title: Electroporation of immature and mature dendritic cells: implications for dendritic cell-based vaccines.
abstract: Until now, studies utilizing mRNA electroporation as a tool for the delivery of tumor antigens to human monocyte-derived dendritic cells (DC) have focused on DC electroporated in an immature state. Immature DC are considered to be specialized in antigen capture and processing, whereas mature DC present antigen and have an increased T-cell stimulatory capacity. Therefore, the consensus has been to electroporate DC before maturation. We show that the transfection efficiency of DC electroporated either before or after maturation was similarly high. Both immature and mature electroporated DC, matured in the presence of an inflammatory cytokine cocktail, expressed mature DC surface markers and preserved their capacity to secrete cytokines and chemokines upon CD40 ligation. In addition, both immature and mature DC can be efficiently cryopreserved before or after electroporation without deleterious effects on viability, phenotype or T-cell stimulatory capacity including in vitro antigen-specific T-cell activation. However, DC electroporated after maturation are more efficient in in vitro migration assays and at least as effective in antigen presentation as DC electroporated before maturation. These results are important for vaccination strategies where an optimal antigen presentation by DC after migration to the lymphoid organs is crucial.
authors.affiliation: Laboratory of Molecular and Cellular Therapy, Department of Physiology-Immunology, Medical School of the Vrije Universiteit Brussel (VUB), Brussels, Belgium.
authors.forename: A
authors.initials: A
authors.lastname: Michiels
authors.forename: S
authors.initials: S
authors.lastname: Tuyaerts
authors.forename: A
authors.initials: A
authors.lastname: Bonehill
authors.forename: J
authors.initials: J
authors.lastname: Corthals
authors.forename: K
authors.initials: K
authors.lastname: Breckpot
authors.forename: C
authors.initials: C
authors.lastname: Heirman
authors.forename: S
authors.initials: S
authors.lastname: Van Meirvenne
authors.forename: M
authors.initials: M
authors.lastname: Dullaers
authors.forename: S
authors.initials: S
authors.lastname: Allard
authors.forename: F
authors.initials: F
authors.lastname: Brasseur
authors.forename: P
authors.initials: P
authors.lastname: van der Bruggen
authors.forename: K
authors.initials: K
authors.lastname: Thielemans
mesh_terms: D017951:Antigen Presentation; D019496:Cancer Vaccines / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology; D002470:Cell Survival; D015925:Cryopreservation; D016207:Cytokines / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology*; D018274:Electroporation; D006801:Humans; D016130:Immunophenotyping; D008213:Lymphocyte Activation / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed; D012333:RNA, Messenger / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection
chemical_list: D019496:Cancer Vaccines; D016207:Cytokines; D012333:RNA, Messenger