Journal: Rheumatology (Oxford, England)

Article Title: Expression of sterile-α and armadillo motif containing protein (SARM) in rheumatoid arthritis monocytes correlates with TLR2-induced IL-1β and disease activity

doi: 10.1093/rheumatology/keab162

Figure Lengend Snippet: TLR1/2 activation induces higher IL-1β secretion from RA patient monocytes than healthy controls CD14 + monocytes were isolated from peripheral blood donated by HC and RA patients. IL-1β secretion was measured following 24 hours of culture with media alone (unstimulated) and ( A ) 100 ng/ml Pam 3 CSK 4 (HC n = 18; RA n = 28), ( B ) 10 ng/ml LPS (HC n = 21; RA n = 29) or ( C ) 2 μg/ml R-848 (HC n = 22; RA n = 28). Data is shown as the mean ± SEM. Significance was tested using two-tailed Mann–Whitney U test (*** P = 0.0009). HC: healthy control; LPS: lipopolysaccharide; TLR: Toll-like receptor.

Article Snippet: Peripheral blood monocytes from PBMCs derived from leucocyte cones were isolated by iso-osmotic Percoll gradient centrifugation, as previously described [ ], before being cultured in RPMI1640 media supplemented with 5% (v/v) foetal calf serum and 1% (v/v) penicillin/streptomycin solution (PAA, Pasching, Austria) with or without 100 ng/ml PAM 3 CSK 4 (Axxora, Nottingham, UK), 10 ng/ml lipopolysaccharide (LPS) (Axxora, Nottingham, UK) or 2 μg/ml resiquimod (R-848) (Enzo Life Sciences, Lausen, Switzerland) at 37°C, 5% CO 2 .

Techniques: Activation Assay, Isolation, Two Tailed Test, MANN-WHITNEY

Journal: Rheumatology (Oxford, England)

Article Title: Expression of sterile-α and armadillo motif containing protein (SARM) in rheumatoid arthritis monocytes correlates with TLR2-induced IL-1β and disease activity

doi: 10.1093/rheumatology/keab162

Figure Lengend Snippet: SARM expression negatively correlates with IL-1β secretion following TLR1/2 stimulation in RA monocytes TLR1/2-induced IL-1β secretion was measured following 24 hours of stimulation with 100 ng/ml Pam 3 CSK 4 and correlated with the basal expression of ( A ) NLRP3 ( n = 22), ( B ) CASP1 ( n = 22), ( C ) PYCARD ( n = 22) and ( D ) SARM1 ( n = 22) in monocytes from RA patients. The significance was analysed using a two-tailed Spearman test. ( E ) Monocyte IL1B expression following a 3-hour stimulation with 100 ng/ml Pam 3 CSK 4 was compared with basal SARM1 expression (prior to stimulation) in matched patient samples ( n = 12). ( F ) Monocytes from healthy volunteers ( n = 7) were stimulated for 6 hours with 100 ng/ml Pam 3 CSK 4 , 10 ng/ml LPS or 2 µg/ml R-848. SARM1 expression was expressed as the fold change from US cells and shown as the mean ± SEM. A one-way ANOVA using Dunnett’s multiple comparisons test was used to test significance compared with US cells (** P = 0.007, **** P < 0.0001). LPS: lipopolysaccharide; SARM: sterile-α and armadillo motif containing protein; TLR: Toll-like receptor; US: unstimulated.

Article Snippet: Peripheral blood monocytes from PBMCs derived from leucocyte cones were isolated by iso-osmotic Percoll gradient centrifugation, as previously described [ ], before being cultured in RPMI1640 media supplemented with 5% (v/v) foetal calf serum and 1% (v/v) penicillin/streptomycin solution (PAA, Pasching, Austria) with or without 100 ng/ml PAM 3 CSK 4 (Axxora, Nottingham, UK), 10 ng/ml lipopolysaccharide (LPS) (Axxora, Nottingham, UK) or 2 μg/ml resiquimod (R-848) (Enzo Life Sciences, Lausen, Switzerland) at 37°C, 5% CO 2 .

Techniques: Expressing, Two Tailed Test