Journal: Blood
Article Title: NLRP3 inflammasome blockade treats intestinal inflammation associated with chronic granulomatous disease
doi: 10.1182/blood.2025029676
Figure Lengend Snippet: βHB inhibits NLRP3 inflammasome activity in human cells in the context of CGD. Western blot analysis of IL-1β (pro–IL-1β and active IL-1β) and β-actin (loading control) expression in WT (left) compared to CYBB −/− (ie, CGD; right) THP-1 cells (A) or PBMCs from a healthy person and a patient with CGD (C) that were either unstimulated, treated with the nigericin trigger (1 hour) alone (nigericin), or stimulated with lipopolysaccharide (LPS; 3 hours) and nigericin (1 hour) with or without NLRP3 inhibition with different doses of βHB (1mM-20mM) 30 minutes before adding the nigericin trigger. The last lane shows cells treated with 10mM βHB alone (ie, no priming and no trigger). The measurement of IL-1β, IL-6, and TNF levels in cell culture supernatants after the treatment of WT and CYBB −/− THP-1 cells (B) or healthy and CGD PBMCs (D) with either PBS (unstimulated), LPS (3 hours) and nigericin (1 hour; stimulated), or LPS followed by NLRP3 inhibition with 20mM βHB before (stimulated + βHB) or 10μM MCC950 (stimulated + MCC950) for 30 minutes before adding nigericin. Data in panels C-D are representative of 2 independent experiments. Significance was determined using multiple Mann-Whitney U tests for panel C or unpaired t tests for panel D, with data represented as median ± IQR or mean ± SEM, respectively. Stim., stimulated; Unstim., unstimulated.
Article Snippet: In vitro experiments were performed using human monocytic leukemia THP-1 cell lines, including WT THP-1 (TIB-202; American Type Culture Collection) and NOX2-deficient ( CYBB −/− ) THP-1 cells (generously provided by Fabien Touzot, CHU Sainte-Justine ), cultured in RPMI 1640 (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (Gibco) and 1% penicillin-streptomycin-glutamine (Gibco).
Techniques: Activity Assay, Western Blot, Control, Expressing, Inhibition, Cell Culture, MANN-WHITNEY
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