2b). The ‘patchiness’ of the lesions complicated the scoring and, as a consequence, no

significant differences in histological scoring could be observed between the treatment groups (Table 1). Similarly, cytokine analysis by qRT-PCR did not reveal significant differences (data not shown). As described above, in the 3% DSS-induced model the epithelial layer was severely damaged with patchy lesions (Fig. 2b), and the administration of LGG wild-type was shown to be detrimental, in contrast to administration of the dltD GSK-3 cancer mutant (Fig. 2a and Table 1). Because Yan et al. [25] reported that the intestinal epithelial cells are an important target for certain probiotic actions of LGG, we investigated subsequently whether the detrimental effect of LGG wild-type and the enhanced efficacy of the dltD mutant correlated with the integrity of the intestinal barrier. Hereto, C57/BL6 mice received 1% DSS for three cycles of 7 days DSS–7 days normal drinking water. LGG

wild-type and dltD mutant were then given in the drinking water starting 3 Dabrafenib price days before colitis induction. In this model, there was no significant difference in body weight between the PBS-, wild-type- and dltD-treated groups (Fig. 2c). However, the dltD-mutant treated group showed a significantly attenuated colonic inflammation based on the macroscopic score (Table 2). In this milder model, epithelial damage was much less pronounced than in the 3% DSS-induced model, although colitis lesions were still clearly visible (Fig. 2d). Interestingly, the LGG wild-type also showed a trend of ameliorating the severity of the colitic parameters in this mild chronic model, although no significant difference could be observed compared to the PBS-treated group. qRT-PCR results revealed that the administration of the dltD mutant reduced mucosal IL-12p40 mRNA expression compared to the PBS-treated (P = 0·0170) and LGG wild-type-treated groups (P = 0·0363) (Fig. 3a).

IFN-γ expression was also reduced in the dltD-treated group and this was significant compared to the PBS-treated group (P = 0·0276) (Fig. 3b). As these differences in cytokine expression might be downstream GNA12 effects of a different TLR expression, we subsequently determined TLR-1, TLR-2, TLR-4 and TLR-6 mRNA expression in the three treatment groups. Mice treated with the dltD mutant showed a reduced expression of TLR-2 compared to PBS-treated mice (P = 0·0006) (Fig. 3c). Compared to LGG wild-type-treated mice, we also observed lower expression levels of TLR-1 (P = 0·0179), TLR-2 (P = 0·0328) and TLR-4 (P = 0·0443) in the dltD treated group (Fig. 3c–e). No significant differences in cytokines TNF, IL-1β, IL-10 and TGF-β were seen (data not shown). Also no significant changes in expression of TLR-6 (Fig. 3f) were observed between the three treatment groups.