Luminex x 200 instrument

The murine serum expression levels of eight cytokines, including IL-1β, IL-6, IL-10, IL-17A, IL-33, and TNF-α were measured using a Luminex cytokine assay kit (LXSAMSM-06, R&D Systems, USA) and a Luminex X-200 instrument according to the manufacturer’s protocol (R&D Systems, USA).

Serum levels of 35 cytokines, including C-C motif ligand (CCL)2, CCL4, CCL11, CCL20, CCL25, CCL26, CX3CL1, CXCL8, CXCL11, ADAMTS13, adiponectin, α-2-macroglobulin, matrix metalloproteinase (MMP)3, insulin-like growth factor binding protein-1, TNF-α, IL-1ß, IL-2, IL-4, IL-6, IL-7, IL-10, IL-11, IL-12 p70, IL-13, IL-17, IL-18, IL-23, IL-27, IL-28A, IL-28B, IL-31, IL-33, IL-34, and IL-36ß, at baseline and week 2 in discovery cohort were measured using a Luminex cytokine assay kit (LXSAHM-35) and a Luminex X-200 instrument according to the manufacturer’s protocol (R&D Systems, Minneapolis, MN, USA).
Enzyme-linked immunosorbent assay (ELISA) was used to measure levels of the molecules selected after the analysis of Luminex results obtained in the discovery cohort. In cohort 1 and 2, serum MMP3 levels were measured using a quantitative sandwich enzyme immunoassay (R&D Systems) at baseline and week 2. The serum CCL2 concentration was measured at week 2 using an ELISA kit (Abcam, Cambridge, UK, Ab179886). Serum IL-7 levels were measured using an ELISA kit (R&D Systems). Other indicators, such as CRP, ESR, albumin and so on, were tested with standard institutional protocols by the laboratory physicians at the First Affiliated Hospital of Sun Yat-sen University.