的脾脏T细胞和固有层单核细胞中产生的报告基因活性高10至40倍。
局部给予具有正常或修饰纤维结构的重组腺病毒可为炎症结肠中的靶向基因表达提供一种新的可靠方法。这种基因递送可用于在炎症结肠组织中特异性表达具有治疗潜力的信号转导蛋白。特别是，具有修饰纤维结构的腺病毒可能在肠道炎症的T细胞定向治疗中有用。
