Immunogenic effects of recombinant interferon-beta therapy disrupt the JAK/STAT pathway in primary immune cells from patients with multiple sclerosis
Immunogenic effects of recombinant interferon-beta therapy disrupt the JAK/STAT pathway in primary immune cells from patients with multiple sclerosis
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DOI：
10.1177/1352458511434066
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发表时间：
2012-08-01
影响因子：
5.8
通讯作者：
Vedeler, C.
中科院分区：
文献类型：
--
作者：
Gavasso, S.;Gjertsen, B. T.;Vedeler, C.
背景：重组β干扰素的免疫原性是复发缓解型多发性硬化症(RRMS)治疗中的一个已知并发症。可干扰疗效的中和抗体(NAB)使用体外生物测定进行量化；目的：用磷酸化特异性流式细胞术评价NABS对外周血单核细胞(PBMC)中干扰素-β反应细胞和信号通路的影响。方法：10例经干扰素-β治疗的RRMS患者、两名未经治疗的患者和两名健康对照的PBMC在含有系列稀释度(0-8000U/ml)的干扰素-β(0-8000U/ml)的自体血清和培养液中再次刺激，并在PBMC亚型(NAB滴度0-GT；6000个中和单位)。数据经主成分分析、Hotling‘s T-2和偏最小二乘分析。结果：在自体血清中发现了三个显著不同的个体：治疗未成熟且健康的、治疗NAB阴性的和治疗NAB阳性的。与对照组相比，在NAB阴性的治疗患者中，STATS信号模式被调制，而在所有NAB阳性患者中，信号模式被抑制，而与NAB滴度无关。在媒体中，没有发现患者聚集。基于磷酸盐流动数据的NAB滴度的预测性为74%。结论：磷酸盐特异的流式细胞术可以描绘出亚群特异性的细胞反应，可以作为血液中NAB暴露的替代。即使在较低的NAB水平下，免疫原性效应也会改变原代细胞的反应。基于细胞系的免疫原性检测不容易转移到患者的免疫原性反应中。
Background: Immunogenicity of recombinant interferon-beta (IFN-beta) is a known complication in the therapy of relapsing-remitting multiple sclerosis (RRMS). Neutralizing antibodies (NAbs) that can interfere with efficacy are quantified using in vitro bioassays; however, these assays do not reveal the immunogenic state of the patient and are not predictive of treatment outcome.Objective: Assessment of the impact of NAbs on IFN-beta responsive cells and signalling pathways in peripheral blood mononuclear cells (PBMCs) with phospho-specific flow cytometry.Method: PBMCs from 10 IFN-beta-treated patients with RRMS, two untreated patients, and two healthy controls were re-stimulated in autologous sera and media with a serial dilution of IFN-beta (0-8000 U/ml) and levels of phosphorylation of STAT1/3/4/5/6 transcription factors were quantified in PBMC subtypes (NAb titres 0 to > 6000 neutralizing units). Data was subjected to principal component analysis, Hotelling's T-2, and partial least squares analysis.Results: Three significantly distinct clusters of individuals were revealed in autologous sera: therapy-naive and healthy, treated NAb-negative, and treated NAb-positive. Compared with controls STATs signalling patterns were modulated in treated NAb-negative patients and inhibited in all treated NAb-positive patients independently of NAb titres. In media no clustering of patients could be found. The predictability of NAb titres based on the phospho-flow data was 74%.Conclusion: Phospho-specific flow cytometry can delineate subset-specific cell responses that can act as surrogates for NAb exposure in blood. Immunogenic effects alter the response in primary cells even at low NAb levels. Cell line-based immunogenicity testing is not readily transferable to the immunogenic response in patients.