Immunophenotyping pattern characterization in canine blood: towards a clinical application

León, Leticia G.; Lopez, Fatima Cruz; Fermín, M. Luisa; Mejías, Guillermo; Kremmer, Elisabeth; Tejero, Concepción

doi:10.2754/avb201685020139

Acta Veterinaria > Archive > 2016, Vol. 85 > Issue 2 > Immunophenotyping pattern…

Acta Vet. Brno 2016, 85: 139-145

https://doi.org/10.2754/avb201685020139

Immunophenotyping pattern characterization in canine blood: towards a clinical application

Leticia G. León¹, Fatima Cruz Lopez², M. Luisa Fermín³, Guillermo Mejías⁴, Elisabeth Kremmer⁵, Concepción Tejero⁴

¹Centre for Biomedical Research of the Canary Islands (CIBICAN) ULL, Institute of Biomedical Technologies, Tenerife, Spain
²Complutense University of Madrid, Veterinary Hospital Clinic, Laboratory of Clinical Pathology, Madrid, Spain
³Complutense University of Madrid, Veterinary Faculty, Department of Animal Surgery and Medicine, Madrid, Spain
⁴Complutense University of Madrid, Veterinary Faculty, Department of Biochemistry annd Molecular Biology, Madrid, Spain
⁵Institute for Molecular Immunology, Helmholtz Zentrum München, Munich, Germany

Received July 13, 2015
Accepted May 2, 2016

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.