Amelioration of Experimental autoimmune encephalomyelitis and DSS induced colitis by NTG-A-009 through the inhibition of Th1 and Th17 cells differentiation

Abstract

CD4+ T cells are the central for the mammalian adaptive immune system. Naïve CD4+ T cells mainly differentiate in to pro-inflammatory Th1, Th2 and Th17 cells upon antigenic stimulation. IFN-γ secreting Th1 cells and IL-17 secreting Th17 cells are found to play key roles in autoimmune diseases like multiple sclerosis (MS) and ulcerative colitis (UC). In this study we found NTG-A-009, 6-aminopyridin-3-ol, has great inhibitory effect on in vitro differentiation of Th1 and Th17 cells without affecting regulatory T cells. Moreover, NTG-A-009 had no effect on CD4+ T cell proliferation and viability. In vivo treatment has shown that NTG-A-009 has ameliorated experimental autoimmune encephalomyelitis (EAE) and dextran sulfate sodium (DSS) induced colitis through the inhibition of Th1 and Th17 cells differentiation. Mechanistically, NTG-A-009 suppressed Th1 and Th17 cells differentiation via the modulation of JAK/STAT signaling pathway. Thus, our data demonstrated that NTG-A-009 ameliorated inflammation through the inhibition of Th1 and Th17 cells generation making it a potential therapeutic candidate for the treatment of inflammatory diseases.

The Function and Role of the Th17/Treg Cell Balance in Inflammatory Bowel Disease

“…Compared with Th17 cells, Treg cells not only suppress the occurrence of autoimmune diseases but also control intestinal inflammation. In the UC mouse model, Treg cells in the peripheral blood of mice decreased [ 15 ], and by increasing the secretion of IL-10 and TGF- β , the symptoms of diarrhoea in mice were significantly improved [ 16 ]. Therefore, Treg cells may be regulated by IL-10, TGF- β , and other anti-inflammatory factors that are secreted to suppress the intestinal inflammation cascade and amplify the response, thereby improving the clinical symptoms of IBD.…”

Geranylgeranyl pyrophosphate amplifies Treg differentiation via increased IL‐2 expression to ameliorate DSS‐induced colitis

“…Isolated CD4 + CD25 − T cells from mouse spleen and LNs were stimulated with plate‐bound anti‐CD3 (145‐2CC21, BioLegend) (5 μg/mL) and anti‐CD28 (37.51, BioLegend) (1 μg/mL) under Th1, Th17, or T reg polarizing conditions as previously [49] in the presence of GGPP‐geranylgeraniol and ATRA. For, IL1‐β‐ and IL‐23‐induced in vitro Th17 differentiation, CD4 + CD25 ‐ T cells were stimulated with plate‐bound anti‐CD3 (5 μg/mL) and anti‐CD28 (1 μg/mL) under IL‐6 (20 ng/mL), IL‐1β (20 ng/mL), and IL‐23 (50 ng/mL) cytokines; all from R&D systems as described previously [50] in presence or absence of GGPP and ATRA.…”

“…Changes in body weight and bleeding scores were noted daily and histopathological analysis of colon was assessed. Bleeding score were measured on a scale of 0‐3 (0: normal stool; 1: slightly bloody with soft stool; 2: traces of blood with very soft stool; 3: gross bleeding with diarrhea) as described previously [49].…”

“…Cells were then separated using percoll gradient (40% overlaid on 70%). Lamina propria lymphocytes were collected from the interface as described previously [49]. For IFN‐γ and IL‐17 intracellular staining, isolated lymphocytes were stimulated with PMA and ionomycin along with protein transport inhibitors for 4 h and stained for flow cytometry analysis.…”

Differential regulation of JAK/STAT-signaling in patients with ulcerative colitis and Crohn’s disease

“…Furthermore, several anti-inflammatory molecules ameliorating disease activity in experimental colitis are associated with modulation of JAK/STAT-signaling in lymphocytes. In detail, decreased expression or activation of JAK1 and JAK2[ 77 ], STAT1[ 77 ], STAT3[ 77 - 79 ] and STAT4[ 77 , 79 ] as well as enhancement of STAT5[ 78 ] phosphorylation in T-cells, have been linked to effective pharmacological treatment of DSS-induced murine colitis and TNBS-induced rat colitis. Additionally, inflammatory activity was reduced by general STAT1 knockout in DSS-treated STAT1-null mice[ 80 ] and by specific inhibition of STAT1 in T-cells in TNBS-induced colitis[ 81 ].…”