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. Gene Therapy (2005) 12, 772-782.

Enhanced antigen presentation and CTL activity by transduction of mature rather than immature dendritic cells with octaarginine-modified liposomes

“…Recently, we have reported the novel vaccine delivery system, R8-Lip, which can for CTL induction [24,26]. In the present study, we also found that transduction of mDCs with R8-Lip was preferrential over transduction of iDCs.…”

Efficient gene transfer in CLL by mRNA electroporation

“…20 Electroporation of mRNA is a commonly used method to load dendritic cells with tumor-associated antigens, so they can be used as tumor vaccines. This technique yields high numbers of viable, transgene expressing dendritic cells, [21][22][23][24] adult stem cells, mesenchymal cells, phytohemagglutinin-stimulated T-cells 25 and CD40-activated normal B-cells, 26,27 but, to our knowledge, has not been used for CLL cells.…”

Distinct Effect of CD40 and TNF-Signaling on the Chemokine/Chemokine Receptor Expression and Function of the Human Monocyte-Derived Dendritic Cells

“…It would be the possible reason that immature DCs and conventional cultivated mature DC vaccines was not satisfactory in clinical use. Therefore, further study of the DC migration ability under different maturation stimulating conditions should be important for the design of effective tumor immunotherapy (36,37).…”