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Flow cytometric immunophenotyping of anaplastic large cell lymphoma
Tariq Muzzafar1, Eric X Wei, Pei Lin
1Department of Hematopathology, The University of Texas M. D. Anderson Cancer Center, Houston, TX 77030, USA.
Insights
Flow cytometry effectively immunophenotypes Anaplastic Large Cell Lymphoma (ALCL) cases, even when neoplastic cells are scarce or atypical. Cluster analysis aids in identifying these cells for accurate diagnosis.
Area of Science:
- Hematopathology
- Immunology
- Oncology
Background:
- Anaplastic Large Cell Lymphoma (ALCL) diagnosis traditionally relies on histology and immunohistochemistry.
- Fine-needle aspiration (FNA) is an increasingly utilized alternative to lymph node biopsy.
- Flow cytometry is a key technique for analyzing FNA specimens in ALCL diagnosis.
Purpose of the Study:
- To review flow cytometry findings in ALCL cases.
- To evaluate the diagnostic utility of flow cytometry for ALCL.
- To assess the immunophenotypic characteristics of ALCL using flow cytometry.
Main Methods:
- Flow cytometry with 3-color or 4-color antibody panels was used for ALCL assessment.
- Cluster analysis with Paint-A-Gate software reanalyzed flow cytometry data.
- Immunohistochemistry was employed to assess anaplastic lymphoma kinase (ALK) status.
Main Results:
- Nineteen of 23 ALCL cases showed identifiable neoplastic cells via flow cytometry (11 ALK+, 8 ALK-).
- ALCL cells were typically CD30 and CD45 positive, often appearing large and CD45 bright.
- Distinct T-cell antigen expression profiles were observed between ALK(+) and ALK(-) ALCL cases.
Conclusions:
- Flow cytometry is a viable method for immunophenotyping ALCL.
- Neoplastic cells in ALCL may be sparse and located outside the typical lymphocyte gate.
- Cluster analysis software facilitates flexible gating strategies for ALCL cell identification and characterization.
Context:
Anaplastic large cell lymphoma (ALCL) is usually diagnosed by histologic and immunohistochemical analysis. However, fine-needle aspiration is becoming a popular alternative to lymph node biopsy, and flow-cytometric immunophenotyping is often used to analyze fine-needle aspiration specimens.
Objective:
To review our experience using flow-cytometric immunophenotyping to assess cases of ALCL and to evaluate the diagnostic utility of this technique.
Design:
Each case of ALCL was assessed by flow cytometry with 3-color or 4-color antibody panels, and data were reanalyzed by cluster analysis using Paint-a-Gate for cases with retrievable flow cytometry data files. Anaplastic lymphoma kinase (ALK) was assessed by using immunohistochemistry.
Results:
Twenty-three ALCL cases were analyzed by flow cytometry. In 4 cases, neoplastic cells could not be identified. In the remaining 19 cases (11 ALK(+), 8 ALK(-)), all were positive for CD30 and CD45. Anaplastic large cell lymphoma cells were large and usually CD45 bright, with many or most cells falling in the region of monocytes on the CD45/side scatter plot. The frequencies of T-cell antigen expression in ALK(+) cases were CD2, 67%; CD7, 60%; CD3, 45%; CD4, 33%; CD5, 14%; and CD8, 14%. In ALK(-) cases, the frequencies of the T-cell antigen expression were CD2, 100%; CD3, 50%; CD4, 40%; CD7, 40%; CD5, 25%; and CD8, 20%.
Conclusions:
Flow cytometry can be used to immunophenotype ALCL cases. Neoplastic cells may be few, and they may fall outside of the lymphocyte gate. Cluster analysis using software like Paint-A-Gate is often helpful because it allows for flexible, sequential gating strategies to identify and characterize the neoplastic cells.
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