Ali Omar A, Mooney David J
School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, USA.
J Control Release. 2008 Dec 18;132(3):273-8. doi: 10.1016/j.jconrel.2008.07.005. Epub 2008 Jul 10.
Current techniques to educate dendritic cells (DCs) ex vivo for immunotherapy are plagued by inefficient protocols and DC modifications are often transient and lost upon transplantation. This study investigated the role of sustained presentation of GM-CSF and PEI condensed pDNA (PEI-DNA) on gene transfer and long-term gene expression. Appropriate GM-CSF signaling during DC transfection promoted PEI-DNA uptake, although high cytokine concentrations induced intercellular DNA degradation, indicating the need for controlled presentation. Poly(lactide-co-glycolide) scaffolds that continuously stimulated DCs with both GM-CSF and PEI-DNA led to a 20-fold increase in gene expression, and high levels of expression persisted for a period of 10 days, in vitro. These results encourage the exploitation of biomaterials and GM-CSF to develop novel delivery vectors for genetically modified DCs or to genetically program host DCs in situ for vaccination and the treatment of autoimmunity.
目前用于体外培养树突状细胞（DCs）以进行免疫治疗的技术存在方案效率低下的问题，而且DC修饰往往是短暂的，在移植后会消失。本研究调查了持续呈现粒细胞巨噬细胞集落刺激因子（GM-CSF）和聚乙烯亚胺（PEI）缩合的质粒DNA（PEI-DNA）对基因转移和长期基因表达的作用。DC转染过程中适当的GM-CSF信号传导促进了PEI-DNA的摄取，尽管高细胞因子浓度会诱导细胞内DNA降解，这表明需要进行可控的呈现。用GM-CSF和PEI-DNA持续刺激DCs的聚乳酸-羟基乙酸共聚物（PLGA）支架导致基因表达增加了20倍，并且在体外高水平表达持续了10天。这些结果鼓励利用生物材料和GM-CSF来开发用于基因修饰DCs的新型递送载体，或对宿主DCs进行原位基因编程以用于疫苗接种和自身免疫性疾病的治疗。
