A comparison of multiplex suspension array large‐panel kits for profiling cytokines and chemokines in rheumatoid arthritis patients

Abstract

Background Multiplex analysis allows measurements of a large number of analytes simultaneously in each sample. Based on the Luminex multiplex technology (xMAP), kits for measuring multiple cytokines and chemokines (immunomodulators) are commercially available and are useful in investigations on inflammatory diseases. This study evaluated four multiplex kits (Bio-Plex, LINCOplex, Fluorokine, and Beadlyte) that contained 27, 29, 20 and 22 analytes each, respectively, for the analysis of immunomodulators in plasma of rheumatoid arthritis (RA) patients who underwent treatment with antibody against CD20 (rituximab), a B-cell reductive therapy. Methods Multiplex kits were tested on serial plasma samples obtained from six RA patients at baseline and multiple time points (3, 6, and 9 months) post-treatment with rituximab. The RA patients included in this study had previously failed therapy with disease modifying anti-arthritis drugs (DMARD) and treatment with anti-TNFα antibody (infliximab). Results Computer modeling and hierarchical cluster analysis of the multiplex data allowed a comparison of the performance of multiplex assay kits and revealed profiles of immunomodulators in the RA patients. Conclusions In plasma of RA patients who appeared to have benefited from rituximab treatment the profile of significantly elevated immunomodulators by at least two of the three kits (BioPlex, LINCOplex, Beadlyte), is as follows: IL-12p70, Eotaxin, IL-4, TNFα, Il-9, IL-1β, IFNγ, IL-10, IL-6, and IL-13. Immunomodulator profiling by multiplex analysis may provide useful plasma biomarkers for monitoring response to B-cell reductive therapy in RA patients.

Increased interleukin-9 and CD4+IL-9+ T cells in patients with systemic lupus erythematosus

“…Poulin et al (14) found that IL-9 mRNA and protein were highly expressed in a mouse model of fully mismatched heart transplantation. A previous study also demonstrated that serum levels of IL-9 and additional pathologic cytokines were elevated after the second immunization of rheumatoid arthritis (RA) (15), while Khan et al (16) further confirmed that IL-9 and some another inflammatory cytokines were descended following treatment with rituximab, indicating that IL-9 plays a pro-inflammatory role in RA. Additional evidence indicates that IL-9 cytokine may contribute to the development of autoimmune diseases.…”

Exploratory Study on Plasma Immunomodulator and Antibody Profiles in Tuberculosis Patients

“…A linear model fit was determined for each analyte by using the LIMMA package in R. The data corresponding to immunomodulators (and antibodies) with a positive signal were combined in a two-step process to obtain an initial analytical data set. Relative fold changes (increase or decrease) of the levels of both immunomodulators and antibodies between the healthy controls and patient groups in the pilot as well as the expanded experiments were calculated according to a previously described procedure (19). Briefly, fold changes were calculated across conditions by fitting a linear model to the data, and the statistical significance was estimated by a two-step process.…”

“…Immunomodulators were selected based on the fold change as well as adjusted P values for multiple comparisons. Immunomodulators with a fold change of at least 2 and a P value of Ͻ0.01 were considered significant (19).…”

Multiplex Bead Assay for Serum Samples from Children in Haiti Enrolled in a Drug Study for the Treatment of Lymphatic Filariasis

“…[1][2][3][4][5][6][7][8][9][10] The popularity of the MBA is the result of 1) the relative ease of covalently coupling of an analyte-capture ligand to spectrally classified carboxylated microspheres; 2) the simultaneous collection of multiple data points from a single specimen, eliminating the one-data point per well enzyme-linked immunosorbent assay (ELISA); 3) the direct proportionality of the fluorescence intensity of the reporter fluorochrome to the amount of captured analyte; and 4) the use of a 96-well format with up to 100 differently classified beads per well, which yield a potential of 9,600 data points per plate. These attractive features of the MBA conserve specimens and save on labor, time, and cost when compared with the ELISA format.…”