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Published on: January 7, 2019
Aberrant phenotypes and quantitative antigen expression in different subtypes of canine lymphoma by flow cytometry
M E Gelain1, M Mazzilli, F Riondato
1Department of Veterinary Pathology, Hygiene and Health, University of Milan, via Celoria 10, Milan 20133, Italy. mariaelena.gelain@unimi.it
Insights
Flow cytometry provides objective data on lymphoma cell morphology and immunophenotype, aiding diagnosis. This technique enhances conventional cytology by identifying aberrant antigen expression in canine lymphoma cases.
Area of Science:
- Veterinary Pathology
- Immunology
- Cytopathology
Background:
- Fine needle aspiration (FNA) is a common method for obtaining lymphoma samples.
- Conventional cytology provides morphological assessment but can lack objective, standardized data.
- Flow cytometry offers a potential method to enhance diagnostic accuracy by analyzing cellular characteristics.
Purpose of the Study:
- To evaluate flow cytometry's utility in analyzing canine lymphoma FNA samples.
- To determine if flow cytometry adds objective information on cellularity and morphology compared to conventional cytology.
- To assess typical immunophenotypes and aberrant antigen expression in lymphoma subtypes.
Main Methods:
- Analysis of 50 canine lymphoma FNA lymph node samples.
- Flow cytometric immunophenotyping to identify B-cell, T/NK-cell origins, and acute lymphoblastic leukemia.
- Evaluation of leukocyte marker expression for antigen up/down-regulation.
Main Results:
- 31 B-cell, 16 T/NK-cell lymphomas, and 3 acute lymphoblastic leukemia cases identified.
- Aberrant phenotypes observed in 64% of lymphoma cases (58% B-cell, 75% T-cell).
- Diminished CD79a expression in 45% of B-cell lymphomas; CD45 under-expression in 25% of T-cell lymphomas.
Conclusions:
- Flow cytometry is a valuable tool for confirming canine lymphoma diagnoses.
- The technique provides objective data on neoplastic cell morphology and immunophenotype.
- Identification of aberrant antigen expression aids in lymphoma subtyping and diagnosis.
Abstract:
Flow cytometry may be a useful tool to analyze lymphoma samples that are obtained from fine needle aspirations (FNA). This study aimed to determine if flow cytometric analysis add more objective and standardized information on the cellularity and morphology of lymphoma cells to conventional cytology. The typical immunophenotype of different lymphoma subtypes was assessed and leukocyte marker expression was evaluated to determine which antigens were more frequently over- or under-expressed in these lymphoma subtypes. Fifty FNA lymph node samples were evaluated from canine lymphomas. Thirty-one samples were identified to be of B-cell origin, sixteen were identified to be of T/NK-cell origin and three cases were classified as acute lymphoblastic leukaemia with lymph nodes involvement. The most common B-cell lymphoma subtypes were centroblastic lymphomas, whereas three cases were atypical and classified as B-large cell pleomorphic lymphomas. Among the T/NK lymphomas, small clear cells, large and small pleomorphic mixed cells, large granular lymphocytic cells and small pleomorphic cells were identified. Aberrant phenotypes and/or antigen under/over regulation was identified in thirty out of forty-seven lymphoma cases (64%; 18/31 B-cell=58% and 12/16 T-cell=75%). In B-cell lymphomas the most frequent finding was the diminished expression of CD79a (45%). CD34 expression was also observed in four cases (13%). Among T-cell lymphomas the prevalent unusual phenotype was the under-expression or absence of CD45 (25%). These findings reveal flow cytometry may be useful in confirming the diagnosis of lymphoma, as the technique allows one to add useful information about morphology of the neoplastic cells and identify antigenic markers and aberrant phenotypes.
