Cytokine Profiling in Plasma from Patients with Brain Tumors Versus Healthy Individuals using 2 Different Multiplex Immunoassay Platforms

Abstract

We compared the performance of two 96-well multiplex immunoassay platforms in assessing plasma cytokine concentrations in patients with glioblastoma (GBM; n = 27), individuals with melanoma, breast or lung cancer metastases to the brain (n = 17), and healthy volunteers (n = 11). Assays included a bead-based fluorescence MILLIPLEX® assay/Luminex (LMX) platform and 4 planar electrochemiluminescence kits from Meso Scale Discovery (MSD). The LMX kit evaluated 21 cytokines and the 3 MSD kits evaluated 20 cytokines in total, with 19 overlapping human cytokines between platforms (GM-CSF, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-17A, IL-21, IL-23, MIP-1α, MIP-1β, MIP-3α, TNFα). The MSD platform had lower LLoQs (lower limits of quantification) than LMX for 17/19 cytokines, and higher LLoQs for IFN-γ and IL-21. The ULoQs were higher in LMX versus MSD assays for 17/19 shared analytes, but lower than MSD for IL-17A and IL-21. With LMX, all 19 shared analytes were quantifiable in each of 55 samples. Although MSD recombinant protein standard curves indicated lower LLoQs than LMX for most cytokines, MSD detected 7/19 (37%) native analytes in <75% of samples, including 0% detection for IL-21 and 8% for IL-23. The LMX platform categorized identical samples at greater concentrations than the MSD system for most analytes (MIP-1β the sole exception), sometimes by orders of magnitude. This mismatched quantification paradigm was supported by Bland-Altman analysis. LMX identified significantly elevated levels of 10 of 19 circulating cytokines in GBM: GM-CSF, IFN-γ, IL-1β, IL-5, IL-10, IL-17A, IL-21, IL-23, MIP-1α, and MIP-3α, consistent with prior findings and confirming the utility of applying appropriate multiplex immunoassay technologies toward developing a cytokine signature profile for GBM.

A phase 1b/2a clinical trial of dantrolene sodium in patients with Wolfram syndrome

“…Therefore, we assessed expression levels of inflammatory cytokines and isoprostane, an established marker of oxidative stress in study subjects prior to and 6 months after starting dantrolene treatment via WURS assessment (34-36) (Table 4). As compared to levels previously reported in heathy subjects (35)(36)(37), levels of INF, TNF, IL-1, MIP-1, MIP-3, IL-21, and isoprostane are higher in study subjects with Wolfram syndrome. Among those, a statistically…”

Risk assessment of drug–drug interaction potential for bintrafusp alfa with cytochrome P4503A4 substrates: A totality of evidence approach

“…Based on analyses conducted on samples from a phase I study in patients with metastatic or locally advanced solid tumors, bintrafusp alfa did not cause clinically relevant modulation of proinflammatory cytokines implicated in CYP3A4 enzyme modulation (Figure 1a). In this data set, baseline cytokine or 4β‐hydroxycholesterol levels (Tables S2 and S3) appeared to overlap with the ranges reported in healthy patients 13,20,21 . The only cytokine with evidence of transient (2.4‐fold) increase was IFN‐ɣ, which had decreased toward the baseline by 2 weeks after the dose.…”

“…In this data set, baseline cytokine or 4β‐hydroxycholesterol levels (Tables S2 and S3 ) appeared to overlap with the ranges reported in healthy patients. 13 , 20 , 21 The only cytokine with evidence of transient (2.4‐fold) increase was IFN‐ɣ, which had decreased toward the baseline by 2 weeks after the dose. No modulation of any other inflammatory cytokines was observed with bintrafusp alfa.…”

Effects of Distinct Force Magnitude of Spinal Manipulative Therapy on Blood Biomarkers of Inflammation: A Proof of Principle Study in Healthy Young Adults

“…In addition, reference concentration (mean § SD) from previous studies using similar individuals (healthy adults) and similar sample-related technical features (sample type: plasma; anticoagulant: EDTA; assay: bead-based fluorescence multiplex assay; similar commercial kit) were included in Table 2 to facilitate external comparison. 31,[35][36][37][38] No statistical difference in each plasmatic cytokine concentration among the 3 groups was observed at the baseline (P ≥ .05). Therefore, the change score from baseline could be calculated.…”