Pickles R J, McCarty D, Matsui H, Hart P J, Randell S H, Boucher R C
CF/Pulmonary Research and Treatment Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7248, USA.
J Virol. 1998 Jul;72(7):6014-23. doi: 10.1128/JVI.72.7.6014-6023.1998.
Investigations of the efficiency and safety of human adenovirus vector (AdV)-mediated gene transfer in the airways of patients with cystic fibrosis (CF) in vivo have demonstrated little success in correcting the CF bioelectrical functional defect, reflecting the inefficiency of AdV-mediated gene transfer to the epithelial cells that line the airway luminal surface. In this study, we demonstrate that low AdV-mediated gene transfer efficiency to well-differentiated (WD) cultured airway epithelial cells is due to three distinct steps in the apical membrane of the airway epithelial cells: (i) the absence of specific adenovirus fiber-knob protein attachment receptors; (ii) the absence of alphavbeta3/5 integrins, reported to partially mediate the internalization of AdV into the cell cytoplasm; and (iii) the low rate of apical plasma membrane uptake pathways of WD airway epithelial cells. Attempts to increase gene transfer efficiency by increasing nonspecific attachment of AdV were unsuccessful, reflecting the inability of the attached vector to enter (penetrate) WD cells via nonspecific entry paths. Strategies to improve the efficiency of AdV for the treatment of CF lung disease will require methods to increase the attachment of AdV to and promote its internalization into the WD respiratory epithelium.
对人类腺病毒载体（AdV）介导的基因转移在囊性纤维化（CF）患者气道内的体内效率和安全性的研究表明，在纠正CF生物电功能缺陷方面成效甚微，这反映出AdV介导的基因转移至气道管腔表面衬里上皮细胞的效率低下。在本研究中，我们证明AdV介导的基因向分化良好（WD）的培养气道上皮细胞的转移效率低是由于气道上皮细胞顶端膜上的三个不同步骤：（i）缺乏特异性腺病毒纤维-钮蛋白附着受体；（ii）缺乏αvβ3/5整合素，据报道其可部分介导AdV内化进入细胞质；以及（iii）WD气道上皮细胞顶端质膜摄取途径的速率较低。通过增加AdV的非特异性附着来提高基因转移效率的尝试未成功，这反映出附着的载体无法通过非特异性进入途径进入（穿透）WD细胞。提高AdV治疗CF肺部疾病效率的策略将需要增加AdV与WD呼吸道上皮的附着并促进其内化的方法。
