EAMG is a B-cell-mediated, T-cell-dependent autoimmune disease where both-cell types play critical roles in disease progression and pathogenesis. To further define the primary target of the A2AR

agonist, cellular proliferation and anti-AChR antibody secretion of B cells were assessed in the presence of the A2AR agonist CGS21680 (30 nM). There were no significant differences in anti-AChR IgG production or in B-cell proliferation (Fig. 5), suggesting that CGS21680 had little effect on B-cell function. The effect of CGS21680 was next examined on CD4+ Th cells by examining the distribution of Th subsets following stimulation of cells with CGS21680 harvested from EAMG and CFA animals. This analysis demonstrated a decrease in the Th1 (CD4+/IFN-γ+; p < 0.05), Th2 (CD4+/IL-4+; p < https://www.selleckchem.com/products/Romidepsin-FK228.html 0.01), and Th17 (CD4+/IL-17+; p < 0.05) subsets and an increase in the Treg-cell subset (CD4+/CD25+/Foxp3+; p < 0.01) (Fig. 6A and B) after CGS21680 treatment of animals in the EAMG group compared with that of controls not treated with CGS21680 in vitro. Selleckchem Proteasome inhibitor To further confirm the Th profile observed, a cytokine profile analysis was carried out demonstrating that the concentration of IFN-γ (p < 0.001), IL-4 (p < 0.05), and IL-17 (p < 0.05) were

significantly decreased in the presence of CGS21680 (Fig. 6C). The concentration of IFN-γ was most significantly reduced. In contrast, TGF-β concentrations were significantly elevated (p < 0.01). These results suggested that A2AR activation downregulated Th1, Th2, and Th17 responses and promoted the development of the Treg-cell subset. Amylase To determine whether treatment of rats presenting with EAMG with CGS21680 could protect against disease progression, rats were given preventive treatment. EAMG symptoms were scored on alternate

days until the end of each experiment by assigning clinical scores and measuring changes in weight (Fig. 7A and C). Based on the ability of CGS21680 to significantly alter the clinical presentation of EAMG (p < 0.001), we next assessed the therapeutic potential of CGS21680 in animals with established EAMG. Rats treated with CGS21680 developed less weakness than rats in the EAMG group and elevated the clinical presentation and weight loss scores significantly (p < 0.05) (Fig. 7B and D). However, therapeutic treatment was not as efficient as preventive treatment. The efficacy of the CGS21680-preventive treatment was confirmed by evaluating the levels of anti-AChR IgG in serum and by measuring lymphocyte proliferation in ex vivo. Anti-AChR IgG titers and lymphocyte proliferation were significantly reduced in EAMG animals in the preventive (p < 0.001) (Fig. 8A and C) and therapeutic treatment group (p < 0.05) (Fig. 8B and D); however, therapeutic treatment was not as efficient as preventive treatment. Preventive CGS21680 administration was associated with a reduction in the proportion of Th1 (p < 0.01), Th2 (p < 0.05), and Th17 (p < 0.05) cells and an increase in Treg cells (p < 0.05) (Fig.