Journal: PLoS Neglected Tropical Diseases

Article Title: NLRP3 Controls Trypanosoma cruzi Infection through a Caspase-1-Dependent IL-1R-Independent NO Production

doi: 10.1371/journal.pntd.0002469

Figure Lengend Snippet: PMs from WT mice were infected with T. cruzi Y (3 parasites per cell) (1,5×10 6 cells/300 µl) in the presence of neutralizing antibodies against IL-1β, IL-18 or both cytokines in doses of 0,1, 1 and 10 µg/mL, or left untreated. After 48 h, nitrite accumulation was measured in the culture supernatants by the Griess reaction. Bars represent the mean ± S.D. of triplicate samples where ** p>0,01 and * p>0,05 when compared to untreated group ( A ). T. cruzi -infected PMs from WT and MyD88 −/− mice (3 parasites/cell) were treated or not with z-YVAD-fmk (10 µM). After 48 h, nitrite accumulation was measured in culture supernatants by Griess Rection. Bars represent the mean ± S.D. of triplicate samples where *** p>0,001 and * p>0,01 when compared to untreated group ( B ). PMs (3×10 5 /well of chamber slides) from MyD88 −/− mice were infected with tripomastigotes from T. cruzi Y (5 parasites/cell) for 2–4 h. After removal of extracellular parasites, z-YVAD-fmk (10 µM) was added to cultures. After 48 h chambers were prepared and analyzed as described above to evaluate the numbers of amastigotes. Results were obtained in 1000 counted macrophages from a representative experiment Experiments were repeated three times with the same profile of results ( C ).

Article Snippet: The cells were treated with a caspase-1 inhibitor (z-YVAD-fmk) (10 µM) (MBL International Corporation), an iNOS inhibitor (1 mM) (Aminoguanidin, AG), a cathepsin B inhibitor (Ca-074-Me) (Sigma Aldrich), a K+ channel inhibitor (Glybenclamide, GLB) (InVivogen), an ROS inhibitor (Apocynin, APO) (25 µM to 100 µM) (Sigma Aldrich) or neutralizing antibodies against IL-1β and IL-18 (0,1, 1 and 10 ng/mL) (BD Pharmingen).

Techniques: Infection