After 1 day of culture, IFN-γ production was consistently induced by all strains, except for strains B1697 and B223, and the IFN-γ induction was significantly higher CB-839 price on day 4 compared with that on day 1 (on average 16-fold). A clear difference in IFN-γ induction was observed for the different strains tested, with strains B1697 and B223 eliciting consistently low IFN-γ induction whereas the other strains were strong inducers. The strong

IFN-γ-inducing strains also showed an increased IL-12 production, though IL-12 levels were, in all samples, below 25 pg mL−1 (data not shown). IL-13 could not be detected on day 1 and was <25 pg mL−1 on day 4. To determine the effects of lactobacilli interacting with stimulated hPBMC, αCD3/αCD28 was added to the culture and cells were cultured for 4 days. All strains inhibited IL-13 production by αCD3/αCD28-stimulated hPBMC (Fig. 4f). Strain B2261, the mixture

of strains B2261 and B633, and strain B633 alone were significantly stronger IL-13 inhibitors (on average a factor 7 inhibition) compared with the other strains tested (on average a factor 3 inhibition). There was a clear tendency of lactobacilli to inhibit IL-1β production, except for strains B1697 and B223 (Fig. 4a), while TNF-α (Fig. 4c) and IL-10 (Fig. 4b) production was increased compared with the control for most strains, except for strains B223 and CBI 118. Addition of the various Lactobacillus strains to the hPBMC had no effect on IFN-γ production, which was high in all cultures after stimulation CP-690550 concentration Methane monooxygenase with αCD3/αCD28 (Fig. 4d). IL-12 (Fig. 4e) was induced by strains B1836, B2261, the mixture of B2261 and B633, B633 alone and CBI 118, which was the same group of strains that also induced

IL-12 and IFN-γ production in the unstimulated cultures. The polyclonal stimulus αCD3/αCD28 clearly induced IL-1β, IL-10, TNF-α, IFN-γ and IL-13 production compared with the unstimulated cultures. The induction of IL-10 by the strains was significantly lower in the αCD3/αCD28-stimulated cultures compared with the unstimulated cultures for the mixture of strains B2261 and B633, and strain B633. To determine the effect of the different lactobacilli on antigen-specific stimulated cultures, hPBMC of the five birch pollen-allergic patients were cultured in the presence of the major birch pollen allergen Bet v 1 and in the presence or absence of the different lactobacilli. After 8 days of culture, four stimulation conditions were compared. The restimulation condition with αCD3/αCD28 on day 7 was used to increase the amount of antigen-specific T cells in the cultures, which are still expected to be active in the culture and proliferate upon interaction with the specific antigen, Bet v 1. The addition of Bet v 1 did not result in significant differences in cytokine production profiles compared with the medium control.