albicans the suspension was diluted to the appropriate concentration with pyrogen free saline microscopy confirmed that the suspension consisted of blastoconidia when necessary the blastoconidia were heat killed either at 56 u00b0 c for 1 hour or at 100 u00b0 c for 30 minutes immature dendritic cells imdc were generated from human peripheral blood monocytes as described previously vissers et al 2001 briefly monocytes were isolated by adherence to plastic and cultured in the presence of il 4 500 u ml and gm csf 800 u ml for 6 7 days k562 transfectants either expressing dc sign or alcam were generated by transfection of k562 cells with 10 u03bcg of plasmid by electroporation as described previously geijtenbeek et al 2000b nelissen et al 2000 positive cells were sorted several times to obtain stable transfectants with similar expression levels of dc sign labeling of candida cells was performed as follows viable or heat killed 60 minutes at 56 u00b0 c or 30 minutes at 100 u00b0 c yeast cells were resuspended to 2 u00d7 10 8 ml in 0 1 mg ml fitc in 0 05 m carbonate bicarbonate buffer ph 9 5 after incubation for 15 minutes at room temperature in the dark fitc labeled candida cells were washed twice in pbs containing.