Figure 6. IL-6 production by explanted splenocytes and BMDM from Ity, Ity3, Ity3.RecN and Ity3.RecG after stimulation with PAMPs or infection with Salmonella Typhimurium.

A) IL-6 production by explanted splenocytes after stimulation with flagellin (Flg), lipoteichoic acid (LTA), lipopolysaccharide (LPS), zymosan (Zym), CpG-DNA (CpG), and PolyIC. The culture medium (Med) was used as negative control. Ity (n = 4), Ity3 (n = 4), Ity3.RecN (n = 4) and Ity3.RecG (n = 4). Significant differences in IL-6 production are shown with respect to the reference Ity strain. Splenocytes from Ity3.RecG strain were hypo-responsive to stimulation with Tlr4 and Tlr5 ligands as shown by significant lower levels of IL-6 after stimulation with LPS and flagellin, respectively, as compared to the Ity strain. No major differences were seen in IL-6 production in Ity3 and Ity3.RecN strains implying that these strains do not have a defect in TLR-dependent activation of IL-6 production. The data shown are representative of two experiments with n = 4 per experiment B) IL-6 production measured in the supernatant of BMDM derived from Ity (n = 4), Ity3 (n = 4), Ity3.RecN (n = 4) and Ity3.RecG (n = 4) male mice. The data shown are representative of three experiments with n = 4 per experiment. BMDM were infected with Salmonella Typhimurium. After a period of 45 min, RPMI 1640 containing 100 µg/ml gentamycin was added for a period of 1 h (T0). The medium was subsequently replaced by RPMI 1640 containing 10 µg/ml gentamycin and IL-6 was measured after 2 (T2), 4 (T4) and 6 (T6) hrs of incubation. Significant differences in IL-6 production are shown with respect to the reference Ity strain. The data were analysed by two-way ANOVA followed by multiple-comparisons testing. *corresponds to a P-value of ≤0.05, **corresponds to a P-value of ≤0.01, ***corresponds to P-value ≤0.001, while ****implies a P≤0.0001.