In vitro preparation and functional assessment of human monocyte-derived dendritic cells-potential antigen-specific modulators of in vivo immune responses

In this review we present some of our approaches and experiences regarding the isolation of human monocytes from peripheral blood and the in vitro preparation of, first, immature and then mature DCs by applying several maturation factors: bacterial lipopolysaccharide (LPS), a defined mixture of recombinant pro-inflammatory cytokines, monocyte conditioned medium (MCM) and TNF-α alone. The assessment of DC phenotypes and their functional capabilities as well as some of the techniques used for tumour associated antigen loading are also presented. The results of such studies represent a basis for optimal in vitro preparation of DCs, which could be clinically used to modulate immune responses in cancer, autoimmune diseases and in the planned onset of tolerance to disparate major histocompatibilty complex (MHC) antigens prior to tissue or organ transplantation.