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Use of CD32, CD44 and CD71 to differentiate follicular lymphoma and diffuse large B-cell lymphoma subtypes by flow cytometry
  1. Lukas W Schwarz1,
  2. Jason E Love2,
  3. Kikkeri N Naresh3,
  4. Afshin Shameli2,
  5. Jonathan R Fromm2
  1. 1 University of Washington School of Medicine, Seattle, Washington, USA
  2. 2 Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA
  3. 3 Pathology / Cancer Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA
  1. Correspondence to Dr Jonathan R Fromm, Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA; jfromm{at}

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Many lymphomas can be immunophenotyped by flow cytometry (FCM), providing valuable diagnostic information.1 However, FCM cannot typically be used to distinguish follicular lymphoma (FL), diffuse large B-cell lymphoma, germinal centre type (DLBCL-GCB) and diffuse large B-cell lymphoma, non-germinal centre type (DLBCL-nGCB).

To expand the utility of clinical FCM, we compared the expression of CD32, CD44, CD71 and forward and side light scatter in a series of DLBCL-GCB, DLBCL-nGCB and FL specimens, and compared the results to all B cells in non-neoplastic lymph nodes (follicular hyperplasia, FH). These studies demonstrate that germinal centre (GC) B-cell lymphomas (FL and DLBCL-GCB) show decreased expression of CD44, while CD71 tends to be increased in DLBCL relative to FL. Both DLBCL-nGCB and DLBCL-GCB showed increased side light scatter relative to low-grade FL and FH.

The project was completed under the approval of the Human Subjects Division of the University of Washington. 5 DLBCL-nGCB, 12 DLBCL-GCB, 15 FL grade 1–2 (FL1-2), 10 FL grade 3 (FL3; including 7 grade 3A and 3 grade 3B cases, grouped, given the small number of cases) and 6 FH were evaluated. MFI (median fluorescence intensity) and MLI (median light intensity, analogue of MFI) for forward light scatter (FSC) and side light scatter (SSC) were analysed in GraphPad Prism (V.10). Evaluation of differences between diagnoses was performed using Tukey’s multiple comparison test.

Cryopreserved specimens were thawed and diluted to yield at least 150 000 cells (where possible) and stained with a standard stain-lyse-wash procedure (combination in online supplemental table 1) using a protocol described previously.1 FCM studies were performed on a 17-colour LSR II flow cytometer and the data analysed using Woodlist V.3.1.3 (courtesy of Dr Brent L Wood). MFI (each antigen) and MLI were collected …

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  • Handling editor Vikram Deshpande.

  • Contributors JRF conceived of the project, designed the experiments, analysed data and wrote the manuscript. LWS performed all experiments and analysed data. JEL, AS and KNN analysed data. All authors reviewed the manuscript.

  • Funding This work was funded by the Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.

  • Competing interests None declared.

  • Provenance and peer review Not commissioned; externally peer reviewed.

  • Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.