assayed by the sandwich elisa method human dendritic cells were generated from pbmc cd14 cells were purified by positive selection using anti cd14 microbeads in conjunction with the minimacs system by following the manufacturer 39 s instructions miltenyi biotec auburn ca the cd14 cells were cultured at 1 u00d7 10 6 cells per 1 ml of rpmi 1640 containing 10 fetal bovine serum in 24 well plates with human granulocyte macrophage colony stimulating factor hgm csf 800 u ml and human il 4 500 u ml fresh medium containing hgm csf and il 4 was added every 2 3 days human monocyte derived dendritic cells were used routinely at day 6 of culture cytokines including il 12 in the culture supernatants from dendritic cells and lactobacillus cells were cultured together dendritic cells 1 u00d7 10 6 cells well and lactobacillus cells were incubated at a ratio of 1 10 at 37 u00b0 c for 48 h the culture supernatants were obtained from 48 h cultures the content of il 12 in the culture supernatants was assayed by the sandwich elisa method acid and bile tolerance of the lactic acid bacteria were studied by incubating in mrs broth supplemented with ph 2 5 and 1 5 oxgall the ph was adjusted to 2 5 with hydrochloric acid and lactic acid bacteria were incubated at 37 u00b0 c for 3 h the 1 5 oxgall cultures were incubated at 37 u00b0 c for 4 h each of these five lactobacilli were subcultured at least three times before experimental use followed by centrifugation after the final subculture inoculation into the broth and.