Measuring cytokine levels in blood: Importance of anticoagulants, processing, and storage conditions

摘要: The stability and recovery of six human recombinant cytokines (tumour necrosis factor (TNF), interferon-alpha (IFN-alpha), IFN-gamma, interleukin-1 alpha (IL-1 alpha), IL-1 beta, IL-6) from whole blood was investigated with a view to optimizing collection storage procedures prior performing immunoassays. Blood healthy volunteers subjected various processing procedures. samples were treated either: ethylenediamine tetraacetic acid (EDTA) (1.5 mg/ml blood) (E); EDTA/Trasylol mg 1000 KIU/ml (ET); heparin (30 IU/ml) (H) or allowed clot (serum). bloods spiked individual cytokines, split into aliquots kept at 4 degrees C RT. In the first instance (n = 5 per cytokine) processed using sterile non-pyrogenic materials At regular time intervals, cold spun, separated, flash frozen assayed for appropriate cytokine RIA/IRMA methods. further study, timed separation repeated normal commercially available third 3 under conditions, aliquoted within half hour collection. These then cycles freeze thawing RT before assaying. general, in improved by and/or rapid separation. There no significant difference between handled sterile, conditions those collected described this paper did not induce any unspiked blood. Overall, EDTA-treated performed most consistently. addition trasylol significantly affect results. Most appeared unaffected up three thaw cycles. varied least stable as follows; TNF-alpha less than IL-6 IFN-gamma IFN-alpha beta. heparinized plasma considerably higher other serum samples. consistently lower amounts recovered (anticoagulant treated).(ABSTRACT TRUNCATED AT 400 WORDS)