Simon R H, Engelhardt J F, Yang Y, Zepeda M, Weber-Pendleton S, Grossman M, Wilson J M
Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor 48109.
Hum Gene Ther. 1993 Dec;4(6):771-80. doi: 10.1089/hum.1993.4.6-771.
In preparation for human trials of gene therapy for cystic fibrosis (CF), we performed a preclinical study of gene transfer into the lungs of baboons. Recombinant adenovirus vectors containing expression cassettes for human cystic fibrosis transmembrane conductance regulator (CFTR) and Escherichia coli beta-galactosidase (lacZ) were instilled through a bronchoscope into limited regions of lung in 14 baboons. A detailed accounting of the extent, distribution, and duration of gene expression is contained in a companion article (Engelhardt et al., 1993b). In this article, we report the results of toxicity studies in which clinical laboratory tests, chest radiographs, and necropsy studies were used to detect adverse effects. The only adverse effect noted was a mononuclear cell inflammatory response within the alveolar compartment of animals receiving doses of virus that were required to induce detectable gene expression. Minimal inflammation was seen at 10(7) and 10(8) pfu/ml, but at 10(9) and more prominently at 10(10) pfu/ml, a perivascular lymphocytic and histiocytic infiltrate was seen. The intensity of inflammation increased between 4 and 21 days. At its greatest intensity, there was diffuse alveolar wall damage with intra-alveolar edema. Airways were relatively spared, despite the intensity of alveolar inflammation. Clinical tests did not accurately reflect the presence of lung inflammation, with the exception of chest radiographs which revealed alveolar infiltrates, but only in regions of lung having the greatest intensity inflammation. We conclude that adenovirus-mediated gene transfer into the lungs of baboons is associated with development of alveolar inflammation at high doses of virus.
为了准备针对囊性纤维化（CF）的基因治疗进行人体试验，我们对狒狒肺部的基因转移进行了一项临床前研究。将含有人类囊性纤维化跨膜传导调节因子（CFTR）和大肠杆菌β-半乳糖苷酶（lacZ）表达盒的重组腺病毒载体通过支气管镜注入14只狒狒肺部的有限区域。基因表达的范围、分布和持续时间的详细情况包含在一篇配套文章中（Engelhardt等人，1993b）。在本文中，我们报告了毒性研究的结果，其中使用临床实验室检查、胸部X光片和尸检研究来检测不良反应。唯一观察到的不良反应是在接受诱导可检测基因表达所需病毒剂量的动物的肺泡隔内出现单核细胞炎症反应。在10^7和10^8 pfu/ml时可见轻微炎症，但在10^9 pfu/ml时，尤其是在10^10 pfu/ml时，可见血管周围淋巴细胞和组织细胞浸润。炎症强度在4至21天之间增加。在炎症最严重时，出现弥漫性肺泡壁损伤伴肺泡内水肿。尽管肺泡炎症严重，但气道相对未受影响。除了胸部X光片显示肺泡浸润外，临床检查并不能准确反映肺部炎症的存在，而且只有在炎症最严重的肺部区域才会出现肺泡浸润。我们得出结论，高剂量病毒介导的腺病毒基因转移到狒狒肺部与肺泡炎症的发生有关。
