Clinicopathologic Features of CD5-positive Nodal Marginal Zone Lymphoma
Clinicopathologic Features of CD5-positive Nodal Marginal Zone Lymphoma
We searched the archives of our hospitals from January 1, 2000, through June 30, 2010, for cases of B-cell lymphoma that were positive for CD5 by immunohistochemistry, flow cytometric immunophenotyping, or both methods. We then selected cases of NMZL as defined using the criteria of the 2008 WHO classification. Clinical staging was based on the Ann Arbor system. The workup included history and physical examination; computerized tomographic scans of the neck, thorax, and abdomen; and upper gastrointestinal endoscopy in a subset of patients to exclude the presence of extranodal disease. Six of seven patients underwent bone marrow aspiration and biopsy; one patient declined examination. Clinical and laboratory data were obtained by review of medical records. The study was conducted according to an MD Anderson Cancer Center institutional review board–approved protocol.
We reviewed H&E-stained slides prepared from formalin-fixed, paraffin-embedded tissue sections from the diagnostic lymph node biopsy specimens of each patient. In six patients, Wright-Giemsa–stained bone marrow aspirate smears and touch imprints were reviewed, and 500-cell differential counts were performed. In addition, H&E-stained slides of bone marrow aspirate clot and biopsy specimens were reviewed.
Flow cytometric immunophenotypic analysis was performed on cell suspensions of lymph node, bone marrow aspirate, and peripheral blood specimens using a FACScan instrument (Becton-Dickinson Biosciences, San Jose, CA) as described previously. The lymphocyte population was gated using right angle side scatter and CD45 expression. The panel of monoclonal antibodies included reagents specific for CD3, CD5, CD10, CD19, CD20, CD22, CD23, CD38, CD43, FMC-7, and immunoglobulin κ and λ light chains (Becton-Dickinson).
Immunohistochemical analysis was performed using formalin-fixed, paraffin-embedded tissue sections; an avidin-biotin-peroxidase complex method; and an automated immunostainer (Ventana Biotech, Tucson, AZ), as described previously. All tissue sections underwent heat-induced antigen retrieval. The antibodies used included reagents specific for CD3 (Dako, Carpinteria, CA), CD5 (Labvision/Neomarkers, Fremont, CA), CD10 (Novocastra/Vision Biosystem, Newcastle upon Tyne, UK), CD20 (Dako), PAX-5 (Transduction Labs, San Diego, CA), BCL-2 (Novocastra/Vision Biosystem), BCL-6 (Dako), SOX11 (Cell Marque, Rocklin, CA), and cyclin D1 (SP4, Labvision/Neomarkers). CD5 expression was evaluated by comparing the pattern and intensity of staining with antibodies specific for CD3, CD5, and CD20. Neoplastic B cells typically expressed CD5 in a membranous pattern and with dimmer staining intensity compared with reactive T cells.
The results of conventional cytogenetics were available in four of six patients who underwent bone marrow aspiration and biopsy. Conventional cytogenetic analysis was performed on metaphase cells prepared from bone marrow aspirate specimens using standard techniques. Giemsa-banded metaphases were analyzed and the results were reported using the International System for Human Cytogenetic Nomenclature (2013).
We performed a second search of our hospital archives from January 1, 2000, through June 30, 2010, for CD5-negative cases of NMZL. Clinical and radiographic data were obtained from the medical record. Clinical staging and workup were similar to that performed for the patients with CD5-positive NMZL. Cases that showed evidence of extranodal or splenic disease at any point in the clinical course were excluded.
We identified 91 cases of CD5-negative NMZL. There were 34 men and 47 women, with a median age of 60 years (range, 21–87 years). Staging information was available for 66 patients. Frequency of bone marrow involvement and presence of lymphadenopathy above and below the diaphragm were compared with the CD5-positive NMZL group. This comparison was performed using a two-tailed Fisher exact test using GraphPad Prism software (version 6.01, GraphPad Software, La Jolla, CA). Findings with P ≤ .05 were considered statistically significant.
