Geuna, M.

Guglielmino, R

Miniscalco, Barbara

Morello, Emanuela Maria

Tarducci, Alberto

English

Institutional Research Information System University of Turin

Veterinary Research Communications, 27 Suppl. 1 (2003) 407–409© 2003 Kluwer Academic Publishers. Printed in the NetherlandsClinical Usefulness of Peripheral Blood Lymphocyte Subsets inCanine LymphomaB. Miniscalco1*, R. Guglielmino1, E. Morello1, A. Tarducci1 and M. Geuna21Department of Animal Pathology, University of T urin, Grugliasco (T urin);2L aboratory of T umor Immunology, I.R.C.C., Candiolo, Italy*Correspondence: Dipartimento di Patologia Animale, Facolta` di MedicinaVeterinaria, V ia L . da V inci, 44, 10095 Grugliasco (TO), ItalyE-mail: barbara.miniscalco@unito.itKeywords: blood, dog, flow cytometry, immunophenotype, lymphomaINTRODUCTIONThe classification of canine lymphoid malignancies into groups based on cell of origin(B- or T-cell types) is possible with immunophenotypic analysis. A high frequency ofB-cell lymphomas has been described; about 30% of lymphomas are of T-cell origin(Teske et al., 1994a; Fournel-Fleury et al., 1997; Grindem et al., 1998). In humanmedicine the major diagnostic role of blood lymphocyte subset typing is the screeningfor non-Hodgkin’s lymphomas with peripheral blood involvement and theirdifferentation from reactive lymphoproliferative conditions (Thalhammer-Scherreret al., 2000). The aim of this project is to evaluate the diagnostic efficacy of bloodimmunophenotyping in dogs with lymphoma in order to detect the blood involve-ment and the prognostic benefits.MATERIALS ANDMETHODSForty-eight dogs, 32 males and 16 females, ranging from 3 to 14 years old, of varyingbreeds, were examined. The diagnosis of lymphoma was performed on cytologicalsmears. The immunophenotype was detected from lymph node cellular suspensionsor cavity fluids using flow cytometry (FACScan Becton Dickinson): 36 cases wereCD21+ B-cell, 7 were CD4+ T-cell, 3 were CD8+ T-cell and 2 were CD4−CD8−T-cell lymphomas. A complete blood count and a phenotypic analysis of bloodlymphocyte subpopulations were performed. The following monoclonal antibodies,specific for canine leukocyte antigens, were used: CD21 (CA2.1D6), IgG F(ab∞)2 andIgM, CD5 (DH3B), CD3 (CA17.2A12), CD4 (CA13.1E4), CD8 (CA9.JD3), CD45RA(CA4.1D3), CD49d (CA4.5B3) and CD45 (CA12.10C12). Except for CD5 (VMRD,Pulmann, Washington, USA) and IgG, IgM (Jackson Immunoresearch, West Grove,PA, USA), all monoclonal antibodies were kindly provided by Dr P.F. Moore (U.C.407408Davies, CA, USA). The results were compared to reference values of a control goupof 50 healthy dogs (Guglielmino et al., 1995).RESULTSIn agreement with Jain (1993), the complete blood count showed a mild to modestanaemia in 46% of the dogs, thrombocytopenia in 44% and mild (18,300/ml ) toextreme (342,000/ml ) leukocytosis in 33%. Only 19% of cases exhibited lymphocyto-sis; lymphoblasts were present for 12 dogs (25%), and 15%were without lymphocyto-sis. In 10% of cases leukopenia was observed and lymphopenia was observed in 25%of cases. Five out of 36 (14%) B-cell lymphomas showed CD21+ B-cell lymphocytosis(ranging from 5520 to 115,670/ml ); four cases showed a moderate increase of B lym-phocytes without lymphocytosis. A CD8+ T-cell increase was observed in seven B-celllymphomas. This is also noted for human patients with neoplastic disorders, wherean increase in the so-called ‘anti-tumour’ cytolytic CD8+ T lymphocytes occurs(Quaglino et al., 1993). Two out of seven CD4+ T-cell lymphomas showed lympho-cytosis (17,680 and 45,050/ml ) with an increase in CD4+ T lymphocytes. Five dogs(71%) exhibited an increased CD4/CD8 ratio: in three cases this was due to