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

<p>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. <i>Ity</i> (n = 4), <i>Ity3</i> (n = 4), <i>Ity3.RecN</i> (n = 4) and <i>Ity3.RecG</i> (n = 4). Significant differences in IL-6 production are shown with respect to the reference <i>Ity</i> strain. Splenocytes from <i>Ity3.RecG</i> strain were hypo-responsive to stimulation with <i>Tlr4</i> and <i>Tlr5</i> ligands as shown by significant lower levels of IL-6 after stimulation with LPS and flagellin, respectively, as compared to the <i>Ity</i> strain. No major differences were seen in IL-6 production in <i>Ity3</i> and <i>Ity3.RecN</i> 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 <i>Ity</i> (n = 4), <i>Ity3</i> (n = 4), <i>Ity3.RecN</i> (n = 4) and <i>Ity3.RecG</i> (n = 4) male mice. The data shown are representative of three experiments with n = 4 per experiment. BMDM were infected with <i>Salmonella</i> 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 <i>Ity</i> 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.