Blockade of interferon-γ normalizes IFN regulated gene expression in subjects with systemic lupus erythematosus (SLE)

Normal donor blood was incubated with or without IFN-g stimulation to establish an IFN-g gene signature. Systemic lupus erythematosus subjects were treated with placebo or AMG 811, a therapeutic anti-IFN--g antibody, and changes in the IFN--g signature in whole blood of these subjects was measured. Blood from healthy volunteers (n=4) was collected into sodium heparin tubes, and then untreated or treated with 294 pM recombinant human IFN-γ for 0, 24, or 48 hours with incubation at 37oC, 5 % CO2. The blood was then added to PAXgene tubes and processed for RNA purification. Twenty six subjects with stable, mild to moderate SLE were administered placebo or a single dose of AMG 811 ranging from 2 mg to 180 mg SC or 60 mg IV. Whole blood PAXgene tube samples were collected from all cohorts at baseline, day-1 (pre-dose), and at days 15, 56, and end of study (EOS) after treatment Arrays were hybridized in a Loop design.

本研究以健康供血者血液为实验材料，设置干扰素γ（IFN-γ）刺激与未刺激两组，以此构建IFN-γ基因特征谱。随后对系统性红斑狼疮（Systemic lupus erythematosus, SLE）受试者施以安慰剂或AMG 811（一种治疗性抗IFN-γ抗体）干预，并检测该类受试者全血中IFN-γ特征谱的变化情况。采集共4名健康志愿者的血液至肝素钠采血管（sodium heparin tubes）中，分别进行未处理与294 pM重组人IFN-γ处理，于37℃、5%CO₂培养条件下孵育0、24、48小时。随后将血液转移至PAXgene采血管（PAXgene tubes）中，进行RNA纯化相关处理。26名病情稳定、轻至中度的SLE受试者被分配接受安慰剂，或单次剂量的AMG 811干预：皮下注射（SC）剂量范围为2 mg至180 mg，静脉注射（IV）剂量为60 mg。在所有队列的基线期、给药前第1天、给药后第15天、第56天以及研究终点（EOS）采集全血PAXgene采血管样本。实验采用循环设计完成基因芯片阵列杂交。