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.
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.