MONOCYTE-DERIVED DENDRITIC CELLS - DEVELOPMENT OF A CELLULAR PROCESSOR FOR CLINICAL-APPLICATIONS

Since dendritic cells (DCs) are the most professional antigen-presenti ng cells, (Schuler et al., 1997), increasing interest in their use in clinical approaches has been observed. (Nestle et al., 1998; Murphy G. et al, 1996). We have developed an ex vivo standardized process for t he generation of dendritic-like cells (MAC-DCs) from human blood circu lating monocytes. Human monocytes can differentiate into very differen t functional cells according to the conditions of culture, media and c ytokines used. In the present study, we demonstrate that both pure mon ocytes and mononuclear cells differentiate into DCs when they are grow n in defined medium AIM-V in the presence of granulocyte-macrophage co lony-stimulating factor (GM-CSF) plus IL13 and in approved biocompatib le non-adherent bags. Quality and functional controls of the immature DCs obtained rely on bacterial sterility, viability, morphology and re covery. The MAC-DCs also present an immature DC phenotype with a low e xpression of CD14 and CD64, and high expression of MHC-I MHC-II and CD 40. They also express B7 costimulatory molecules (CD80, CD86), CD83, a nd CD1a molecules. They induce strong allogenic T-cell proliferation ( mixed lymphocyte reaction as well as proliferation of autologous memor y T lymphocytes when incubated in the presence of recall antigens (tub erculosis, Candida albicans, and tetanus toroid). They also show an in crease in phagocytic uptake of yeast, tumour cells and debris. The glo bal closed system which, under reproducible good medical practice (GMP ) conditions, enables the production of dendritic cells of clinical qu ality, has been optimized (''Vac Cell Processor''). It contains all ba gs, connections, media, reagents, washing solutions, control antibodie s, standard operating procedures, data management, traceability and he lp in the form of dedicated software.