anabsolute increase in CD4+ T cells. One dog showed an abnormal immunophenotype(TCD4+CD45− ) both in the lymph node and in the peripheral blood. In two out ofthe three dogs with CD8+ T-cell lymphoma a mild increase in the absolute count ofCD8+ T lymphocytes, without lymphocytosis, was noted; one of these showed thesame abnormal immunophenotype as the lymph node (TCD8+CD45− ). Extremelymphocytosis (12,690 and 338,580/ml ), due to neoplastic cells, was observed for twodogs with CD4−CD8− T-cell lymphoma.DISCUSSIONIn dogs with lymphoma the total and differential leukocyte counts and the presenceof atypical lymphocytes were very variable: indeed blood involvement was not alwaysdetectable on a simple complete blood count. The detection of neoplastic cells (5T-cell and 4 B-cell lymphomas) with the same immunophenotype as the origintumours, also without lymphocytosis and cells with atypical morphology, was pos-sible using flow cytometric analysis of peripheral blood. The immunophenotype ofthe peripheral blood was able to provide more precise information about systemicinvolvement. In 75% (9/12) of T-cell lymphomas and in 25% (9/36) of B-cell lympho-mas a systemic involvement was present, confirming a poorer prognosis for T-celllymphomas as compared with B-cell lymphomas (Teske et al., 1994b). Simultaneousimmunophenotyping of blood, lymph node and bone marrow from dogs with lym-phoma could represent an important diagnostic and prognostic tool. Our studysupports the potential use of blood immunophenotyping as a sensitive tool in the409detection of leukaemic cells, facilitation of tumour staging and monitoring ofremission or relapse.ACKNOWLEDGEMENTSThis research was supported by a grant from the Italian MURST.REFERENCESFournel-Fleury, C., Magnol, J.P., Bricaire, P., Marchal, T., Chabanne, L., Delverdier, A., Bryon, P.A. andFelman, P., 1997. Cytohistological and immunological classification of canine malignant lymphomas.Comparison with human non-Hodgkin’s lymphomas. Journal of Comparative Pathology, 117, 35–59Grindem, C.B., Page, R.L., Ammerman, B.E. and Breitschwerdt, E.B., 1998. Immunophenotipic comparisonof blood and lymph node from dogs with lymphoma. Veterinary Clinical Pathology, 27(1), 16–20Guglielmino, R., Geuna, M., Tarducci, A., D’Angelo, A., Ru, G. and Bruatto, G., 1995. Analisi citofluori-metrica delle sottopopolazioni linfocitarie nel sangue di 50 cani sani. Atti della Societa` Italiana delleScienze Veterinarie, 49, 811–812Jain, N.C., 1993. Essentials of Veterinary Hematology, (Lea & Febiger, Philadelphia), 334–335Quaglino, D., De Pasquale, A. and Ginaldi, L., 1993. Hematological changes in malignant disease. EuropeanJournal of L aboratory Medicine, 1, 23–33Teske, E., Wisman, P., Moore, P.F. and Van Heerde, P., 1994a. Histological classification and immunophe-notyping of canine non-Hodgkin’s lymphomas. Unesxpected high frequency of T-cell lymphomas withB-cell morphology. Experimental Hematology, 22, 1179–1187Teske, E., Van Heerde, P., Rutteman, G.R., Kurzman, I.D., Moore, P.F. and MacEwen, E.G., 1994b.Prognostic factors for treatment of malignant lymphoma in dogs. Journal of American VeterinaryMedical Association, 205, 1722–1728Thalhammer-Scherrer, R., Veitl, M., Exner, M., Schneider, B., Geissler, K., Simonitsch, I. andSchwarzinger, I., 2000. Role of immunological lymphocyte subset typing as a screening method forlymphoid malignancies in daily routine practice. Cytometry, 42, 5–10

Clinical usefulness of peripheral blood lymphocyte subsets in canine lymphoma

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Clinical usefulness of peripheral blood lymphocyte subsets in canine lymphoma

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