Luminex is 100 instrument

Soluble (s)CD163 was analyzed in INRs and immunological responders from snap frozen EDTA plasma in duplicate by immunoassay (DY1607; R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer's instructions. The intraassay and interassay coefficients of variation (CV) were less than 10%.
Concentrations of IL-1β, IL-1 receptor antagonist (IL-1ra), IL-6, IL-10, IL-18, IFNα2, IFNγ, tumor necrosis factor, IP-10 and macrophage inflammatory protein 1beta (MIP-1β)/CCL4 were measured in thawed supernatants (10 000 g/10 min/4 °C) by nine-plex and single-plex (Bio-Rad, Oslo, Norway) assays, respectively. The samples were diluted 1 : 7.5 and 1 : 75 as appropriate, analyzed with a Luminex IS 100 instrument (Bio-Rad, Hercules, California, USA) according to instructions from the manufacturer and run in duplicates. Both intraassay and interassay CV were less than 10%. All AT-2 HIV virus responses were found by subtracting concentrations from the corresponding mock stimulated sample.
Multiplex and ELISA analyses of other soluble markers in plasma and flow cytometry phenotyping of T-cell subsets including Tregs have been previously presented [16 (link)], and are only used in correlation analyses in the current article. The methods are described in brief in the Supplementary methods.

Duplicate serum analysis by a 40-plex Pro Human Chemokine multi-bead assay (Bio-Rad, Norway) was used to assess the concentrations of 40 cytokines in patients and control subjects (table 1). The panel was chosen as it included cytokines that previously has been linked to MCs as interleukin (IL)1-β), tumor necrosis factor alpha (TNF-α), C-X-C Motif Chemokine Ligand 5 (CXCL5), granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-6, IL-8 and macrophage migration inhibitory factor (MIF).18 20 27 28 (link) Data were recorded with a Luminex IS 100 instrument (Bio-Rad, Hercules, California, USA) and protein concentrations were determined using recombinant standard curves. The following cytokines were analysed: (TNF-α), interferon gamma (IFN-γ), IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-16, GM-CSF, MIF, C-C motif ligand(CCL)1, CCL2, CCL3, CCL7, CCL8, CCL11, CCL13, CCL15, CCL17, CCL19, CCL20, CCL21, CCL22, CCL 23, CCL24, CCL25, CCL26, CCL27, CXCL1, CXCL2, CXCL5, CXCL6, CXCL9, CXCL10, CXCL11, CXCL12, CXCL13, CXCL16 and C-X3-C Motif Chemokine Ligand 1 (CX3CL1). Only cytokines with less than 20% missing values were included in further analyses. CXCL5 was below the limit of quantification for more than half of the samples and excluded from further analysis.

Citrated whole blood was prepared by centrifugation at 2500 gfor 15 min at RT. Then, 1 ml plasma was collected in a 1.5-ml Eppendorf tube and
centrifuged at 10,000 g for 10 min at 4℃. The top supernatant
(800 µl) was collected prior to freezing, and stored at –80℃. Measurements of
cytokines were performed using a microsphere-based multiplexing bioassay system
with Xmap technology (Austin, TX, USA) with a Luminex IS 100 instrument
(Bio-Rad, Hercules, CA, USA), powered with the Bio-Plex manager Software version
6.0 (build 617). The anti-inflammatory cytokine IL-10, the MyD88-dependent
cytokines IL-1β, IL-6, IL-8 and TNF-α,¹³ and the MyD88-independent
cytokines IP-10 and RANTES,^{15 (link)} were measured (Bio-Rad, Oslo, Norway) in duplicates.