Malekzadeh Arjan, Twaalfhoven Harry, Wijnstok Nienke J, Killestein Joep, Blankenstein Marinus A, Teunissen Charlotte E
Department of Clinical Chemistry, VU University Medical Center, Amsterdam, The Netherlands.
Department of Internal and Vascular Medicine, VU University Medical Center, Amsterdam, The Netherlands.
Cytokine. 2017 Mar;91:145-152. doi: 10.1016/j.cyto.2016.12.021. Epub 2017 Jan 9.
The levels of pro and anti-inflammatory cytokines can be altered in different autoimmune pathologies, such as multiple sclerosis (MS). It is likely that cytokines in bodily fluids can provide a good reflection of ongoing disease patho-physiology. In this study we aimed to validate multiplex cytokine platforms and evaluate whether these cytokines are differentially expressed in MS.
Assay validation for simultaneous quantification of IL-1β, IL-6, IL-8 and TNF-α in serum and CSF were performed using both the Luminex-xMAP (Luminex) and Meso Scale Discovery (MSD) platforms. Next, the relation of the pro-inflammatory cytokine 4-plex with disease progression, symptoms and subtypes was studied in paired serum and CSF of MS patients (n=56), and compared with healthy controls (n=203), with the use of the MSD-platform.
The MSD-platform showed overall better assay characteristics such as, sensitivity, recovery and linearity compared to the Luminex for the 4-plex cytokines in CSF and serum. IL-6, IL-8 and TNF-α (p<0.001) levels were significantly increased in MS serum compared to healthy controls. Moreover, serum IL-1β levels correlated with expanded disability status scale (EDSS) scores (r=-0.34, p<0.05). Additionally, IL-6 and IL-8 CSF levels were both significantly decreased in MS patients compared to non-inflammatory neurological disease controls. Noteworthy, higher IL-8 CSF levels than IL-8 serum levels were observed for MS patients, indicating intrathecal activation of macrophages in MS.
We have demonstrated that the pro-inflammatory 4-plex kit of the MSD-platform shows better assay characteristics in comparison with Luminex kit for quantification of these cytokines in serum and CSF. Overall, the increased levels of IL-6, IL-8 and TNF-α in serum of MS patients compared to healthy controls, support the use of multiple cytokines for future MS biomarker and disease progression research.
促炎和抗炎细胞因子的水平在不同的自身免疫性疾病中可能会发生改变，如多发性硬化症（MS）。体液中的细胞因子很可能能够很好地反映正在进行的疾病病理生理学情况。在本研究中，我们旨在验证多重细胞因子检测平台，并评估这些细胞因子在MS中是否存在差异表达。
使用Luminex-xMAP（Luminex）和Meso Scale Discovery（MSD）平台对血清和脑脊液中白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、白细胞介素-8（IL-8）和肿瘤坏死因子-α（TNF-α）的同时定量进行检测验证。接下来，使用MSD平台，在MS患者（n=56）的配对血清和脑脊液中研究促炎细胞因子四联检测与疾病进展、症状和亚型的关系，并与健康对照（n=203）进行比较。
与Luminex相比，MSD平台在脑脊液和血清中对四联细胞因子的检测总体上表现出更好的检测特性，如灵敏度、回收率和线性。与健康对照相比，MS血清中IL-6、IL-8和TNF-α（p<0.001）水平显著升高。此外，血清IL-1β水平与扩展残疾状态量表（EDSS）评分相关（r=-0.34，p<0.05）。此外，与非炎性神经疾病对照相比，MS患者脑脊液中IL-6和IL-8水平均显著降低。值得注意的是，MS患者脑脊液中IL-8水平高于血清中IL-8水平，表明MS中巨噬细胞的鞘内激活。
我们已经证明，与Luminex试剂盒相比，MSD平台的促炎四联检测试剂盒在血清和脑脊液中定量这些细胞因子时表现出更好的检测特性。总体而言，与健康对照相比，MS患者血清中IL-6、IL-8和TNF-α水平升高，支持将多种细胞因子用于未来MS生物标志物和疾病进展研究。
