Transduction by adeno-associated virus vectors in the rabbit airway: Efficiency, persistence, and readministration
Transduction by adeno-associated virus vectors in the rabbit airway: Efficiency, persistence, and readministration
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DOI：
10.1128/jvi.71.8.5932-5941.1997
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发表时间：
1997-08-01
影响因子：
5.4
通讯作者：
Miller, AD
中科院分区：
文献类型：
--
作者：
Halbert, CL;Standaert, TA;Miller, AD
重组腺相关病毒（AVV）载体整合宿主基因组和转导非分裂细胞的能力使其成为基因传递的载体。在这项研究中，我们通过测量标记基因的表达来评估几种AAV载体转导家兔气道细胞的能力。利用在透视引导下放置的球囊导管将转移β -半乳糖苷酶（β -gal）或人胎盘碱性磷酸酶（AP）基因的AAV载体传递到兔肺的一个肺叶，我们观察到在球囊放置区域的支气管上皮和平滑肌中几乎只存在载体编码的β -gal或AP染色。球囊处理支气管上皮的整体转导效率较低，但在某些区域可达20%。我们观察到，在载体输注后的60天内，标记阳性的上皮细胞减少了80倍，而标记阳性的平滑肌细胞的数量保持不变。虽然用拓扑异构酶抑制剂etoposide治疗显著增强了AAV在培养的原代气道上皮细胞中的转导，但家兔的处理并没有提高气道中的转导率，载体再给药未能产生额外的转导事件，这与中和抗体的出现有关。这些结果表明，在使用AAV载体对气道上皮进行基因治疗时，需要重新给药和免疫调节。
The ability of recombinant adeno-associated virus (AVV) vectors to integrate into the host genome and to transduce nondividing cells makes them attractive as vehicles for gene delivery. In this study, we assessed the ability of several AAV vectors to transduce airway cells in rabbits by measuring marker gene expression. AAV vectors that transferred either a beta-galactosidase (beta-gal) or a human placental alkaline phosphatase (AP) gene were delivered to one lobe of the rabbit lung by use of a balloon catheter placed under fluoroscopic guidance, We observed vector-encoded beta-gal or AP staining almost exclusively in the epithelial and smooth muscle tells in the bronchus at the region of balloon placement, The overall efficiency of transduction in the balloon-treated bronchial epithelium was low but reached 20% in some areas. The majority of the staining was in ciliated cells but was also observed in basal cells and airway smooth muscle cells, We observed an 80-fold decrease in marker-positive epithelial cells during the 60-day period after vector infusion, whereas the number of marker-positive smooth muscle cells stayed constant. Although treatment with the topoisomerase inhibitor etoposide dramatically enhanced AAV transduction in primary airway epithelial cells in culture, treatment of rabbits did not improve transduction rates in the airway, Vector readministration failed to produce additional transduction events, which correlated with the appearance of neutralizing antibodies. These results indicate that both readministration and immune modulation will be required in the use of AAV vectors for gene therapy to the airway epithelium.