Interleukin (IL)-31 activates signal transducer and activator of transcription (STAT)-1, STAT-5 and extracellular signal-regulated kinase 1/2 and down-regulates IL-12p40 production in activated human macrophages.

BACKGROUND: Interleukin-31 is a cytokine expressed by activated T cells. A major function of IL-31 in atopic dermatitis (AD) is the induction of pruritus in the skin. We recently showed that IL-31 induces pro-inflammatory cytokines following staphylococcal exotoxins' stimulation in human macrophages. However, signalling pathways of IL-31 in activated human macrophages still remain unclear. The aim of the study was to investigate the signalling pathways of IL-31 receptor as well as functional effects of IL-31 in activated macrophages. METHODS: Human macrophages were prestimulated with staphylococcal exotoxins (SEB, α-toxin) to up-regulate the IL-31 receptor with and without IL-31. Phospho-signal transducer and activator of transcription (pSTAT) 1/3/5, phospho-extracellular signal-regulated kinase (ERK 1/2), ß-actin as well as p21/WAF/Cip1 levels were determined by means of Western blot analysis. Interleukin-12p40, IL-12p70 and IL-23 secretions were assessed by using an enzyme-linked immunosorbent assay. RESULTS: Interleukin-31 strongly activated STAT-1 and 5 but not STAT-3 in human macrophages after up-regulation of IL-31 receptor with staphylococcal exotoxins. p21/WAF/Cip1 expression was induced by IL-31 in activated human macrophages. Furthermore, IL-31 down-regulated. IL-12p40 secretion via ERK 1/2 phosphorylation in human macrophages following up-regulation of IL-31 receptor with staphylococcal exotoxins. CONCLUSIONS: The T helper (Th) 2 cytokine IL-31 induces pro-inflammatory effects in activated human macrophages via STAT-1 and 5 phosphorylation. Interleukin-31-induced ERK 1/2 activation contributes to the underlying mechanism of Th1 cytokine IL-12 suppression in macrophages. This mechanism may be relevant in Th2 inflammatory responses and may help to develop therapeutic strategies in IL-31-associated diseases such as AD.

Macrophages from patients with atopic dermatitis show a reduced CXCL10 expression in response to staphylococcal α-toxin.

BACKGROUND: Patients with atopic dermatitis (AD) are frequently colonized with Staphylococcus aureus (S. aureus), one-third of them producing α-toxin, which is correlated with the severity of eczema in AD. Staphylococcus aureus colonizes in patients with psoriasis as well. Distinct expression of chemokine (C-C motif) ligand (CCL) and chemokine (C-X-C motif) ligand (CXCL) chemokines has been documented in both diseases. In this study, we investigated the effects of sublytic α-toxin concentrations on human macrophages that accumulate in the skin of patients with AD and psoriasis. METHODS: IFN-γ-induced protein of 10-kDa (IP-10)/CXCL10 and macrophage-derived chemokine (MDC)/CCL22 production were evaluated at the mRNA or at the protein level using qRT-PCR or ELISA, respectively. Cell surface markers' expression and chemotaxis were determined by flow cytometry and Boyden chamber technique, respectively. RESULTS: Sublytic concentrations of α-toxin strongly induced CXCL10 in macrophages at both the mRNA and the protein levels and significantly up-regulated MHC class II expression. Supernatants of α-toxin-stimulated macrophages induced the migration of human CD4+ lymphocytes via the CXCL10 receptor (CXCR3). Macrophages from patients with AD produced lower levels of CXCL10 compared to cells from patients with psoriasis as well as healthy controls in response to α-toxin. α-Toxin did not lead to a large variation in CCL22 production in macrophages from all three groups. CONCLUSIONS: Staphylococcal α-toxin contributes to Th1 polarization by induction of CXCL10 in macrophages. Macrophages from patients with AD and psoriasis responded to α-toxin in the induction of Th1-related chemokine CXCL10 diversely, which could favour the recruitment of distinct leucocyte subsets into the skin.

Interleukin (IL)-31 induces pro-inflammatory cytokines in human monocytes and macrophages following stimulation with staphylococcal exotoxins.

BACKGROUND: IL-31 is a cytokine expressed by T cells following activation with cytokines or staphylococcal exotoxins. A major function of IL-31 in atopic dermatitis (AD) is the induction of pruritus in the skin via the IL-31 receptor on sensory nerve cells. However, the regulation of the IL-31 receptor and pro-inflammatory functions of IL-31 in human monocytes and monocyte-derived cells are yet to be studied in detail. OBJECTIVE: To investigate the regulation and function of IL-31 receptors in resting and activated human monocytes, macrophages and dendritic cells. METHODS: Human monocytes, macrophages and dendritic cells were stimulated with staphylococcal exotoxins (SEB, alpha-toxin) or cytokines (IFN-gamma, IL-13). IL-31RA expression and regulation were then investigated at both the mRNA and the protein level. Subsequently, functional effects of IL-31 stimulation on cytokine secretion were measured at the protein level. RESULTS: Staphylococcal exotoxins significantly up-regulated IL-31RA expression on monocytes and macrophages but not on dendritic cells at both the mRNA and the protein level. IL-31 enhanced the secretion of IL-1beta, IL-6 and IL-18 and up-regulated CD86 expression. In patients with AD, functional IL-31RA was also detected following stimulation of PBMC with IFN-gamma. However, this was not observed in healthy individuals. CONCLUSION: IL-31 induces pro-inflammatory effects in activated human monocytes and macrophages. This may have implications for cutaneous inflammation in eczema where an over-expression of IL-31 has been described previously. Moreover, our findings provide a new link between staphylococcal colonization and the worsening of inflammation via IL-31. Further therapeutic considerations may include IL-31 as a target in AD.

Microleakage of dual-cured adhesive systems in class v composite resin restorations.

OBJECTIVE: Microleakage is a major factor affecting longevity of composite restorations. This study evaluated the effect of polymerization mode of bonding agent on microleakage of composite restorations. MATERIALS AND METHODS: Forty-eight Class V cavities were prepared on buccal and lingual surfaces of 24 extracted human premolars. Occlusal and gingival margins were placed in the enamel and dentin, respectively. Teeth were divided into four groups as follows: Group I: Optibond Solo Plus (light-cured); Group II: Optibond Solo Plus (dual-cured); Group III: Prime & Bond NT (light-cured), Group IV: Prime & Bond NT (dual-cured). Teeth were restored using Z250 composite in three increments. After polishing the restorations, samples were thermocycled for 1000 cycles and stored in distilled water for 3 months. Then they were placed in 2% fuchsine solution for 48 hours. The samples were sectioned longitudinally and evaluated for microleakage under a stereomicroscope at ×40 magnification. Dye penetration was scored on a 0-3 ordinal scale. Data were analyzed using Kruskal-Wallis, Bonferroni and Wilcoxon signed ranks test. RESULTS: Microleakage was significantly lower in enamel margins compared to dentin margins (P<0.05); multiple comparisons by Bonferroni tests revealed that the only factor with significant effect on leakage of the restoration is location of the restoration margin. Mode of adhesive polymerization had no significant influence on microleakage (P>0.05). Prime & Bond NT had less microleakage compared to Optibond SoloPlus, but the difference was not significant (P>0.05). CONCLUSION: There was no difference in the amount of microleakage in Class V composite restorations using light-cured and dual-cured bonding systems. Dentinal margins of restorations exhibited more microleakage than enamel margins.