König, Maya L.; Lettry, Sophie C.; Marti, Eliane; Mirkovitch, Jelena; Wyder, Marianne; Giger, Urs; Schuller, Simone (2018). Validation of a human angiopoietin-2 enzyme linked immunosorbent assay kit for measurement of canine angiopoietin-2 concentrations in plasma and supernatant of primary canine aortic endothelial cell cultures. American journal of veterinary research, 79(8), pp. 803-810. American Veterinary Medical Association 10.2460/ajvr.79.8.803
Full text not available from this repository. (Request a copy)Objective – To assess two human ELISA kits for measurement of angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) in canine plasma and to evaluate the effect of sepsis on plasma Ang-2 concentrations. To isolate and culture primary canine aortic endothelial cells (pCAEC) and study the effect of TNF-α stimulation on Ang-2 release from pCAEC.
Animals –Ten healthy dogs and ten dogs with bacterial sepsis.
Procedures – Human Ang-1 and Ang-2 ELISA kits were assessed using recombinant canine Ang-1 and Ang-2 and canine plasma. The Ang-1 ELISA did not detect canine Ang-1. The Ang-2 ELISA detected recombinant canine Ang-2 and was further validated using plasma of healthy and septic dogs, and supernatants of pCAEC cultures. Dilution linearity, parallelism, accuracy, precision, and reproducibility of the assay were determined. Associations between plasma Ang-2 and C-reactive protein (CRP) concentrations were examined.
Results – The human Ang-2 ELISA had excellent dilutional linearity, parallelism, accuracy, precision, and reproducibility for measurement of Ang-2 in canine plasma and endothelial cell supernatants. Plasma Ang-2 was significantly higher in septic (median 25.5; IQR, 19.9–69.9 ng/mL) than in healthy dogs (median 6.7; IQR, 5.9–9.8 ng/mL; p<0.002) and positively correlated with plasma CRP concentrations (r2=0.6; p<0.001). Angiopoietin-2 markedly increased in supernatants of pCAEC after TNF-α stimulation.
Conclusions and Clinical Relevance – The tested human Ang-2 ELISA is useful for measurement s of canine Ang-2 concentrations accurately. Plasma Ang-2 concentrations are increased in septic dogs. Angiopoietin-2 is released from pCAEC after TNF-α stimulation. This study provides new tools to study Ang-2 as marker of endothelial cell activation and inflammation in dogs.
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