Acute responses of non-human primates to airway delivery of an adenovirus vector containing the human cystic fibrosis transmembrane conductance regulator cDNA.
Acute responses of non-human primates to airway delivery of an adenovirus vector containing the human cystic fibrosis transmembrane conductance regulator cDNA.
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非人灵长类动物对含有人囊性纤维化跨膜电导调节剂 cDNA 的腺病毒载体气道递送的急性反应。
DOI：
10.1089/hum.1994.5.7-821
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发表时间：
1994
影响因子：
4.2
通讯作者：
Ronald G. Crystal
中科院分区：
文献类型：
--
作者：
Steven L. Brody;Mark Metzger;Claire Danel;M. Rosenfeld;Ronald G. Crystal
重组人腺病毒（Ad）载体是囊性纤维化（CF）的人基因治疗的主要候选者，其基于在体内将外源基因有效转移到啮齿动物呼吸上皮和在体外将外源基因转移到人呼吸细胞的证明。在恒河猴中评价了Ad介导的基因转移至呼吸道上皮的安全性和对含有人囊性纤维化跨膜传导调节因子cDNA（AdCFTR）的基于复制缺陷型重组E1-，E3- Ad 5型载体的气道递送的急性（长达21天）临床反应。气道递送具有lacZ标记基因的Ad载体证明了上皮细胞中的β-半乳糖苷酶表达。经气管内注射AdCFTR的动物在气道上皮细胞中显示人CFTR cDNA表达。鼻内给予AdCFTR的动物，以及24小时后，支气管内给予AdCFTR的动物[2 × 10（7）至5 × 10（10）空斑形成单位（pfu），n = 12]，以类似于拟定的人类方案的方式，或仅支气管内给予AdCFTR的动物（10（11）pfu，n = 3），与溶剂对照组（n = 6）相比，临床参数无显著变化。通过尸检或支气管肺泡灌洗对肺进行的显微镜分析表明，在AdCFTR递送的区域中，炎性细胞（主要是淋巴细胞）呈剂量依赖性增加，在一些动物中持续至少2个月。血清抗Ad 5型中和抗体滴度未升高，未检测到脱落Ad。通过聚合酶链反应（PCR）分析，在肺外器官中未检测到AdCFTR DNA的存在。这些数据表明，AdCFTR在非人灵长类动物中耐受良好，尽管在肺中存在临床上不明显的剂量依赖性炎症。（250字处删节）
Recombinant human adenovirus (Ad) vectors are leading candidates for human gene therapy for cystic fibrosis (CF) based on demonstration of efficient transfer of exogenous genes to rodent respiratory epithelium in vivo and human respiratory cells in vitro. The safety of Ad-mediated gene transfer to the respiratory epithelium and acute (up to 21 days) clinical responses to airway delivery of a replication-deficient recombinant, E1-, E3- Ad type 5-based vector containing the human cystic fibrosis transmembrane conductance regulator cDNA (AdCFTR) were evaluated in rhesus monkeys. Airway delivery of an Ad vector with the lacZ marker gene demonstrated beta-galactosidase expression in epithelial cells. Animals administered intratracheal AdCFTR demonstrated human CFTR cDNA expression in airway epithelial cells. Animals administered AdCFTR intranasal, and 24 hr later, intrabronchial [2 x 10(7) to 5 x 10(10) plaque-forming units (pfu), n = 12], in a fashion similar to a proposed human protocol, or only intrabronchial (10(11) pfu, n = 3), had no significant changes in clinical parameters compared to vehicle controls (n = 6). Microscopic analysis of the lung by necropsy or bronchoalveolar lavage demonstrated a dose-dependent increase in inflammatory cells, primarily lymphocytes, in the area where AdCFTR was delivered, which persisted for at least 2 months in some animals. Serum anti-Ad type 5 neutralizing antibody titers did not rise and shed Ad was not detected. The presence of AdCFTR DNA, analyzed by the polymerase chain reaction (PCR), was not detected in organs outside the lung. These data demonstrate that AdCFTR is well tolerated in non-human primates, although there is dose-dependent inflammation in the lung not clinically apparent.(ABSTRACT TRUNCATED AT 250 WORDS)