Institute of Experimental Oncology and Therapy Research, Klinikum rechts der Isar der Technischen Universität München, Munich, Germany.
Pharm Res. 2012 May;29(5):1219-31. doi: 10.1007/s11095-011-0631-2. Epub 2011 Dec 30.
To target adenoviral vectors to cells of the vasculature and shielding vectors from inactivation by the immune system.
Complexes of reporter gene expressing adenoviral vectors with positively charged magnetic nanoparticles were formed by electrostatic interaction in presence or absence of additional negatively charged poly(ethylene glycol)-based polymer. Transduction of HUVEC was analyzed in vitro under flow. Protection from inactivation by the immune system was analyzed by pre-incubation of AdV and complexes with neutralizing antibodies and subsequent reporter protein analysis of infected cells.
Physical association of AdV with MNP and polymers was demonstrated by radioactive labelling of components and co-sedimentation in a magnetic field. Ad-MNP+/-polymer resulted in efficient transduction of HUVEC, depending on MOI and flow rate in presence of magnetic field, whereas no transduction was observed without complex formation with MNP or in absence of magnetic field. Association with MNP did result in protection from neutralizing antibodies, with slightly increased protection provided by the polymer.
Complex formation of AdV with MNP is a viable means for targeting of vectors to areas of magnetic field gradient. Additional coating with polymer might proof useful in protection from inactivation by the immune system.
将腺病毒载体靶向血管细胞，并使载体免受免疫系统的失活。
通过静电相互作用形成表达报告基因的腺病毒载体与带正电荷的磁性纳米颗粒的复合物，存在或不存在额外的带负电荷的基于聚乙二醇的聚合物。在流动条件下分析体外 HUVEC 的转导。通过用中和抗体预孵育 AdV 和复合物，并随后分析感染细胞中的报告蛋白，分析对免疫系统失活的保护作用。
通过放射性标记成分和在磁场中共沉淀证明了 AdV 与 MNP 和聚合物的物理结合。Ad-MNP+/-聚合物在磁场存在下，取决于 MOI 和流速，可有效转导 HUVEC，而在没有与 MNP 形成复合物或没有磁场的情况下，则观察不到转导。与 MNP 的结合确实导致了对中和抗体的保护，聚合物提供了略微增加的保护。
AdV 与 MNP 的复合物形成是将载体靶向磁场梯度区域的一种可行方法。额外的聚合物涂层可能有助于防止免疫系统失活。
