Acta Vet. Brno 2016, 85: 139-145

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


1. Cobbold S, Metcalfe S 1994: Monoclonal antibodies that define canine homologues of human CD antigens: summary of the First International Canine Leukocyte Antigen Workshop (CLAW). Tissue Antigens 43: 137-154 <>
2. De Paoli P, Reitano M, Battistin S, Castiglia C, Santini G 1984: Enumeration of human lymphocyte subsets by monoclonal antibodies and flow cytometry: a comparative study using whole blood or mononuclear cells separated by density gradient centrifugation. J Immunol Methods 72: 349-353 <>
3. Faldyna M, Levá L, Knötigová P, Toman M 2001: Lymphocyte subsets in peripheral blood of dogs-a flow cytometric study. Vet Immunol Immunopathol 82: 23-37 <>
4. Hardy MY, Vari F, Rossetti T, Hart DN, Prue RL 2013: A flow cytometry based assay for the enumeration of regulatory T cells in whole blood. J Immunol Methods 390: 121-126 <>
5. Herzenberg LA, Tung J, Moore WA, Herzenberg LA, Parks DR 2006: Interpreting flow cytometry data: a guide for the perplexed. Nat Immunol 7: 681-685 <>
6. Kolb HJ, Günther W, Schumm M, Holler E, Wilmanns W, Thierfelder S 1997: Adoptive immunotherapy in canine chimeras. Transplantation 63: 430-436 <>
7. Krause DS, Fackler MJ, Civin CI, May WS 1996: CD34: structure, biology, and clinical utility. Blood 87: 1-13
8. Mansour I, Bourin P, Rouger P, Doinel C 1990: A rapid technique for lymphocyte preparation prior to two-color immunofluorescence analysis of lymphocyte subsets using flow cytometry. Comparison with density gradient separation. J Immunol Methods 127: 61-70 <>
9. McSweeney PA, Rouleau KA, Wallace PM, Bruno B, Andrews RG, Krizanac-Bengez L, Sandmaier BM, Storb R, Wayner E, Nash RA 1998: Characterization of monoclonal antibodies that recognize canine CD34. Blood 91: 1977-1986
10. Nicholson JK, Jones BM, Cross GD, McDougal JS 1984: Comparison of T and B cell analyses on fresh and aged blood. J Immunol Methods 73: 29-40 <>
11. Ostronoff LK, Kremmer E, Ferm’n ML, Frag’o C, Mysliwietz J, Kolb HJ, Tejero C 2008: Canine stem cell factor augments expression of matrix metalloproteinase-9 by CD34 cells. Cytotherapy 10: 193-202 <>
12. Owens MA, Vall HG, Hurley AA, Wormsley SB 2000: Validation and quality control of immunophenotyping in clinical flow cytometry. J Immunol Methods 243: 33-50 <>
13. Ponzio AD, Self SE, Runey DH, Burdash NM, La Via MF 1984: Are lymphocyte subset determinations affected by storage conditions? Diagn Immunol 2: 188-190
14. Prince HE, Arens L 1986: Effect of storage on lymphocyte surface markers in whole blood units. Transplantation 41: 235-238 <>
15. Schwonzen M, Diehl V, Dellanna M, Staib P 2007: Immunophenotyping of surface antigens in acute myeloid leukemia by flow cytometry after red blood cell lysis. Leuk Res 31: 113-116 <>
16. Shield CF 3rd, Marlett P, Smith A, Gunter L, Goldstein G 1983: Stability of human lymphocyte differentiation antigens when stored at room temperature. J Immunol Methods 62: 347-352 <>
17. Van Dongen JJ, Langerak AW, Brźggemann M, Evans PA, Hummel M, Lavender FL, et al 2003: Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED-2 Concerted Action BMH4-CT98-3936. Leukemia 17: 2257-2317 <>
18. Watabe A, Hanazono K, Komatsu T, Fu DR, Endo Y, Kadosawa T 2012: Peripheral lymphocyte subsets as a prognostic indicator of mortality and morbidity in healthy dogs. J Vet Med Sci 74: 937-943 <>
19. Weiss DJ 2002: Application of flow cytometric techniques to veterinary clinical hematology. Vet Clin Pathol 31: 72-82 <>
20. Williams DL 1997: Studies of canine leucocyte antigens: a significant advance in canine immunology. Vet J 153: 31-39 <>
21. Williams MJ, Avery AC, Lana SE, Hillers KR, Bachand AM, Avery PR 2008: Canine lymphoproliferative disease characterized by lymphocytosis: immunophenotypic markers of prognosis. J Vet Intern Med 22: 596-601 <>
22. Zeigler BM, Boyle-Holmes Y, Falzone D, Farmer J 2013: Validation of an eight parameter immunophenotyping panel in adult canines for assessment of immunotoxicity. Vet Immunol Immunopathol 154: 75-81 <>
front cover
  • ISSN 0001-7213 (printed)
  • ISSN 1801-7576 (electronic)

Current issue

Indexed in DOAJ