Production of cytokines in human MDDC co-cultured with recombinant <i>Lactobacillus</i>.

<p>Monocytes were isolated from human PBMCs by MACS using the Monocyte Isolation Kit II. Monocytes were derived to immature DCs (MDDCs) by cultivating the cells in the presence of 100 nM GM-CSF and 10 nM IL-4 for 5 days. MDDCs (2×10⁵ cells/well) were co-cultured with UV-killed recombinant <i>Lactobacillus</i> expressing OspA (LpA) or the mutant OspA_D17 (LpA_D17) at MOI 10:1 colony-forming units per DC. 100 ng/ml <i>Escherichia coli</i> O111:B4 lipopolysaccharide (LPS) and <i>L. plantarum</i> (Lp) were used as positive and negative control, respectively. After 48 h of stimulation, supernatants were collected and TNF (A), IL-12 (B), IFNγ (C), IL-6 (D) and IL-10 (E) cytokine production was measured by sandwich ELISA (Quantikine). *<i>p</i><0.05,**<i>p</i><0.001. Results are representative of one of three independent experiments.