Immunophenotyping pattern characterization in canine blood: towards a clinical application

Abstract

Immunophenotyping is a widely used method for a precise diagnosis and classification of haematopoietic neoplasia in human beings and also in dogs. The gold standard for cell preparation is density gradient centrifugation of mononuclear cells. Alternatively, another way to separate human leukocytes is carrying out whole blood lysis. The aim of this study was to validate whole blood lysis as an alternative method in clinical veterinary procedures using an immunophenotype panel of leukocytes designed by our group. Flow cytometry study of adult canine leukocytes subset groups, using whole blood lysis or mononuclear cells tested against an array of canine leukocyte antibodies were done. Besides differential white blood cell counts were done. Also immunophenotyping studies in whole blood samples stored at 4 °C for 48 h were performed. The Coefficient Variation values were less than 20%, for most of the comparison. Consistent results were observed in phenotyping canine peripheral blood leukocytes. Stability results indicated that whole blood samples might be stored for 48 h without a significant difference in the data compared to samples processed immediately after blood collection. This study shows that whole blood lysis represents an efficient and quick alternative for canine leukocyte preparation. In addition, samples can be analysed immediately or stored for 48 h without a significant difference between them. This is relevant for veterinary medicine considering the lack of facilities in many laboratories to process samples. Flow cytometry, peripheral blood, leukocyte subsets, long-term stability