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Updated: May 23, 2026
Flow-sorting and Exome Sequencing of the Reed-Sternberg Cells of Classical Hodgkin Lymphoma
Published on: June 10, 2017
Strategies for immunophenotyping and purifying classical Hodgkin lymphoma cells from lymph nodes by flow cytometry
Jonathan R Fromm1, Brent L Wood
1Department of Laboratory Medicine, University of Washington, Seattle 98195, USA. jfromm@u.washington.edu
Insights
Flow cytometry can now diagnose classical Hodgkin lymphoma (CHL) by identifying neoplastic Hodgkin and Reed-Sternberg (HRS) cells. This technique offers high sensitivity and specificity, aiding in diagnosis and cell purification.
Area of Science:
- Hematology
- Immunology
- Cell Biology
Background:
- Classical Hodgkin lymphoma (CHL) diagnosis traditionally relies on paraffin-embedded tissue sections.
- Neoplastic Hodgkin and Reed-Sternberg (HRS) cells are the hallmark of CHL.
- Flow cytometry is a powerful tool for immunophenotyping hematopoietic neoplasms.
Purpose of the Study:
- To demonstrate the utility of flow cytometry for identifying and immunophenotyping neoplastic HRS cells in CHL.
- To establish protocols for flow cytometric diagnosis and purification of HRS cells.
- To explore diagnostic markers on HRS cells and associated reactive T cells.
Main Methods:
- Utilized 6- and 9-color flow cytometric assays for CHL immunophenotyping.
- Employed established gating strategies for HRS cell population identification.
- Investigated HRS-T cell rosetting and immunophenotypic changes in reactive T cells.
Main Results:
- Achieved 85-90% sensitivity and nearly 100% specificity in diagnosing CHL by flow cytometry.
- Identified HRS-T cell rosetting as a diagnostically useful phenomenon.
- Observed characteristic immunophenotypic changes in reactive T cells associated with CHL.
Conclusions:
- Flow cytometry is a sensitive and specific method for diagnosing classical Hodgkin lymphoma.
- Flow cytometry enables the identification of HRS cells and associated reactive T cell changes.
- Flow cytometry facilitates rapid purification of HRS cells for further study.
Abstract:
Flow cytometry is an established technique to immunophenotype hematopoietic neoplasms. While the diagnosis of classical Hodgkin lymphoma (CHL) has commonly been made using paraffin sections, we have recently demonstrated that the neoplastic Hodgkin and Reed-Sternberg (HRS) cells of CHL can be identified by flow cytometry. Using 6- and 9-color flow cytometric assays, CHL can be immunophenotyped with 85-90% sensitivity and nearly 100% specificity. Analysis of this data requires using established gating strategies to help in the identification of putative HRS cell populations. Interestingly, HRS cells bind to reactive T cells (HRS-T cell rosetting) and this phenomenon can be identified and utilized diagnostically by flow cytometry. In addition, the reactive T cells of CHL show characteristic immunophenotypic changes by flow cytometry and these changes can suggest a diagnosis of CHL. Finally, these principles can be employed to rapidly purify HRS cells using flow cytometric cell sorting. This manuscript provides experimental protocols for immunophenotyping CHL by flow cytometry as well as purifying the HRS cells via flow cytometric cell sorting.
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