UNASSIGNED: Serum titers of anti-IFN-γ autoantibodies were quantified using ELISA in 29 AOSD and 22 healthy controls (HC). The detectable autoantibodies were verified with immunoblotting assay, and their neutralizing capacity against IFN-γ-signaling was evaluated with flow-cytometry analysis and immunoblotting. IFN-γ-mediated production of supernatant chemokines, including monocyte chemoattractant protein-1 (MCP-1) and IFN-γ inducible protein-10 (IP-10), were measured by ELISA.
UNASSIGNED: Among 29 AOSD patients, high titers of anti-IFN-γ neutralizing autoantibodies were detectable in two patients with OIs. Immunoblotting assay revealed more effective inhibition of STAT1-phosphorylation in THP-1 cells treated with sera from autoantibody-positive AOSD patients (56.7 ± 34.79%) compared with those from HC (104.3 ±29.51%), which was also demonstrated in flow-cytometry analysis (47.13 ± 40.99 vs. 97.92 ± 9.48%, p < 0.05). Depleted serum IgG from anti-IFN-γ autoAbs-positive AOSD patients with OIs restored phosphorylated STAT-1 upon IFN-γ treatment. Sera from autoantibody-positive AOSD patients more effectively inhibited IFN-γ-mediated production of MCP-1 (45.65 pg/ml) and IP-10 (22.44 pg/ml) than sera from HC (263.1 pg/ml and 104.0 pg/ml, both p < 0.05). Serum samples showing the strongest inhibition of IFN-γ-signaling were from two patients with high-titer autoantibodies and OIs.
UNASSIGNED: AOSD patients have a high positive rate and titers of anti-IFN-γ autoantibodies. The remarkable blockade effect of high-titer autoantibodies on IFN-γ-mediated STAT1-phosphorylation and chemokines could make these patients susceptible to OIs.
UNASSIGNED:使用ELISA在29个AOSD和22个健康对照(HC)中定量抗IFN-γ自身抗体的血清滴度。用免疫印迹法验证可检测的自身抗体,通过流式细胞术分析和免疫印迹评估了它们对IFN-γ信号的中和能力。IFN-γ介导的上清液趋化因子的产生,包括单核细胞趋化蛋白-1(MCP-1)和IFN-γ诱导蛋白-10(IP-10),通过ELISA测量。
未经证实:在29名AOSD患者中,在两名OIs患者中检测到高滴度的抗IFN-γ中和自身抗体。免疫印迹分析显示,与HC(104.3±29.51%)相比,用自身抗体阳性AOSD患者的血清(56.7±34.79%)处理的THP-1细胞对STAT1磷酸化的抑制作用更有效,这也在流式细胞术分析中得到了证明(47.13±40.99vs.97.92±9.48%,p<0.05)。来自抗IFN-γ自体Ab阳性AOSD患者的血清IgG在IFN-γ治疗后恢复磷酸化STAT-1。来自自身抗体阳性AOSD患者的血清比来自HC的血清(263.1pg/ml和104.0pg/ml)更有效地抑制IFN-γ介导的MCP-1(45.65pg/ml)和IP-10(22.44pg/ml)的产生。两者p<0.05)。显示IFN-γ信号传导最强抑制的血清样品来自具有高滴度自身抗体和OI的两名患者。
UASSIGNED:AOSD患者的抗IFN-γ自身抗体阳性率和滴度较高。高滴度自身抗体对IFN-γ介导的STAT1磷酸化和趋化因子的显着阻断作用可能使这些患者对OIs敏感。