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Immunophenotyping of Orthotopic Homograft (Syngeneic) of Murine Primary KPC Pancreatic Ductal Adenocarcinoma by Flow Cytometry
Published on: October 9, 2018
A comparative study of frozen-section immunoperoxidase and flow cytometry for immunophenotypic analysis of lymph node
K W Biesemier1, G A Dent, K B Pryzwansky
1Department of Pathology, University of North Carolina-Chapel Hill 27514, USA.
Insights
Flow cytometry and frozen-section immunoperoxidase show high concordance for lymphoma immunophenotyping. Flow cytometry excels at detecting light-chain restriction, crucial for accurate lymphoma diagnosis.
Area of Science:
- Hematology
- Immunology
- Pathology
Background:
- Accurate immunophenotyping of lymph node biopsies is critical for lymphoma diagnosis.
- Flow cytometry and frozen-section immunoperoxidase are common diagnostic methods.
- Comparing these techniques can optimize diagnostic workflows.
Purpose of the Study:
- To compare the concordance of flow cytometry and frozen-section immunoperoxidase for lymph node immunophenotyping.
- To evaluate the diagnostic performance of each method for detecting light-chain restriction.
- To assess the reliability of fine-needle aspiration for flow cytometry sample acquisition.
Main Methods:
- Immunophenotyping was performed on 21 lymph node biopsy specimens using both flow cytometry and frozen-section immunoperoxidase.
- Flow cytometry samples were obtained via fine-needle aspiration.
- Concordance and diagnostic values (sensitivity, specificity, predictive values) were calculated for routine markers and light-chain restriction.
Main Results:
- Overall concordance ranged from 76% to 100%, with B-cell markers showing lower concordance than T-cell markers.
- Flow cytometry demonstrated significantly higher sensitivity (90% vs. 30%) and negative predictive value (100% vs. 63%) for light-chain restriction.
- Both methods accurately identified characteristic immunophenotypes for various lymphoma subtypes.
Conclusions:
- Flow cytometry and frozen-section immunoperoxidase are largely concordant methods for lymph node immunophenotyping.
- Flow cytometry is superior for demonstrating light-chain restriction, aiding in lymphoma diagnosis.
- Ex vivo fine-needle aspiration is a reliable method for obtaining lymph node cell suspensions for flow cytometry.
Abstract:
Immunophenotyping by flow cytometry and frozen-section immunoperoxidase was compared on 21 consecutive lymph node biopsy specimens, of which a diagnosis of lymphoma was made for 11 specimens. Samples for flow cytometry were obtained by a fine-needle aspiration technique. Concordance between frozen-section immunoperoxidase and flow cytometry for all routine markers on all specimens ranged from 76 to 100%. In general, B-cell markers showed poorer concordance than T-cell markers, with kappa and lambda light chains having the poorest concordance, at 76% each. Flow cytometry was significantly more sensitive (90 versus 30%; P < 0.006) and had a significantly higher negative predictive value (100 versus 63%; P < 0.006) than frozen-section immunoperoxidase for demonstrating light-chain restriction. There was no significant difference in the specificities (100 versus 91%) or positive predictive values (100% each) between the two methods. Both methods demonstrated characteristic immunophenotypes for intermediate cell lymphomas, small lymphocytic lymphomas, and T-cell lymphoblastic lymphomas. Frozen-section immunoperoxidase and flow cytometry appear to be significantly concordant methods for immunophenotypic analysis of lymph node biopsies. Light-chain restriction is more readily demonstrated by flow cytometry than frozen-section immunoperoxidase. We believe that ex vivo fine-needle aspiration is a simple and reliable method of obtaining cell suspensions of lymph nodes for flow cytometry.
