GENERATION OF DENDRITIC CELLS FROM PERIPHERAL-BLOOD ADHERENT CELLS INMEDIUM WITH HUMAN SERUM

Dendritic cells (DC) provide an effective pathway for presenting antig ens to T cells, both self-antigens during T-cell development and forei gn antigens during immunity. As such, these cells may be promising adj uvants for immunotherapy. Thus, it is important to establish simple an d fast method(s) to generate sufficient numbers of human DC in medium free of calf serum so that the cells can be used for both experimental and clinical purposes. In this report, we used peripheral blood adher ent cells, without laborious cell purification or depletion, as the st arting population and cultured them in medium supplemented with granul ocyte/macrophage colony-stimulating factor and interleukin-4. Substant ial numbers of cells with the phenotypical and functional characterist ics of immature DC were obtained in a 7-day culture. We then compared DC cultured in medium supplemented with either fetal calf serum or poo led human ABRh(+) serum and found no difference in cell yields and in their ability to stimulate alloreactive T cells or to present soluble antigens to T cells. Irradiated cells were less efficient than non-irr adiated cells in antigen presentation and stimulation of T cells. Fina lly, we have examined DC with or without additional tumour necrosis fa ctor-alpha treatment and found that antigen-pulsed mature cells could as efficiently present antigen to T cells as did immature cells. This method is suitable for the generation of DC in studies of large clinic al materials.