Using serum metabolomics to predict development of anti-drug antibodies in multiple sclerosis patients treated with IFNβ

Hypothesis: Serum metabolomics can be used to predict immunogenicity against IFNβ in multiple sclerosis patients A prospective cohort of multiple sclerosis (MS) patients was recruited across six European countries as part of the Anti-Biopharmaceutical Immunization: prediction and analysis of clinical relevance to minimize the RISK consortium (ABIRISK consortium; www.abirisk.eu/). Serum samples were collected from MS patients prior to IFNβ treatment (M0), and after 3 (M3) and 12 (M12) months to be used for anti-drug antibody detection (ADA) and metabolomic analysis. 228 serum metabolites and lipids were quantified using an established nuclear magnetic resonance spectroscopy platform (Nightingale Health). The dataset includes absolute concentrations (mmol), and relative measures (ratios, percentages) from 82 MS patients (M0 and M12). 9 were missing at M3, leaving n=73 at this time-point. Serum was tested for both binding (bAbs) and neutralizing (nAbs) ADA, measured with an enzyme-linked immunosorbent assay (ELISA) (Ingenhoven et al., 2017) and cell-based luciferase reporter gene assay (Hermanrud et al., 2016), respectively. Patients were classified as ADA positive if they were positive for bAbs, or had a nAbs titer >320 U/mL within 12 months of starting treatment. Patients were considered ADA negative if they were negative for both assays. Patients with missing data or negative for bAbs and with a nAbs titer <320 U/mL were excluded.研究假说：血清代谢组学可用于预测多发性硬化（multiple sclerosis, MS）患者对IFNβ的免疫原性。本研究作为"抗生物制剂免疫：临床相关性预测与分析以最小化风险联盟"（Anti-Biopharmaceutical Immunization: prediction and analysis of clinical relevance to minimize the RISK consortium，简称ABIRISK联盟；www.abirisk.eu/）的一部分，在六个欧洲国家招募了多发性硬化患者组成前瞻性队列。研究采集了多发性硬化患者接受IFNβ治疗前（M0）、治疗后3个月（M3）及12个月（M12）的血清样本，用于抗药物抗体（anti-drug antibody, ADA）检测与代谢组学分析。依托成熟的核磁共振波谱平台（Nightingale Health），共定量检测了228种血清代谢物与脂质。数据集包含82名多发性硬化患者在M0和M12时间点的绝对浓度（单位：mmol）及相对测量值（比值、百分比）；M3时间点存在9例样本缺失，该时间点有效样本量为n=73。血清样本分别接受结合抗体（binding antibodies, bAbs）与中和抗体（neutralizing antibodies, nAbs）类抗药物抗体检测：结合抗体采用酶联免疫吸附试验（enzyme-linked immunosorbent assay, ELISA；Ingenhoven等，2017）完成，中和抗体则采用基于细胞的荧光素酶报告基因试验（Hermanrud等，2016）完成。患者分类标准如下：若结合抗体检测阳性，或启动治疗后12个月内中和抗体滴度＞320 U/mL，则归类为ADA阳性；若两项检测均为阴性，则归类为ADA阴性。存在数据缺失、结合抗体检测阴性且中和抗体滴度＜320 U/mL的患者被排除出本数据集。