%0 Journal Article
%T A full scale comparative study of methods for generation of functional Dendritic cells for use as cancer vaccines
%A Silvija Jarnjak-Jankovic
%A Hege Hammerstad
%A Stein S£¿b£¿e-Larssen
%A Gunnar Kvalheim
%A Gustav Gaudernack
%J BMC Cancer
%D 2007
%I BioMed Central
%R 10.1186/1471-2407-7-119
%X The Elutra Cell Selection System was used to isolate monocytes after collection of leukapheresis product. The enriched monocytes were cultured in gas permeable Teflon bags with IL-4 and GM-CSF for 24 hours (Fast DC) or 5 days (Standard DC) to obtain immature DCs. The cells were then transfected with mRNA from the leukemia cell line Jurkat E6 by electroporation and incubated for additional 24 h or 2 days in the presence of pro-inflammatory cytokines (TNF¦Á, IL-1¦Â, IL-6 and PGE2) to obtain mature DCs.Mature Fast DC and Standard DC displayed comparable levels of many markers expressed on DC, including HLA-DR, CD83, CD86, CD208 and CCR7. However, compared to Standard DC, mature Fast DC was CD14high CD209low. Fast DC and Standard DC transfected with Jurkat E6-cell mRNA were equally able to elicit T cell specifically recognizing transfected DCs in vitro. IFN¦Ã-secreting T cells were observed in both the CD4+ and CD8+ subsets.Our results indicate that mature Fast DC are functional antigen presenting cells (APCs) capable of inducing primary T-cell responses, and suggest that these cells may be valuable for generation of anti-tumor vaccines.Dendritic cells (DCs) are potent antigen-presenting cells (APCs) involved in the induction of T-cell-mediated immune responses and have appeared as important candidates for cellular-based therapies [1]. Immature DCs are found throughout the body where they capture and process antigens (Ag). When activated by danger signals the DCs start differentiation towards a mature penotype and migration to the T cell dependent areas of secondary lymphoid organs. During this process, they lose the capacity for Ag-capturing and upregulate major histocompatibility complex (MHC)- and costimulatory molecules for stimulation of naive T cells [2-4].In vitro, DCs can be differentiated from various cellular sources, including CD34+ progenitor cells from bone marrow (BM) and cord blood (CB), and monocytes obtained from peripheral blood mononuclear cells (PBMCs)
%U http://www.biomedcentral.com/1471-2407/7/119