jensenii selleck products derivatives (Figure 4). Again, MALP-2, in contrast to L. jensenii, induced a significant IL-8 upregulation in all three

models. Since the findings in the primary tissue model (Figure 4a) mirrored those in the immortalized epithelial monolayers (Figure 3b and 4b), as previously reported with other vaginal bacteria [20], we chose the immortalized cell line model for further analysis of immunity mediators and CFU counts based on its lower cost- and handling time efficiency. Figure 4 Cytokine profiles induced by bacteria or synthetic TLR2/6 ligand in cervicovaginal colonized epithelial model. KPT-330 in vitro Similar IL-8 levels measured in supernatants derived from primary and immortalized epithelial cells cultured with L. jensenii

1153–1666, 3666, gfp bioengineered and L. jensenii 1153 wild type (WT) strains or MALP-2 50 nM as a positive control. (Figure 4a) Two independent experiments with (VEC-100™) primary ectocervical originated tissue. (Figure 4b) Vaginal (Vk2/E6E7) and endocervical (End1/E6E7) epithelial colonized cells in one representative of three experiments. Bars represent mean and SEM from duplicate cultures. *** P<0.001 different from medium control, +++ P<0.001 different from L. jensenii WT. In further immune mediator analysis of L. jensenii colonized Vk2/E6E7 immortalized epithelial monolayers; MALP-2 induced significant increases over baseline levels of TNF-α (P<0.001) and IL-6 LXH254 datasheet (P<0.001), while the WT and derivatives had no significant effect on either (Figure

5a-b). IL-1α levels slightly increased (P<0.05) in the presence of the WT, however all derivatives maintained baseline levels (Figure 5c). No significant differences were observed in IL-1RA levels (Figure 5D). Figure selleck kinase inhibitor 5 Absence of a pro-inflammatory cytokine response in L. jensenii colonized epithelial model. (Figure 5a) TNF-α, (Figure 5b) IL-6, (Figure 5c) IL-1α, (Figure 5d) IL-1RA cytokine levels measured in supernatants from vaginal (Vk2/E6E7) epithelium cultured for 24 h with L. jensenii 1153–1666, 3666, and gfp bioengineered strains and L. jensenii 1153 wild (WT) strain or MALP-2 (50 nM) as a positive control. Bars represent mean and SEM from duplicate and triplicate cultures in two independent experiments. *** P<0.001,* P<0.05 different from medium control, +++ P<0.001 different from L. jensenii 1153 WT. Sustained bacterial colonization by wild type and bioengineered L. jensenii does not alter levels of inflammation-associated proteins over time To determine if the homeostatic effect of L. jensenii on innate immunity proteins is sustained over time, despite NF-κB activation, we exposed the vaginal epithelial cells to wild type and bioengineered bacterial strains and MALP-2 and maintained the cultures for three days with supernatants harvested for protein measurement and replaced with plain KSFM medium at each 24 h interval.