The Use of Cytochemistry, Immunophenotyping, Flow Cytometry, and in Vitro Differentiation to Determine the Ontogeny of a Canine Monoblastic Leukemia

Jaime F. Modiano, Roger Smith, John Wojcieszyn, Jennifer S. Thomas, Betty A. Rosenbaum, Carrie Ball, Elaine A. Nicholds, Margaret A. Anthony, Claudia L. Barton

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

We evaluated the utility of cytochemistry, immunophenotyping, flow cytometry, and in vitro culture with forced differentiation of leukemic cells as diagnostic aids to identify the malignant cell ontogeny in a dog with leukemia. A tentative diagnosis of monoblastic leukemia was established by microscopic examination of Romanowsky-stained blood smears and bone marrow aspirate smears. This diagnosis also was supported by the light scatter signature that identified the blast cells as large, non-granular monocytic cells using a CellDyn 3500 automated hematology analyzer; as well as by the detection of N-butyrate esterase and the lack of chloroacetate esterase or leukocyte peroxidase by cytochemical staining. Subsequently, leukemic cells were isolated from the dog's peripheral blood and placed into tissue culture or cryopreserved. The leukemic cells grew in suspension cultures and proliferated spontaneously for up to 4 days. By day 7, proliferation was negligible. Upon culture with conditioned supernatant using mitogen-stimulated human T cells as a source of cytokines, an increased proportion of cells entered S phase by day 2 of culture; however, proliferation declined markedly by day 4, at which time the cells had apparently differentiated to adherent, vacuolated macrophages. The cytokine-stimulated leukemic cells were positive for the monocyte/macrophage specific markers alpha-1-antitrypsin, alpha-1-antichymotrypsin, lysozyme, CD14, MHC class II, and calprotectin, an antigen found in differentiated macrophages and granulocytes. Despite the strong tendency of the leukemic cells towards monocytic differentiation, our results suggested that they retained some features of a myelomonocytic precursor. These data show that cytochemistry, immunophenotyping, flow cytometry, and in vitro differentiation of canine leukemia cells are useful tools for confirming the lineage of malignant hematopoietic cells.

Original languageEnglish (US)
Pages (from-to)40-49
Number of pages10
JournalVeterinary Clinical Pathology
Volume27
Issue number2
DOIs
StatePublished - Jan 1 1998

Fingerprint

cytochemistry
Immunophenotyping
Histocytochemistry
leukemia
flow cytometry
ontogeny
Canidae
Flow Cytometry
Leukemia
dogs
cells
macrophages
Macrophages
esterases
cytokines
In Vitro Techniques
alpha 1-Antichymotrypsin
Leukocyte L1 Antigen Complex
Dogs
Cytokines

Keywords

  • Automated hematology analyzer
  • Cytochemistry
  • Flow cytometry
  • Immunophenotyping
  • Leukemia
  • Monocyte differentiation

Cite this

The Use of Cytochemistry, Immunophenotyping, Flow Cytometry, and in Vitro Differentiation to Determine the Ontogeny of a Canine Monoblastic Leukemia. / Modiano, Jaime F.; Smith, Roger; Wojcieszyn, John; Thomas, Jennifer S.; Rosenbaum, Betty A.; Ball, Carrie; Nicholds, Elaine A.; Anthony, Margaret A.; Barton, Claudia L.

In: Veterinary Clinical Pathology, Vol. 27, No. 2, 01.01.1998, p. 40-49.

Research output: Contribution to journalArticle

Modiano, Jaime F. ; Smith, Roger ; Wojcieszyn, John ; Thomas, Jennifer S. ; Rosenbaum, Betty A. ; Ball, Carrie ; Nicholds, Elaine A. ; Anthony, Margaret A. ; Barton, Claudia L. / The Use of Cytochemistry, Immunophenotyping, Flow Cytometry, and in Vitro Differentiation to Determine the Ontogeny of a Canine Monoblastic Leukemia. In: Veterinary Clinical Pathology. 1998 ; Vol. 27, No. 2. pp. 40-49.
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