Lavigne C, Thierry A R
Département de Microbiologie et Immunologie, Faculté de Médecine, Université de Montréal, Québec, Canada.
Biochem Biophys Res Commun. 1997 Aug 28;237(3):566-71. doi: 10.1006/bbrc.1997.7191.
The relatively poor cell uptake of oligonucleotides and subsequent transport to the cytoplasm and nucleus is the main limitation in antisense therapeutics. The use of lipid-based carrier system is one of the most promising approaches to overcome these problems. In this study, we report the use of a new lipidic formulation to deliver a phosphorothioate oligonucleotide antisense directed against the regulatory gene rev of the HIV-1 genome and its application to the inhibition of HIV-1 in different cell culture models. Antiviral activity of either DLS-complexed or non-complexed oligonucleotides (ODNs) was compared in acutely and chronically infected cells. We have demonstrated that substantial antisense activity could be achieved at subnanomolar concentrations with DLS-complexed ODN in both acute and chronic infection systems. DLS-association highly improved inhibitory activity of the antisense ODN in acutely infected Molt-3 cells (100-fold) and primary cells (1000-fold) and in chronically infected H9 cells (1,500,000-fold). We have shown that anti-HIV activity of phosphorothioate ODNs can be strongly enhanced by using the DLS carrier system.
寡核苷酸相对较差的细胞摄取以及随后向细胞质和细胞核的转运是反义疗法的主要限制。使用基于脂质的载体系统是克服这些问题最有前景的方法之一。在本研究中，我们报告了一种新型脂质制剂用于递送针对HIV-1基因组调控基因rev的硫代磷酸酯寡核苷酸反义药物，及其在不同细胞培养模型中对HIV-1的抑制作用。在急性和慢性感染细胞中比较了与DLS复合或未复合的寡核苷酸（ODN）的抗病毒活性。我们已经证明，在急性和慢性感染系统中，与DLS复合的ODN在亚纳摩尔浓度下均可实现显著的反义活性。DLS结合显著提高了反义ODN在急性感染的Molt-3细胞（100倍）和原代细胞（1000倍）以及慢性感染的H9细胞（1500000倍）中的抑制活性。我们已经表明，使用DLS载体系统可以强烈增强硫代磷酸酯ODN的抗HIV活性。
