|Immune phenotype (flow cytometry)||3 ml EDTA anticoagulated whole blood. The whole blood is analyzed on a Navios flow cytometer (Beckman-Coulter) within 24 h after blood sampling.||Proportion and intensity (mean fluorescence intensity, MFI) of antigens (most designated clusters of differentiation, CD) on the blood immune cells investigated in the flow cytometry panel.|
|Immune function (TruCulture®)||9 ml lithium heparin anticoagulated whole blood. The whole blood is transferred to individual TruCulture® tubes 60 min after blood sampling. After 22 h incubation at 37 °C, the TruCulture® supernatant is harvested and aliquoted to cryo tubes and frozen at − 20 °C for later thawing and bulk analysis of soluble immune activation products. The mRNA in the TruCulture® cell pellet is stabilized by Trizol and frozen at − 80 °C for later bulk analysis of mRNA expression level of stimulated immune proteins.||
TNF-α, IL-1β, IL-6, IL-8/CXCL8, IL-10, IL-12p40, IL-17A, IFN-γ|
Expression levels of the immune protein mRNA included in the multiplex assay will be calculated and reported.
|Circulating plasma biomarkers||6 ml EDTA and 3.5 ml sodium citrate 3.2% anticoagulated whole blood. The whole blood is spun (3000 RPM, 10 min) within maximum 6 h after blood sampling and plasma is aliquoted into cryo tubes and frozen at − 20 °C, and later transferred to − 80 °C. Plasma is thawed before analysis by Luminex®, MSD® or ELISA.||Circulating plasma levels of immune or inflammatory cytokines and autoantibodies against cytokines, chemokines, growth factors, adhesion molecules and injury/death products reflecting the magnitude of immune cell- and/or tissue activation and/or injury in vivo.|