Changes of the expression of IL-4 and IFN-γ in monocytes and T-helper lymphocytes while modeling of systemic juvenile idiopathic arthritis in Wistar rats
Abstract
The objective of the study was to assess the opportunities of targeted reprogramming of peripheral blood mononuclear cells in a model of systemic juvenile idiopathic arthritis (sJIA) in Wistar rats (based on combined administration of complete Freund's adjuvant and lipopolysaccharide). Methods. The modeling of sJIA was performed on 6-month-old male Wistar rats. The animals were divided into three groups (on day 40): control, doxycycline and dexamethasone ones. Blood samples were harvested on days 0, 41 and 55. Peripheral blood mononuclear cells were obtained gravimetrically and then stained for markers and cytokines. As targeted cells monocytes (CD3-CD4+) and T-helpers (CD3+CD4+) were distinguished. Flow cytometric analysis of intracellular expression (and its dynamics) of IL-4 (regarded as marker of pro-M2 phenotype since IL-4 after releasing can activate M2 macrophage polarization) and IFN-γ (regarded as marker of pro-M1 phenotype) was performed. Nonparametric statistical tests (Mann-Whitney-Wilcoxon) were performed in R environment for statistical computing. Results and conclusions. Monocytes (just like macrophages) underwent reprogramming in model sJIA disease course. In groups with additional doxycycline or dexamethasone administration pro-M2 effects in monocytes were observed earlier (day 41) than in control group. Pro-M1 effects in monocytes were observed in doxycycline and dexamethasone groups on day 55 (in comparison with control group). Also the characteristic dynamical changes of intracellular cytokine expression in animal groups are described. Importantly, observed changes confirm obtained different cell reprogramming (for both monocytes and T-helpers) in animal groups.
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