TY - JOUR
T1 - Comparison of multiplex platforms for cytokine assessments and their potential use for biomarker profiling in multiple sclerosis
AU - Malekzadeh, Arjan
AU - Twaalfhoven, H.A.M.
AU - Wijnstok, Nienke J.
AU - Killestein, Joep
AU - Blankenstein, Marinus A.
AU - Teunissen, Charlotte E.
PY - 2017/3/1
Y1 - 2017/3/1
N2 - Background 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. Methods 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. Results 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. Conclusion 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.
AB - Background 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. Methods 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. Results 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. Conclusion 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.
KW - CSF
KW - Multiple sclerosis
KW - Multiplex immunoassays
KW - Pro-inflammatory cytokines
KW - Serum
UR - http://www.scopus.com/inward/record.url?scp=85008879542&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.cyto.2016.12.021
DO - https://doi.org/10.1016/j.cyto.2016.12.021
M3 - Article
C2 - 28082233
SN - 1043-4666
VL - 91
SP - 145
EP - 152
JO - Cytokine Plus
JF - Cytokine Plus
ER -