Delivery of tumor-antigen-encoding mRNA into dendritic cells for vaccination.
Antigen-loaded dendritic cells (DCs) have been intensively investigated as potential cellular antitumor vaccines. Several recent reports have indicated that loading DCs with whole tumor derived mRNA or defined tumor-antigen-encoding mRNA represents an effective nonviral strategy to stimulate T cell responses both for in vitro and in vivo models. Here, we describe the electroporation method as a tool for introducing in vitro transcribed capped mRNA into human DCs for tumor vaccination. We use MART-1/Melan-A as a model tumor-associated antigen for the generation of a DC-based vaccine against melanoma cancer. In addition to efficient antigen loading, it is important to obtain a maximal number of potent antigen-presenting cells. Another prerequisite for the development of a DC-based cancer vaccine is to obtain mature DCs. In this chapter, we describe the basic techniques required for the successful genetic modification of DCs by using the mRNA electroporation method.
Michiels
Annelies
A
Department of Physiology and Immunology, Medical School of the Vrije Universiteit Brussel , Brussels, Belgium.
Tuyaerts
Sandra
S
Bonehill
Aude
A
Heirman
Carlo
C
Corthals
Jurgen
J
Thielemans
Kris
K
D000951:Antigens, Neoplasm / Q000235:genetics*; D001483:Base Sequence; D019496:Cancer Vaccines / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D002469:Cell Separation; D017931:DNA Primers / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology*; D053672:Electrochemotherapy / Q000379:methods; D018274:Electroporation / Q000379:methods; D006801:Humans; D058965:MART-1 Antigen; D008545:Melanoma / Q000235:genetics / Q000276:immunology / Q000628:therapy; D016297:Mutagenesis, Site-Directed; D009363:Neoplasm Proteins / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D012315:RNA Caps / Q000235:genetics; D012333:RNA, Messenger / Q000008:administration & dosage* / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology
D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D017931:DNA Primers; D058965:MART-1 Antigen; C548667:MLANA protein, human; D009363:Neoplasm Proteins; D012315:RNA Caps; D012333:RNA, Messenger