RNA-containing adenovirus/polyethylenimine transfer complexes effectively transduce dendritic cells and induce antigen-specific T cell responses.
BACKGROUND
Dendritic cells (DCs) are the most potent antigen-presenting cells in initiating primary immune responses. Given the unique properties of DCs, gene-modified DCs represent a particularly attractive approach for immunotherapy of diseases such as cancer.


METHODS
Gene-modified DCs were obtained by a receptor-mediated gene delivery system using adenovirus (Ad) particles as ligand and RNA or DNA condensed by polyethylenimine (PEI). In vitro transcribed polyadenylated or non-polyadenylated RNA was used. RNA-transduced DCs were generated expressing chicken ovalbumin (OVA) or chimeric constructs thereof, and compared with DNA-transduced DCs.


RESULTS
Ad/PEI transfection complexes efficiently delivered RNA into DCs. Such RNA-transduced DCs induced OVA-specific T cell responses more effectively than DNA-transduced DCs. Furthermore, DCs transduced with polyadenylated RNA were more potent in stimulating CD4(+) and CD8(+) T cell responses than DCs transduced with non-polyadenylated RNA and this was particularly important for CD4(+) T cell responses.


CONCLUSIONS
Ad/PEI/RNA transfection is an efficient means for generating RNA-transduced DCs and for stimulating antigen-specific T cell responses. Polyadenylation of RNA enhances CD8(+) T cell responses and is essential for CD4(+) T cell responses.
Gust
Tatjana C
TC
Max-Delbrück-Center for Molecular Medicine, MDC, Robert-Rössle-Str. 10, D-13092 Berlin, Germany.
Diebold
Sandra S
SS
Cotten
Matt
M
Zenke
Martin
M
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D000941:Antigens / Q000235:genetics; D015496:CD4-Positive T-Lymphocytes / Q000502:physiology; D018414:CD8-Positive T-Lymphocytes / Q000502:physiology; D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage; D003713:Dendritic Cells / Q000502:physiology*; D000949:Histocompatibility Antigens Class II / Q000235:genetics / Q000276:immunology; D008213:Lymphocyte Activation / Q000235:genetics; D046911:Macromolecular Substances; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000235:genetics; D026723:Polyadenylation; D011094:Polyethyleneimine; D012313:RNA / Q000008:administration & dosage* / Q000737:chemistry; D013601:T-Lymphocytes / Q000276:immunology* / Q000502:physiology*; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection / Q000379:methods
D000941:Antigens; D000949:Histocompatibility Antigens Class II; D046911:Macromolecular Substances; D012313:RNA; D011094:Polyethyleneimine; D010047:Ovalbumin; D004247:DNA