Location, location, location: cytokine concentrations are dependent on blood sampling site.

objectiveConsiderable breakthroughs in the field of sepsis have been made using animal models. Sepsis exhibits a wide array of derangements that may be evaluated in the blood, including the release of proinflammatory and anti-inflammatory cytokines. The Shock journal adheres to the ARRIVE guidelines regarding reporting in vivo results to allow reproducibility of data findings. It is generally assumed that blood cytokine concentrations collected from typical sampling sites will be similar, but there are no data validating that this is true. The main purpose of the present study was to determine if the location of blood sampling results in cytokine concentration differences following inflammatory insults.

methodsTwo different models of acute inflammation were studied. Adult, female ICR (Institute of Cancer Research) mice were injected with Escherichia coli lipopolysaccharide (n = 28) or subjected to cecal ligation and puncture (n = 16). They were killed at early time points following these inflammatory challenges for the collection of blood from the facial vein, retro-orbital sinus, and heart. Additional samples were collected in EDTA and heparin. Plasma cytokines from the same mouse were collected from each sampling site and evaluated by enzyme-linked immunosorbent assay. Clinical chemical parameters including plasma blood urea nitrogen and total protein were also analyzed.

resultsRegardless of model, time of collection, or cytokine measured, cytokine values from heart blood were higher than facial vein values from the same mouse. Interleukin (IL-6) collected from the heart relative to the facial vein demonstrated elevated concentrations following injection of lipopolysaccharide. In a similar manner, higher concentrations of IL-6, macrophage inflammatory protein 2, IL-10, and IL-1 receptor antagonist were found in cardiac puncture samples compared with other sampling sites 24 h after sepsis induced by cecal ligation and puncture. Similar differences were not seen when comparing blood urea nitrogen and total protein values from the two different sites. Using plasma IL-6 collected from the heart would incorrectly stratify predicted-to-live mice into the predicted-to-die category. Therefore, a simple linear regression model was developed to correctly restratify mice to their predicted fate. These data demonstrate that proinflammatory and anti-inflammatory cytokine concentrations are dramatically elevated when drawn centrally from the heart compared with collection from peripheral locations such as the facial vein. It is critical for publications to document the sampling location when evaluating plasma cytokines and attempting to compare studies.