Genetic modification of murine dendritic cells by RNA transfection.
The ability to manipulate in vitro cultured dendritic cells (DCs) by transfection represents an attractive strategy to load these antigen-presenting cells with genetic material encoding various immunogenic epitopes. The gene transfer approach can also be applied to DCs with the aim of expressing immunologically active molecules such as cytokines, costimulatory molecules, or simply to transiently express proteins to perform cell biology studies. Available gene transfer technologies for DCs include both viral and non-viral vector-based approaches. In this chapter, we describe non-viral strategies of RNA transfection. Special emphasis is given to murine bone-marrow-derived DCs, since gene transfer to human DCs has been extensively described in the literature, especially in the context of cancer immunotherapy and other clinical applications. Methods to deliver small interfering RNA (siRNA) to DCs are described as well. Finally, the potential of exogenously delivered RNA to activate DCs is discussed and some practical advice to avoid DC activation is described.
Centre d'Immunologie de Marseille-Luminy, Université de la Méditerranée, Case 906, 13288, INSERM U631, CNRS, UMR6102, Marseille, France. ceppi@ciml.univ-mrs.fr
Maurizio
M
Ceppi
Enrico
E
Schmidt
Philippe
P
Pierre
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D018274:Electroporation; D016130:Immunophenotyping; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D012313:RNA / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection / Q000379:methods*
D008070:Lipopolysaccharides; D012333:RNA, Messenger; D012313:RNA