Materials and Methods
Case Selection and Morphologic Review
We searched the archives of our hospitals from January 1, 2000, through June 30, 2010, for cases of B-cell lymphoma that were positive for CD5 by immunohistochemistry, flow cytometric immunophenotyping, or both methods. We then selected cases of NMZL as defined using the criteria of the 2008 WHO classification. Clinical staging was based on the Ann Arbor system. The workup included history and physical examination; computerized tomographic scans of the neck, thorax, and abdomen; and upper gastrointestinal endoscopy in a subset of patients to exclude the presence of extranodal disease. Six of seven patients underwent bone marrow aspiration and biopsy; one patient declined examination. Clinical and laboratory data were obtained by review of medical records. The study was conducted according to an MD Anderson Cancer Center institutional review board–approved protocol.
We reviewed H&E-stained slides prepared from formalin-fixed, paraffin-embedded tissue sections from the diagnostic lymph node biopsy specimens of each patient. In six patients, Wright-Giemsa–stained bone marrow aspirate smears and touch imprints were reviewed, and 500-cell differential counts were performed. In addition, H&E-stained slides of bone marrow aspirate clot and biopsy specimens were reviewed.
Immunophenotypic and Cytogenetic Analysis
Flow cytometric immunophenotypic analysis was performed on cell suspensions of lymph node, bone marrow aspirate, and peripheral blood specimens using a FACScan instrument (Becton-Dickinson Biosciences, San Jose, CA) as described previously. The lymphocyte population was gated using right angle side scatter and CD45 expression. The panel of monoclonal antibodies included reagents specific for CD3, CD5, CD10, CD19, CD20, CD22, CD23, CD38, CD43, FMC-7, and immunoglobulin κ and λ light chains (Becton-Dickinson).
Immunohistochemical analysis was performed using formalin-fixed, paraffin-embedded tissue sections; an avidin-biotin-peroxidase complex method; and an automated immunostainer (Ventana Biotech, Tucson, AZ), as described previously. All tissue sections underwent heat-induced antigen retrieval. The antibodies used included reagents specific for CD3 (Dako, Carpinteria, CA), CD5 (Labvision/Neomarkers, Fremont, CA), CD10 (Novocastra/Vision Biosystem, Newcastle upon Tyne, UK), CD20 (Dako), PAX-5 (Transduction Labs, San Diego, CA), BCL-2 (Novocastra/Vision Biosystem), BCL-6 (Dako), SOX11 (Cell Marque, Rocklin, CA), and cyclin D1 (SP4, Labvision/Neomarkers). CD5 expression was evaluated by comparing the pattern and intensity of staining with antibodies specific for CD3, CD5, and CD20. Neoplastic B cells typically expressed CD5 in a membranous pattern and with dimmer staining intensity compared with reactive T cells.
The results of conventional cytogenetics were available in four of six patients who underwent bone marrow aspiration and biopsy. Conventional cytogenetic analysis was performed on metaphase cells prepared from bone marrow aspirate specimens using standard techniques. Giemsa-banded metaphases were analyzed and the results were reported using the International System for Human Cytogenetic Nomenclature (2013).
Comparison With CD5-negative NMZL Patients
We performed a second search of our hospital archives from January 1, 2000, through June 30, 2010, for CD5-negative cases of NMZL. Clinical and radiographic data were obtained from the medical record. Clinical staging and workup were similar to that performed for the patients with CD5-positive NMZL. Cases that showed evidence of extranodal or splenic disease at any point in the clinical course were excluded.
We identified 91 cases of CD5-negative NMZL. There were 34 men and 47 women, with a median age of 60 years (range, 21–87 years). Staging information was available for 66 patients. Frequency of bone marrow involvement and presence of lymphadenopathy above and below the diaphragm were compared with the CD5-positive NMZL group. This comparison was performed using a two-tailed Fisher exact test using GraphPad Prism software (version 6.01, GraphPad Software, La Jolla, CA). Findings with P ≤ .05 were considered statistically significant.
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