Flow cytometric patterns in blood from dogs with non-neoplastic and neoplastic hematologic diseases using double labeling for CD18 and CD45
| Autor: | M. Elena Gelain, V. Spagnolo, Fulvio Riondato, Stefano Comazzi, Paola Sartorelli, Roberta Guglielmino |
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| Rok vydání: | 2006 |
| Předmět: |
Pathology
medicine.medical_specialty medicine.drug_class Population Monoclonal antibody Cell morphology Dogs Antigen Internal medicine medicine Leukocytes Animals Dog Diseases education Settore VET/03 - Patologia Generale e Anatomia Patologica Veterinaria Acute leukemia education.field_of_study Hematology Leukemia General Veterinary biology Staining and Labeling medicine.disease Flow Cytometry Hematologic Diseases CD18 Antigens Immunology biology.protein Leukocyte Common Antigens Antibody |
| Popis: | Background In dogs, flow cytometry is used in the phenotyping of immunologic cells and in the diagnosis of hemic neoplasia. However, the paucity of specific antibodies for myeloid cells and B lymphocytes and of labeled antibodies for multicolor techniques limits the ability to detect all leukocyte subpopulations. This is especially true for neoplastic and precursor cells. CD18 and CD45 are expressed on all leukocytes and are involved in cell activation, and together could be useful in helping determine cell lineage. Objectives The purpose of this study was to double label canine blood for CD18 and CD45 and to use the differential expression of antigens to identify leukocyte populations in dogs with non-neoplastic and neoplastic hematologic diseases. Methods A template was developed using blood samples from 10 clinically healthy dogs and a back-gating technique. Differential leukocyte counts obtained with the template were compared with those obtained by manual and automated methods on blood samples from 17 additional healthy dogs. Blood samples obtained from 9 dogs with non-neoplastic (reactive) hematologic diseases and 27 dogs with hemic neoplasia were double stained for CD18 and CD45 using mouse anticanine CD18 monoclonal antibody (mAb) plus phycoerythrin-conjugated rat anticanine CD45 mAb and fluorescein isothiocyanate-conjugated rabbit antimouse IgG. Hemic neoplasms were diagnosed by cell morphology, and immunophenotypic and cytochemical markers. Results With the double label, neutrophils, eosinophils, monocytes, and T- and B-lymphocytes were identified. In reactive disorders, a population of activated neutrophils with high CD45 and CD18 expression was detected. In hemic neoplasia, cell lineage was easily determined, even in acute leukemia. Conclusions Double labeling for CD18/CD45 may be useful as a screening method to evaluate hematologic diseases and help determine cell lineage, and to aid in the selection of a panel of antibodies that would be useful for further analysis. |
| Databáze: | OpenAIRE |
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