Development of adenovirus immobilization strategies for in situ gene therapy.
Development of adenovirus immobilization strategies for in situ gene therapy.
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DOI：
10.1002/jgm.1233
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发表时间：
2008-10
影响因子：
3.5
通讯作者：
Krebsbach, Paul H.
中科院分区：
文献类型：
--
作者：
Hu, Wei-Wen;Lang, Michael W.;Krebsbach, Paul H.
使用病毒载体的再生基因治疗使转导细胞能够在体内表达生物活性因子。空间控制的病毒传递可以提高转导效率，避免全身感染。因此，我们通过与壳聚糖表面的亲和素相互作用将生物素化的腺病毒捆绑在一起，以获得对原位转导的强大控制。将亲和素直接连接到壳聚糖(病毒-生物素-亲和素材料；VBAM)或间接对接在生物素化的壳聚糖表面(病毒-生物素-亲和素-生物素材料(VBABM))来锚定生物素化的腺病毒。用酶联免疫吸附试验(ELISA)和光谱分析验证了结合图谱。以生物素碱性磷酸酶和生物素化的腺病毒作为不同大小的粒子，评价其结合效率，并用SIPS等温吸附法进行比较。扫描电子显微镜(SEM)显示病毒的分布，通过体外细胞转导来确定转导效率。酶联免疫吸附试验和光谱分析均表明，VBAM体系在生物材料表面形成多层亲和素，而VBABM形成单层亲和素。SIPS等温线吸附表明，VBAM方法增加了结合位的不均一性和空间位阻。相比之下，VBABM方法将亲和素对接在壳聚糖表面，并定位结合部位以促进配体结合。此外，扫描电子显微镜图像显示，VBABM方法导致了更均匀的病毒分布。体外细胞感染实验还表明，VBABM系统增强了病毒的固定化，从而提高了细胞转导效率。VBABM策略是一种从生物材料中进行原位转导的优越方法。这一策略可以适用于各种生物材料和病毒载体，因此可能成为体内再生基因治疗的一种替代方法。
Regenerative gene therapy using viral vectors enables transduced cells to express bioactive factors in vivo. Viral delivery with spatial control can enhance transduction efficiency and avoid systemic infection. Consequently, we tethered biotinylated adenovirus via interactions with avidin on chitosan surfaces to gain robust control for in situ transduction. Avidin was either directly conjugated to chitosan (virus-biotin-avidin material; VBAM) or indirectly docked on biotinylated chitosan surfaces (virus-biotin-avidin-biotin material; VBABM) to tether biotinylated adenovirus. Enzyme-linked immunosorbent assay (ELISA) and spectroscopic analysis were performed to demonstrate the binding profiles. Biotin-alkaline phosphatase and biotinylated adenovirus were used as different sized particles to evaluate binding efficiencies and were compared by the Sips isotherm adsorption method. Scanning electron microscopy (SEM) examination illustrated virus distribution, and the transduction efficiency was determined by in vitro cell transduction. ELISA and spectroscopic analysis both demonstrated that the VBAM system led to multilayer avidin formation on biomaterial surfaces, whereas VBABM formed a monolayer of avidin. Sips isotherm adsorption indicated that the VBAM method increased heterogeneity and steric hindrance of binding sites. By contrast, the VBABM method docked avidin on chitosan surfaces and orientated the binding sites to facilitate ligand binding. In addition, SEM images illustrated that the VBABM method led to more even viral distribution. In vitro cell infection experiments also revealed that the VBABM system enhanced virus immobilization and thus improved cell transduction efficiency over the VBAM system. The VBABM strategy is a superior method for in situ transduction from biomaterials. This strategy could be adapted for use with a variety of biomaterials as well as viral vectors, and thus may be an alternative method for in vivo regenerative gene therapy.
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影响因子：
4.2
作者：
Klugherz, BD;Song, CX;Levy, RJ
通讯作者：
Levy, RJ
影响因子：
3.5
作者：
Bengali, Zain;Rea, Jennifer C.;Shea, Lonnie D.
通讯作者：
Shea, Lonnie D.
影响因子：
3.5
作者：
Mei, Lin;Jin, Xu;Levy, R. J.
通讯作者：
Levy, R. J.
影响因子：
5.1
作者：
Levy, RJ;Song, C;Li, Q
通讯作者：
Li, Q
影响因子：
2.9
作者：
Bajaj, B;Lei, P;Andreadis, ST
通讯作者：
Andreadis, ST