摘要
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。
We report a successful delivery system based upon antibody immobilization of virions in a type Ⅰ collagen-avidin gel using polyclonal biotinylated IgG specfic 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 from nonspecific biotinylated IgG. Herpes simplex thymidine kinase （HSVtk） adenovirus vectors were immobilized on avidin-collagen gels via this antibody-complexation approach, and ganciclovir was added to rat aortic 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 the range of 50μm around film were killed. By comparison, at the same adenovirus and ganciclovir dose, non-antibody-complexed HSVtk adenovirus delivery with ganciclovir resulted in the death of virtually all cells. Myocardial gene transfer studied in pigs demonstrated significantly more efficient right ventricular adenovirus GFP expression with anti-hexon antibody-complexed matrix injections, compared with direct vector injections. The results show that matrix formulation based on antibody-complexation delivery of adenovirus resulted in site-specific localization of transgene expression that enhanced the efficiency of therapeutic potential as an implantable preparation by means of antibody-complexation. The method provided a new way of localizing and optimizing viral vector gene therapy.
出处
《中国生物医学工程学报》
CAS
CSCD
北大核心
2005年第5期590-595,共6页
Chinese Journal of Biomedical Engineering
基金
国家自然基金项目50473059
博士点专项基金20030023004
天津市应用基础研究重点项目04380311。
关键词
靶向特异
定位递送
抗体偶联
腺病毒载体
site-specific
local delivery
antibody-mediated
adenoviral vector
