Angiopoietin-2 is a novel marker of endothelial dysfunction and vascular leakage. The aim of this study was to validate a human Enzyme-linked Immunosorbent Assay (ELISA) for measurement of angiopoietin-2 (Ang-2) in canine plasma and to evaluate the effect of SIRS/sepsis on plasma Ang-2 concentrations in dogs. Ethical approval for collection of blood samples from healthy dogs and dogs with SIRS/sepsis was obtained (26425 BE38/15). The assay was validated using canine plasma samples, canine recombinant Ang-2 and supernatants from quiescent and stimulated canine aortic endothelial cells. Dilution linearity and parallelism, accuracy, precision, and reproducibility of the assay were determined. The association of age, sex, weight, and sepsis with Ang-2 plasma concentration were analysed.

The ELISA had excellent dilution linearity, parallelism, accuracy, precision, and reproducibility for measurement of Ang-2 concentrations in canine plasma and endothelial cell supernatants. Median plasma Ang-2 concentration was significantly higher in septic dogs (25.5; IQR, 19.9–69.9 ng/ml) than in healthy dogs (6.7; IQR, 5.9–9.8 ng/ml). Ang-2 concentration in supernatant of canine macrovascular endothelial cells was significantly higher after stimulation with tumour necrosis factor a (TNF-a) compared to unstimulated cells.

The tested human ELISA is able to measure canine Ang-2. Data suggests that Ang-2 is increased in plasma of dogs with sepsis and released from endothelial cells upon stimulation with TNF-a, similar to what has been described in human medicine. This data provides the basis for future studies using Ang-2 as a marker of endothelial cell activation in dogs.

Disclosures

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