Institute of Tropical Medicine of Antwerp, Nationalestraat 155, Antwerp 2000, Belgium.
Nanomedicine (Lond). 2013 Jan;8(1):77-87. doi: 10.2217/nnm.12.97. Epub 2012 Aug 14.
Cationic lipids (Lipofectamine™ [Invitrogen, Merelbeke, Belgium] and 1,2-dioleoyl-3-trimethylammonium-propane/1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) and polymers (jetPEI™ and in vivo-jetPEI™ [Polyplus-transfection, Illkirch, France]) were evaluated for their potential to deliver mRNA to monocyte-derived dendritic cells.
MATERIALS & METHODS: Lipoplexes and polyplexes, containing mRNA encoding GFP or Gag protein, were incubated with human monocyte-derived dendritic cells and transfection efficiencies were assessed by flow cytometry.
Lipofectamine was by far the most efficient in mRNA delivery, therefore it was used in further experiments. Incubation of monocyte-derived dendritic cells isolated from HIV-1-positive donors with mRNA encoding Gag protein complexed to Lipofectamine resulted in 50% transfection. Importantly, coculture of these Gag-transfected dendritic cells with autologous T cells induced an over tenfold expansion of IFN-γ- and IL-2-secreting CD4(+) and CD8(+) T cells.
Cationic lipid-mediated mRNA delivery may be a useful tool for therapeutic vaccination against HIV-1. This approach can be applied to develop vaccination strategies for other infectious diseases and cancer.
评估阳离子脂质（Lipofectamine™ [Invitrogen，比利时梅勒贝克]和 1,2-二油酰基-3-三甲铵丙烷/1,2-二油酰基-sn-甘油-3-磷酸乙醇胺）和聚合物（jetPEI™ 和 in vivo-jetPEI™ [Polyplus-transfection，法国伊尔基希]）将 mRNA 递送至单核细胞来源的树突状细胞的潜力。
用含有 GFP 或 Gag 蛋白编码 mRNA 的脂质体和聚合物与人类单核细胞来源的树突状细胞孵育，并通过流式细胞术评估转染效率。
Lipofectamine 是迄今为止最有效的 mRNA 递送方法，因此在进一步的实验中使用了它。与 Lipofectamine 复合的编码 Gag 蛋白的 mRNA 孵育分离自 HIV-1 阳性供体的单核细胞来源的树突状细胞导致 50%的转染。重要的是，将这些 Gag 转染的树突状细胞与自体 T 细胞共培养诱导 IFN-γ 和 IL-2 分泌的 CD4(+)和 CD8(+)T 细胞扩增超过十倍。
阳离子脂质介导的 mRNA 递呈可能是针对 HIV-1 的治疗性疫苗接种的有用工具。这种方法可用于开发针对其他传染病和癌症的疫苗接种策略。
