Immunophenotypic and gene expression analysis of monoclonal B-cell lymphocytosis shows biologic characteristics associated with good prognosis CLL

M C Lanasa, S D Allgood, S L Slager, S S Dave, C Love, G E Marti, N E Kay, C A Hanson, K G Rabe, S J Achenbach, L R Goldin, N J Camp, B K Goodman, C M Vachon, L G Spector, L Z Rassenti, J F Leis, J P Gockerman, S S Strom, T G Call, M Glenn, J R Cerhan, M C Levesque, J B Weinberg, N E Caporaso, M C Lanasa, S D Allgood, S L Slager, S S Dave, C Love, G E Marti, N E Kay, C A Hanson, K G Rabe, S J Achenbach, L R Goldin, N J Camp, B K Goodman, C M Vachon, L G Spector, L Z Rassenti, J F Leis, J P Gockerman, S S Strom, T G Call, M Glenn, J R Cerhan, M C Levesque, J B Weinberg, N E Caporaso

Abstract

Monoclonal B-cell lymphocytosis (MBL) is a hematologic condition wherein small B-cell clones can be detected in the blood of asymptomatic individuals. Most MBL have an immunophenotype similar to chronic lymphocytic leukemia (CLL), and 'CLL-like' MBL is a precursor to CLL. We used flow cytometry to identify MBL from unaffected members of CLL kindreds. We identified 101 MBL cases from 622 study subjects; of these, 82 individuals with MBL were further characterized. In all, 91 unique MBL clones were detected: 73 CLL-like MBL (CD5(+)CD20(dim)sIg(dim)), 11 atypical MBL (CD5(+)CD20(+)sIg(+)) and 7 CD5(neg) MBL (CD5(neg)CD20(+)sIg(neg)). Extended immunophenotypic characterization of these MBL subtypes was performed, and significant differences in cell surface expression of CD23, CD49d, CD79b and FMC-7 were observed among the groups. Markers of risk in CLL such as CD38, ZAP70 and CD49d were infrequently expressed in CLL-like MBL, but were expressed in the majority of atypical MBL. Interphase cytogenetics was performed in 35 MBL cases, and del 13q14 was most common (22/30 CLL-like MBL cases). Gene expression analysis using oligonucleotide arrays was performed on seven CLL-like MBL, and showed activation of B-cell receptor associated pathways. Our findings underscore the diversity of MBL subtypes and further clarify the relationship between MBL and other lymphoproliferative disorders.

Figures

Figure 1. Immunophentype profiling in MBL
Figure 1. Immunophentype profiling in MBL
In frames A through I, the MFI for each MBL case is shown for each cell surface marker. CLL-like MBL are designated with blue diamonds, atypical MBL are green triangles, and CD5NEG MBL are shown as blue squares. A. CD5 by CD20; B. CD5 by CD23; C. CD5 by sIg, D. CD5 by CD49d, E. CD5 by CD79b, F. CD5 by FMC7, G. IgD by IgM. To characterize the B cell maturation state of different MBL subtypes, plots of IgD by CD38 (H) and IgD by CD27 (I) were displayed.
Figure 2. Gene Expression Array Analysis and…
Figure 2. Gene Expression Array Analysis and ROR1 Expression in MBL
A. Gene set enrichment analysis of gene pathways comparing normal memory B cells (NL), CLL-like MBL cells, and CLL cells. Relative expression of differentially expressed genes is shown where green denotes lower expression and red shows increased expression. MAPKinase (MAPK), protein kinase A (PKA), and proliferation signatures show differential expression between groups. B. Quantile box plots of MFI ROR1 in MBL and residual normal CD19+ B lymphocytes. CLL-like MBL showed greater surface expression of ROR1 than either CD5neg MBL or normal B cells (p< 0.005, t-test; designated **). Atypical MBL showed greater surface expression than normal B cells.

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