mRNA-electroporated mature dendritic cells retain transgene expression, phenotypical properties and stimulatory capacity after cryopreservation.
Genetically modified dendritic cells (DC) are increasingly used in vitro to activate cytotoxic T lymphocyte (CTL) immune responses. Because T cell activation protocols consist of multiple restimulation cycles of peripheral blood lymphocytes with antigen-loaded mature DC, continuous generation of DC is needed throughout the experiment. Therefore, cryopreservation of DC loaded with antigen is a valuable alternative for weekly generation and modification of DC. Recently, we described an antigen loading method for DC based on electroporation of defined tumor antigen mRNA. In this study, we demonstrate that mRNA-electroporated DC can efficiently be prepared for cryopreservation. Using an optimized maturation and freezing protocol after mRNA electroporation, we obtained high transgene-expressing viable mature DC. In addition, we showed that these modified cryopreserved DC retain stimulatory capacity in an influenza model system. Therefore, cryopreservation of mature mRNA-electroporated DC is a useful method for continuous availability of antigen-loaded DC throughout T cell activation experiments.
Ponsaerts
P
P
Laboratory of Experimental Hematology, Antwerp University Hospital (UZA), University of Antwerp (UIA), Edegem, Belgium.
Van Tendeloo
V F I
VF
Cools
N
N
Van Driessche
A
A
Lardon
F
F
Nijs
G
G
Lenjou
M
M
Mertens
G
G
Van Broeckhoven
C
C
Van Bockstaele
D R
DR
Berneman
Z N
ZN
D017951:Antigen Presentation; D015925:Cryopreservation; D003713:Dendritic Cells / Q000276:immunology; D018274:Electroporation; D018014:Gene Transfer Techniques; D006801:Humans; D007167:Immunotherapy; D020014:K562 Cells; D008213:Lymphocyte Activation; D012333:RNA, Messenger / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology
D012333:RNA, Messenger