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Flow cytometric immunophenotype of canine lymph node aspirates
D Gibson1, I Aubert, J P Woods
1Department of Pathobiology, University of Guelph, Guelph, ON, Canada.
Insights
Flow cytometry on canine lymph node aspirates is feasible for diagnosing lymphoma. This method provides reliable immunophenotypic results, even after overnight storage, aiding in canine cancer diagnosis.
Area of Science:
- Veterinary immunology
- Canine pathology
- Flow cytometry applications
Background:
- Advances in reagent availability enhance understanding of canine lymphocyte biology and pathology.
- Flow cytometry is a common method for assessing circulating lymphocytes, but lymph node aspirates are also suitable.
- Needle aspirates offer a minimally invasive approach for lymph node analysis.
Purpose of the Study:
- To determine the feasibility of immunophenotyping canine lymph node aspirates using flow cytometry.
- To establish reference intervals for lymphocyte subsets in normal dogs.
- To assess the diagnostic utility of this technique for canine lymphoma.
Main Methods:
- Collected lymph node aspirates from 32 normal dogs and 6 dogs with lymphoma.
- Performed microscopic examination of stained smears for cell morphology and inflammation.
- Analyzed cell surface antigen expression via flow cytometry on at least 10,000 events per sample.
- Assessed sample stability after overnight storage at 4°C.
Main Results:
- Adequate samples were obtained from 30 of 32 clinically normal dogs.
- Immunophenotyping results were consistent after 24 hours of sample storage.
- Established reference intervals for lymphocyte subsets (CD3+, CD4+, CD8+, CD21+) in normal dogs.
- Lymph node aspirates from dogs with lymphoma were readily classified by flow cytometry.
Conclusions:
- Lymph node aspiration is a feasible method for comprehensive flow cytometric analysis in dogs.
- This technique is applicable for the immunophenotyping and diagnosis of canine lymphoma.
- The findings support the use of lymph node aspirates for routine diagnostic workups in veterinary oncology.
Abstract:
Increasing availability of reagents able to distinguish subtypes of lymphocytes and other leukocytes has enabled greater understanding of lymphocyte biology and pathology in the dog. Lymphocytes in circulation most commonly are subjected to immunophenotypic assessment by flow cytometry, but needle aspirates of lymph nodes can be similarly suitable for immunophenotypic examination. In this investigation, the feasibility of immunophenotyping samples obtained by needle aspiration of lymph nodes from 32 dogs with no physical abnormalities and 6 dogs with lymphoma was determined. In addition, samples from 6 dogs were stored overnight at 4 degrees C and reanalyzed 24 hours later. For each sample, stained smear preparations were examined microscopically for lymphocyte morphology, neoplasia, and the presence of inflammatory cells. Expression of antigens on a corresponding sample of aspirated cells was determined by flow cytometric detection of antibody binding on a minimum of 10,000 events. The distribution of data was determined with Anderson-Darling tests, and reference intervals incorporating the central 95% of values were established. Adequate samples were obtained from 30 of 32 clinically normal dogs. Immunophenotypic results after 24 hours of storage were consistent with those obtained immediately after sampling. Reference intervals for lymphocyte subsets from normal dog lymph nodes were similar to the proportions of CD3+, CD4+, CD8+, and CD21+ lymphocytes found in blood. Aspirates of enlarged lymph nodes from dogs with lymphoma were readily classified by this technique. Aspiration of lymph nodes from dogs for comprehensive analysis by flow cytometry is feasible and applicable to immunophenotyping of lymphoma.
