Skip to main content

Table 1 Method and data output of the immunological profile. Description of blood sample collection, laboratory analysis and data-output from the immunological profile, consisting of immune phenotype (flow cytometry), immune function (TruCulture®) and circulating plasma biomarkers

From: Immune function as predictor of infectious complications and clinical outcome in patients undergoing solid organ transplantation (the ImmuneMo:SOT study): a prospective non-interventional observational trial

  Method Data output
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.