D Cyclins in CD5+ B-Cell Lymphoproliferative Disorders

Abstract

We analyzed protein expression of cyclin D1, cyclin D2, and cyclin D3 using high-resolution enzymatic amplification staining and flow cytometry in the neoplastic cells from 80 patients with CD5+ B-cell lymphoproliferative disorders. The D cyclins were expressed differentially in chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), and mantle cell lymphoma (MCL) with strong staining of cyclin D1 and D2 in MCL, strong staining of cyclin D1 but weak staining of cyclin D2 in 4 of 5 PLLs, and low-level staining for both cyclins in most CLLs. No correlation between cyclin D1 and D2 and growth rates or CD38 expression was observed. However, cyclin D1 levels were significantly higher in ZAP-70+ CLL cases, although no association between ZAP-70 and cyclin D2 was detected. The results indicate that flow cytometric analysis of D cyclins may help in classification of CD5+ B-cell lymphoproliferative disorders.

Flow cytometric immunophenotyping for hematologic neoplasms

“…Although several groups have attempted to develop a flow cytometric assay for cyclin-D1, most lack sensitivity, and probably the most sensitive method, which uses an enzymatic amplification step, appears to lack specificity in the distinction between CLL/SLL and MCL. 31 MZL is CD5 ϩ in approximately 5% of patients and can be difficult to distinguish from CLL/SLL with a variant phenotype and other CD5 ϩ lymphoid neoplasms. 32,33 Possible distinguishing features include lack of expression of CD23 in most MZL and presence of plasmacytic differentiation in a significant subset of MZL, as demonstrated by expression of CD138, bright expression of CD38, and cytoplasmic immunoglobulin light chain restriction in at least a subset of the neoplastic cells.…”

Correlation between ZAP‐70, phospho‐ZAP‐70, and phospho‐Syk expression in leukemic cells from patients with CLL

“…The samples were analyzed for the expression of ZAP-70, phospho-ZAP-70, phospho-Syk, and p21cip1 by Pathfinder Biotech using enzymatic amplification staining (EAS) as previously described (10,(12)(13)(14). EAS is a validated, catalyzed reporter deposition technology based on the enzymatic activity of peroxidase.…”

“…To assess how basal levels of phosphorylation of these molecules relate to levels of ZAP-70 without in vitro stimulation, we used a high resolution immunophenotyping technology with flow cytometric analysis (10) to assess ZAP-70, phospho-ZAP-70, and phosphoSyk in CLL cells directly ex vivo.…”

Cell‐type specific molecular expression levels by restricted‐dimensional cytometry

“…Co‐stains were obtained from BioLegend (San Diego, CA). Amplification staining was accomplished as previously described 15–28 . Briefly, the cells were incubated with primary antibodies at room temperature and washed thoroughly.…”

“…We used an amplification technology to enhance the specific analyte signals. [15][16][17][18][19][20][21][22][23][24][25][26][27][28] The paradigm for rdC is the assessment of only one lineage marker and one amplified analyte in each tube. In that way data analysis was significantly simplified.…”