Journal: Scientific Reports

Article Title: GWAS-identified hyperuricemia-associated IGF1R variant rs6598541 has a limited role in urate mediated inflammation in human mononuclear cells

doi: 10.1038/s41598-024-53209-7

Figure Lengend Snippet: Role of IGF1R in urate induced inflammation in vitro. ( A ) Freshly isolated PBMCs isolated from healthy controls (n = 9) were stimulated with RPMI, uric acid 50 mg/dl, IGF-1 recombinant protein (R&D Systems) 5 ng for 24 h. After 24 h the cells were restimulated with LPS 10 ng/ml together with MSU 300 mg/dl. ( B ) Freshly isolated Percoll enriched Monocytes (n = 6) were treated with RPMI, IGF1R antibody (R&D Systems) or IgG1 isotype control and uric acid 50 mg/dl for 24 h. After 24 h monocytes were restimulated with LPS 10 ng/ml with MSU 300 mg/dl. ( C ) Freshly isolated PBMCs (n = 6) were trained in vitro with IGF-1 5 ng and beta-glucan (BG) 1 μg/ml for 24 h, subsequently washed, rested for 5 days, and at day 6 restimulated for 24 h with 10 ng/ml LPS. ( D ) Freshly isolated PBMCs originating from healthy controls (n = 174). Concentration IL-1β and IL-6 measured in the supernatants of PBMCs after stimulation with urate of conc. 50 mg/dl and 12.5 mg/dl for 24 h, followed by restimulation with LPS 10 ng/ml. IL-1β, and IL-1Ra (R&D Systems, Minneapolis) was measured in supernatant by ELISA. Graphs depict means ± SEM. Friedman test, Dunn's multiple comparisons test, p* < 0.05.

Article Snippet: Cytokine concentrations were determined in cell culture supernatants using specific sandwich ELISA kits for IL-1β, IL-1Ra, IL-6 (R&D Systems, Minneapolis).

Techniques: In Vitro, Isolation, Recombinant, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay