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临床基因治疗方案中基因载体在疾病部位的靶向递送仍是急待解决的问题。本研究将腺病毒与一种生物素化的特异性抗腺病毒六邻体的多克隆IgG结合,固定于结合了亲和素的胶原蛋白膜上,成功获得腺病毒载体基因靶向定位递送体系。体外稳定性研究结果表明该体系中病毒载体可有效保持活性。通过这种特异性抗体偶联方式,将携带单纯疱疹胸苷激酶(HSVtk)编码基因片断的腺病毒结合在胶原膜上转染大鼠平滑肌细胞(A10),加入更昔洛韦(ganciclovir)后,只有生长在胶原膜上及膜邻近50μm内的细胞被杀死。在使用非特异性抗体的对照实验中,整个培养基范围内的细胞几乎全被杀死。以绿色荧光蛋白(GFP)为报告基因对猪的心肌进行转基因实验,结果显示注射特异性抗体偶联病毒的胶原凝胶比直接注射病毒悬液获得更高效的心室基因表达。所有研究结果表明,通过生物素和特异性抗体使病毒载体固定在胶原蛋白基质上,可达到有效的局部定位基因表达,避免向非病灶部位的扩散,是基因治疗中一种极具发展潜力的载体定位递送方法。
Targeted delivery of gene vectors at the site of disease in clinical gene therapy regimens remains a pressing issue. In this study, adenovirus was combined with polyclonal IgG, a biotinylated specific anti-adenovirus hexon, and immobilized on an avidin-conjugated collagen membrane to successfully obtain the adenoviral vector gene targeting and localization delivery system . In vitro stability studies show that the virus vector in this system can effectively maintain the activity. By this specific antibody conjugation method, the adenoviruses carrying herpes simplex virus thymidine kinase (HSVtk) -encoding gene fragments are bound to the collagen membrane and transfected into rat smooth muscle cells (A10). After addition of ganciclovir Only cells that grew on the collagen membrane and within 50 μm of the membrane were killed. In control experiments using non-specific antibodies, almost all of the cells in the entire culture medium were killed. Transgenic experiments of porcine myocardium with green fluorescent protein (GFP) reporter gene showed that collagen gel injected with specific antibody-conjugated virus obtained more efficient ventricular gene expression than direct injection of virus suspension. All the results show that biotin and specific antibodies to the viral vector immobilized on the collagen matrix, can achieve effective localization of gene expression, to avoid the spread to non-focal sites, is a gene therapy with great potential Vector Positioning Delivery Methods.