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. 2015 Sep 15;8(9):16158-66.
eCollection 2015.

Performance evaluation of FlowCytomix assays to quantify cytokines in patients with rheumatoid arthritis

Xuefeng Wang  1 , Liyang Dong  2 , Yong Liang  3 , Hongchang Ni  4 , Jun Tang  4 , Chengcheng Xu  2 , Yuepeng Zhou  2 , Yuting Su  2 , Jun Wang  4 , Deyu Chen  2 , Chaoming Mao  2
Affiliations

Performance evaluation of FlowCytomix assays to quantify cytokines in patients with rheumatoid arthritis

Xuefeng Wang et al. Int J Clin Exp Med. .

Abstract

Objectives: To compare the cytokine profile in RA patients and healthy control by using two methods-FlowCytomix assay and traditional ELISA.

Methods: Cytokine levels were evaluated by FlowCytomix assay and ELISA in serum and supernatants of peripheral blood mononuclear cells (PBMC) cultures with and without stimulation by phytohaemagglutinin (PHA).

Results: The levels of IL-6, IL-1β, and TNF-α were significantly higher in sera of RA patients than those of healthy controls. The levels of IL-22, IL-6, IL-1β, TNF-α, and IL-10 were higher in unstimulated PBMC culture supernatant of RA patients than those of healthy controls. PHA stimulation significantly increased the production of proinflammatory cytokines from PBMC with RA patients. Compared with detectable cytokine levels in sera, cytokine concentration in the supernatant of PBMCs was remarkably higher. FlowCytomix and ELISA showed significant correlation in detecting cytokines. However, the FlowCytomix assay detected more cytokines than ELISA.

Conclusion: The supernatant of PBMCs provide a fine condition for the study of cytokine production because of the lack of interference factors in sera. The FlowCytomix assay is more sensitive than ELISA in detecting cytokines from RA patients. Multiple cytokine signatures using FlowCytomix assay may represent a more realistic approach in the future of personalized medicine in RA.

Keywords: Cytokine profile; ELISA; FlowCytomix assay; rheumatoid arthritis.

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Figures

Figure 1
Figure 1
Correlation of cytokines in supernatants of PHA-stimulated PBMCs from RA patients detected between FlowCytomix and ELISA. Correlation of pg/ml of IFN-γ (A), IL-17 (B), IL-22 (C), IL-6 (D), IL-1β (E), TNF-α (F), and IL-10 (G) detected between the FlowCytomix and ELISA. The two methods significantly correlated with one another for the detection of IFN-γ (Pearson r = 0.88, P = 0.004), IL-17 (Pearson r = 0.94, P = 0.0005), IL-22 (Pearson r = 0.92, P = 0.0014), IL-6 (Pearson r = 0.89, P = 0.0031), IL-1β (Pearson r = 0.95, P = 0.0003), TNF-α (Pearson r = 0.97, P < 0.001), and IL-10 (Pearson r = 0.94, P =0.0006).
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