Hp infection elicits IL-1β secretion.

<p>(<b>A–E</b>) Mice were administered 200 L3 Hp by oral gavage. IL-1β protein levels were measured by ELISA in (<b>A</b>) the peritoneal wash and (<b>B</b>) intestinal tissue culture supernatants at the indicated timepoints. (<b>C</b>) Hp worm counts were performed for defined segments of the small intestine from WT (C57BL/6) at 13 dpi. (<b>D</b>) IL-1β was measured by ELISA for duodenum tissue culture supernatants of day 6 infected bone marrow chimera mice (donor strain/recipient strain). Results are representative of at least 3 independent experiments (n = 5 per group) and expressed as mean ± SEM. (<b>E</b>) Intestinal lamina propria CD11b⁺ and CD11b⁻ cells were isolated at 6 dpi and analyzed by Western blot for IL-1β expression. Band intensity for IL-1β and β-actin was determined using Adobe Photoshop CS3. The ratio between IL-1β and the control protein β-actin band intensity was then determined. IL-1β is expressed as percent of β-actin intensity. (<b>F</b>) BMMs from WT (C57BL/6), Trif<i>^−/−</i> or MyD88<i>^−/−</i> mice were stimulated <i>in vitro</i> with HPL5 (100 µg/mL), HES (5 µg/mL) or P.HES (5 µg/mL) for 4 hours and cell extracts were analyzed for pro-IL-1β expression by Western blot. (<b>G</b>) BMMs from WT (C57BL/6), Asc<i>^−/−</i> or Nlrp<i>3^−/−</i> mice were stimulated <i>in vitro</i> with P.HES, HPL5 or LPS plus MSU and culture supernatants (SN) were analyzed for active IL-1β by ELISA. Cell extracts (XT) and SN were also analyzed for pro- or mature- IL-1β by Western blot. Pro-Casp1 was used as a control protein. XT blots for IL-1β and Casp1 demonstrate the pro-forms, and the observed lower bands in the IL-1β and Casp1 blots are cleavage products. Error bars represent means of triplicate cultures ± SEM and the experiment was repeated 3 times.