Localized adenovirus gene delivery using antiviral IgG complexation
Localized adenovirus gene delivery using antiviral IgG complexation
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DOI：
10.1038/sj.gt.3301452
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发表时间：
2001-05-01
期刊：
影响因子：
5.1
通讯作者：
Li, Q
中科院分区：
文献类型：
--
作者：
Levy, RJ;Song, C;Li, Q
利用病毒载体的基因治疗已经发展到临床试验阶段。然而，将病毒载体递送至患病靶位点的定位仍然是一个挑战。我们测试的假设，腺病毒载体可以成功地通过与一个特定的抗体，结合到一个生物可降解的矩阵设计实现本地化的基因转导复合交付。我们报告了第一个成功的递送系统的基础上抗体固定的病毒粒子在I型胶原蛋白-亲和素凝胶使用多克隆生物素化的特异性的腺病毒六邻体的IgG。体外稳定性研究表明，与用非特异性生物素化IgG制备的胶原凝胶的载体活性损失相比，用抗体复合的腺病毒胶原凝胶制剂保留了病毒载体活性。使用这种抗体控制释放系统进行大鼠主动脉平滑肌细胞（A10）的腺病毒载体转导的细胞培养研究表明，与非抗体复合对照相比，报告基因表达（β-半乳糖苷酶）明显更局限。将单纯疱疹胸苷激酶（HSVtk）腺病毒载体通过这种抗体复合方法固定在亲和素-胶原凝胶上，并将更昔洛韦加入到与凝胶一起培养的大鼠平滑肌细胞（A10）中。用复合的HSVtk腺病毒，只有与含病毒凝胶接触或在50 μ m内的细胞被杀死。相比之下，在相同的腺病毒和更昔洛韦剂量下，用更昔洛韦递送非抗体复合的HSVtk腺病毒导致几乎所有细胞死亡。猪心肌基因转移研究表明，与直接载体注射相比，抗六邻体抗体复合基质注射显著更有效的右心室腺病毒GFP表达。因此，我们的研究结果表明，基于腺病毒的抗体复合递送的基质制剂导致转基因表达的位点特异性定位，这增强了治疗载体策略的效率，并提供了有效的定位手段，以避免远端副作用。这种方法具有作为植入式制剂的治疗潜力，通过抗体复合的手段，可以定位和优化病毒载体基因治疗。
Gene therapy with viral vectors has progressed to clinical trials. However, the localization of viral vector delivery to diseased target sites remains a challenge. We tested the hypothesis that an adenoviral vector could be successfully delivered by complexation with a specific antibody that is bound to a biodegradable matrix designed for achieving localized gene transduction. We report the first successful delivery system based upon antibody immobilization of virions in a type I collagen-avidin gel using a polyclonal biotinylated IgG specific for the adenovirus hexon. In vitro stability studies demonstrated retention of viral vector activity with antibody-complexed adenovirus collagen gel preparations, in comparison to loss of vector activity from collagen gels prepared with nonspecific biotinylated IgG. Cell culture investigations using this antibody-controlled release system for adenoviral vector transduction of rat aortic smooth muscle cells (A10) demonstrated a significantly more localized reporter expression (P-galactosidase) compared with non-antibody-complexed controls. Herpes simplex thymidine kinase (HSVtk) adenoviral vectors were immobilized on avidin-collagen gels via this antibody-complexation approach, and ganciclovir was added to rat smooth muscle cells (A10) in culture with the gels. With complexed HSVtk adenovirus, only cells either in contact with the virus-containing gel or within 50 mum were killed By comparison, at the same adenovirus and ganciclovir dose, non-antibody-complexed HSVtk adenoviral delivery with ganciclovir resulted in the death of virtually all cells. Myocardial gene transfer studies in pigs demonstrated significantly more efficient right ventricular adenoviral GFP expression with anti-hexon antibody-complexed matrix injections, compared with direct Vector injections. Thus, our results show that matrix formulations based on antibody-complexation delivery of adenovirus resulted in site-specific localization of transgene expression that enhances the efficiency of therapeutic vector strategies and provides a potent means for localization, to avoid distal side-effects. This approach has therapeutic potential as an implantable preparation that through the means of antibody-complexation, can localize and optimize viral vector gene therapy.