Benteyn Daphné, Van Nuffel An M T, Wilgenhof Sofie, Bonehill Aude
Laboratory of Molecular and Cellular Therapy, Department of Immunology-Physiology and the Dendritic Cell Bank, Vrije Universiteit Brussel, Brussels, Belgium.
Methods Mol Biol. 2014;1139:3-15. doi: 10.1007/978-1-4939-0345-0_1.
Dendritic cells (DC) are key players in several types of cancer vaccines. Large numbers of DC can easily be generated in closed systems from the monocyte fraction of the peripheral blood. They are the professional antigen-presenting cells, and electroporation of mRNA-encoding tumor antigens is a very efficient and a relatively simple way to load the DC with antigen. The co-electroporation of a tumor antigen of choice and the combination of 3 costimulatory molecules, including CD70, caTLR4, and CD40L (TriMix-DC), leads to fully potent antigen-presenting DC able to generate a broad immune response.Here we describe the in vitro transcription of the mRNA and the subsequent generation and electroporation of autologous DC used for the treatment of melanoma patients.
树突状细胞（DC）是几种癌症疫苗中的关键成分。在外周血单核细胞部分的封闭系统中可以轻松大量生成DC。它们是专职抗原呈递细胞，对编码肿瘤抗原的mRNA进行电穿孔是一种非常有效且相对简单的用抗原加载DC的方法。选择的肿瘤抗原与包括CD70、caTLR4和CD40L在内的3种共刺激分子（TriMix-DC）联合电穿孔，可产生完全有效的抗原呈递DC，能够引发广泛的免疫反应。在此，我们描述用于治疗黑色素瘤患者的mRNA的体外转录以及随后自体DC的生成和电穿孔。
