AB0012 THE EFFECTS OF COMMON THERAPEUTICS ON AIRE METHYLATION STATUS AND THE LEVELS OF IL-16, IL-1Β, AND IFN-Γ IN LPS-INDUCED MACROPHAGE CELLS CAN SULPHASALAZINE’S INFLUENCE ON METHYLATION STATUS OF AIRE EXPLAIN ITS AUTOIMMUNE SIDE EFFECTS?

Background: The autoimmune regulator gene ( AIRE ) has very important role in self-tolerance. Estrogen induced hypermethylation of promoter was put forward as one the reasons for susceptibility to autoimmunity females. [1] Sulphasalazine is a commonly used drug rheumatology and known be responsible various side effects. Objectives: We aimed investigate effects treatments on methylation status levels interleukin-16 (IL16), interleukin-1β (IL1β), interferon-γ (IFNγ), lipopolysaccharides (LPS)-induced RAW264.7 macrophage cells which mimic inflammatory state rheumatoid arthritis vitro.[2] Methods: were stimulated by LPS (1μ/mL). Cell viability test performed determine concentrations. Following treatments, cell media isolated determination IL16, IL1β, IFNγ ELISA. Also, detached trypsin-EDTA DNA isolation bisulfite modification. Then, analyzed with methylation-specific PCR agarose jel electrophoresis. Results: Our results demonstrated that highly methylated absence any stimulus.LPS treatment changed unmethylated form. Leflunomide(LEF), sulfasalazine(SLZ) methotrexate(MTX) significantly altered (p=0.041,p<0.001,p<0.001, respectively). Surprisingly, SLZ caused more than other drugs (p<0.001,Fig 1A). Analysis IL1β potent increase all three cytokines (p<0.001, Fig 1B-D). LEF prevented LPS-induced these (p=0.025, p<0.001 IL-16, IL-1β IFNγ, Although MTX exacerbated IL16 levels, it inhibited Figure 1. gel image. All data expressed mean±SD. ***p<0.001 versus control, ###p<0.001, #p<0.05 LPS, +++p<0.001 sulfasalazine. Conclusion: suggest although LEF,SLZ potently suppresses cytokines, they affect differently.Changes are autoimmunity. Relative usage can its effects, like lupus or hypersensitivity reactions, not common usage. key biological properties CD4+T cells, also regulates migration CD4+CD25+Treg cells.[3] shown Treg early patients. SLZ’s effect different from MTX.[4] contrary SLZ. This difference may medications functions. disease modifying interesting deserves attention. References: [1]Berrih-Aknin, Le Panse R, Dragin N. AIRE:a missing link explain female diseases. Ann.N.Y.Acad.Sci, 2018;1412(1):21-32. [2]Lin Y-Y, Jean Y-H, Lee H-P et al. Excavatolide B attenuates through inhibition osteoclastogenesis. Marine Drugs, 2017; 15(1):9. [3]Skundric DS, Cruikshank WW, Montgomery PC Emerging IL-16 cytokine-mediated regulation multiple sclerosis. Cytokine, 2015;75(2):234-248. [4]Oh JS, Kim Y-G, SG anti-rheumatic suppressive function CD4+CD25+ regulatory Tcells. Rhematol Int. 2013;33:381-388. Disclosure Interests: None declared