Journal: In Vivo

Article Title: Dysregulated Pro-inflammatory and Anti-inflammatory Cytokine Responses to Microbe-associated Molecular Patterns in X-linked Chronic Granulomatous Disease

doi: 10.21873/invivo.13989

Figure Lengend Snippet: Production of cytokines and chemokines associated with innate immunity by peripheral blood mononuclear cells (PBMCs) isolated from healthy controls and patients with X-linked chronic granulomatous disease (CGD). PBMCs (1×10 6 cells/ml) isolated from healthy controls (n=14 or 12) and patients with X-linked CGD (n=2) were stimulated with FK156 (20 μg/ml), MDP (20 μg/ml), PAM (1 μg/ml), LPS (1 μg/ml), zymosan (5 μg/ml), curdlan (50 μg/ml), HKCA (1×10 8 cells/ml), HKSC (1×10 8 cells/ml), or MSU (100 μg/ml) for 72 h. Culture supernatants were subjected to enzyme-linked immunosorbent assay to measure (A) CXCL8, (B) IL-1β, (C) IL-6, and (D) TNF-α concentrations. Results are presented as the mean±standard error of mean. Each dot presents a value obtained from each subject. *p<0.05, **p<0.01, as compared with healthy control-derived PBMCs stimulated with each ligand. MDP: Muramyl dipeptide; PAM: PAM3CSK4; LPS: lipopolysaccharide; HKCA: heat-killed Candida albicans; HKSC: heat-killed Saccharomyces cerevisiae; MSU: monosodium urate crystals; CXCL8: C-X-C motif chemokine ligand 8; IL-1β: interleukin-1β; TNF-α: tumor necrosis factor-α.

Article Snippet: Cells were stimulated with FK156 (20 μg/ml; Astellas Pharma, Tokyo, Japan), muramyl dipeptide (MDP, 20 μg/ml; InvivoGen, San Diego, CA, USA), PAM 3 CSK4 (PAM, 1 μg/ml; InvivoGen), lipopolysaccharide (LPS, 1 μg/ml; Sigma-Aldrich, St. Louis, MO, USA), zymosan (5 μg/ml, InvivoGen), curdlan (50 μg/ml, Sigma-Aldrich), heat-killed Candida albicans (HKCA, 1×10 8 cells/ml; InvivoGen), heat-killed Saccharomyces cerevisiae (HKSC, 1×10 8 cells/ml; InvivoGen), and monosodium urate crystals (MSU, 100 μg/ml; InvivoGen) as described previously ( , , , ).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Control, Derivative Assay

Journal: In Vivo

Article Title: Dysregulated Pro-inflammatory and Anti-inflammatory Cytokine Responses to Microbe-associated Molecular Patterns in X-linked Chronic Granulomatous Disease

doi: 10.21873/invivo.13989

Figure Lengend Snippet: Production of cytokines, IgG1, and IgG4 by peripheral blood mononuclear cells (PBMCs) isolated from healthy controls and patients with X-linked chronic granulomatous disease (CGD). PBMCs (1×10 6 cells/ml) isolated from healthy controls (n=18, 16, 14, or 12) and patients with X-linked CGD (n=2) were stimulated with FK156 (20 μg/ml), MDP (20 μg/ml), PAM (1 μg/ml), LPS (1 μg/ml), zymosan (5 μg/ml), curdlan (50 μg/ml), HKCA, 1×10 8 cells/ml), HKSC (1×10 8 cells/ml), or MSU (100 μg/ml) for 72 h or seven days. Culture supernatants were subjected to enzyme-linked immunosorbent assay to measure (A) IFN-γ, (B) IL-10, (C) IgG1, and (D) IgG4 concentrations. Results are presented as the mean±standard error of mean. Each dot presents a value obtained from each subject. *p<0.05, **p<0.01, compared with healthy control-derived PBMCs stimulated with each ligand. MDP: Muramyl dipeptide; PAM: PAM3CSK4; LPS: lipopolysaccharide; HKCA: heat-killed Candida albicans; HKSC: heat-killed Saccharomyces cerevisiae; MSU: monosodium urate crystals; IFN-γ: interferon-γ; IL-10: interleukin-10; IgG1: immunoglobulin G1; IgG4: immunoglobulin G4.

Article Snippet: Cells were stimulated with FK156 (20 μg/ml; Astellas Pharma, Tokyo, Japan), muramyl dipeptide (MDP, 20 μg/ml; InvivoGen, San Diego, CA, USA), PAM 3 CSK4 (PAM, 1 μg/ml; InvivoGen), lipopolysaccharide (LPS, 1 μg/ml; Sigma-Aldrich, St. Louis, MO, USA), zymosan (5 μg/ml, InvivoGen), curdlan (50 μg/ml, Sigma-Aldrich), heat-killed Candida albicans (HKCA, 1×10 8 cells/ml; InvivoGen), heat-killed Saccharomyces cerevisiae (HKSC, 1×10 8 cells/ml; InvivoGen), and monosodium urate crystals (MSU, 100 μg/ml; InvivoGen) as described previously ( , , , ).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Control, Derivative Assay