Three-day dendritic cells for vaccine development: Antigen uptake, processing and presentation | Burdek et al. Journal of Translational Medicine 2010 8 90 http content 8 1 90 JOURNAL OF TRANSLATIONAL MEDICINE RESEARCH Open Access Three-day dendritic cells for vaccine development Antigen uptake processing and presentation Maja Burdek Stefani Spranger Susanne Wilde Bernhard Frankenberger Dolores J Schendel Christiane Geiger Abstract Background Antigen-loaded dendritic cells DC are capable of priming naive T cells and therefore represent an attractive adjuvant for vaccine development in anti-tumor immunotherapy. Numerous protocols have been described to date using different maturation cocktails and time periods for the induction of mature DC mDC in vitro. For clinical application the use of mDC that can be generated in only three days saves on the costs of cytokines needed for large scale vaccine cell production and provides a method to produce cells within a standard work-week schedule in a GMP facility. Methods In this study we addressed the properties of antigen uptake processing and presentation by monocyte-derived DC prepared in three days 3d mDC compared with conventional DC prepared in seven days 7d mDC which represent the most common form of DC used for vaccines to date. Results Although they showed a reduced capacity for spontaneous antigen uptake 3d mDC displayed higher capacity for stimulation of T cells after loading with an extended synthetic peptide that requires processing for MHC binding indicating they were more efficient at antigen processing than 7d DC. We found however that 3d DC were less efficient at expressing protein after introduction of in vitro transcribed ivt RNA by electroporation based on published procedures. This deficit was overcome by altering electroporation parameters which led to improved protein expression and capacity for T cell stimulation using low amounts of ivtRNA. Conclusions This new procedure allows 3d mDC to replace 7d mDC for use in DC-based vaccines that utilize long peptides proteins
