Ann Lab Med.  2015 Mar;35(2):187-193. 10.3343/alm.2015.35.2.187.

Bone Marrow Flow Cytometry in Staging of Patients With B-cell Non-Hodgkin Lymphoma

• Kim B
• Lee ST
• Kim HJ
• Kim SH

• KMID: 2363177
• DOI: http://doi.org/10.3343/alm.2015.35.2.187

Abstract

BACKGROUND
Bone marrow biopsies are routinely performed for staging patients with B-cell non-Hodgkin lymphoma (NHL). In addition to histomorphological studies, ancillary tools may be needed for accurate diagnosis. We investigated the clinical utility of multiparameter flow cytometric examination of bone marrow aspirates.
METHODS
A total of 248 bone marrow specimens from 232 patients diagnosed with B-cell NHL were examined. Monoclonal antibodies directed against CD19, CD20, CD10 (or CD5), and kappa and lambda immunoglobulins were used. Multi-stage sequential gating was performed to select specific cells of interest, and the results were compared with bone marrow histology.
RESULTS
The concordance rate between histomorphology and flow cytometry was 91.5% (n=227). Eight cases (3.2%) were detected by flow cytometry alone and were missed by histomorphology analysis, and 6 of these 8 cases showed minimal bone marrow involvement (0.09-2.2%). The diagnosis in these cases included large cell lymphoma (n=3), mantle cell lymphoma (n=3), and mucosa-associated lymphoid tissue (MALT) lymphoma (n=2). Thirteen cases were histopathologically positive and immunophenotypically negative, and the diagnoses in these cases included diffuse large cell lymphoma (n=7), T-cell/histiocyte-rich large B-cell lymphoma (n=2), anaplastic lymphoma kinase (ALK)-positive large B-cell lymphoma (n=1), follicular lymphoma (n=1), MALT lymphoma (n=1), and unclassifiable lymphoma (n=1).
CONCLUSIONS
Multi-color flow cytometry can be a useful method for assessing bone marrow in staging NHL and also plays a complementary role, especially in detecting small numbers of lymphoma cells.

Keyword

Bone marrow; Immunophenotyping; Flow cytometry; Non-Hodgkin lymphoma

MeSH Terms

Antibodies, Monoclonal/immunology
Antigens, CD19/immunology/metabolism
Antigens, CD20/immunology/metabolism
Bone Marrow/*pathology
Female
Flow Cytometry
Humans
Immunophenotyping
Lymphoma, B-Cell/*pathology
Male
Neoplasm Staging
Neprilysin/immunology/metabolism
Antibodies, Monoclonal
Antigens, CD19
Antigens, CD20
Neprilysin

Figure

• 

  Fig. 1 Examples of κ-to-λ ratios for patients with B-cell non-Hodgkin lymphoma using different gating strategies. (A) Plots showing unequivocal light chain restriction. (B) Increased sensitivity using multi-color antibodies. Gating on CD19-positive and CD20-positive and CD10-negative cells could identify very small amounts of clonal B-cells (κ-to-λ ratio=11.5) (C) whereas gating without CD10 masks the clonal B-cell population (κ-to-λ ratio=1.412). (D) Plots showing negative controls.

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