B-cell chronic lymphocytic leukemia cells express a surface membrane phenotype of activated, antigen-experienced B lymphocytes by Rajendra N. Damle, Fabio.

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B-cell chronic lymphocytic leukemia cells express a surface membrane phenotype of activated, antigen-experienced B lymphocytes by Rajendra N. Damle, Fabio Ghiotto, Angelo Valetto, Emilia Albesiano, Franco Fais, Xiao-Jie Yan, Cristina P. Sison, Steven L. Allen, Jonathan Kolitz, Philip Schulman, Vincent P. Vinciguerra, Petra Budde, Jurgen Frey, Kanti R. Rai, Manlio Ferrarini, and Nicholas Chiorazzi Blood Volume 99(11):4087-4093 June 1, 2002 ©2002 by American Society of Hematology

Expression of surface membrane markers on B-CLL cells and normal CD5+ B cells.Surface membrane markers overexpressed on B-CLL cells as compared with normal CD5+ B cells. Expression of surface membrane markers on B-CLL cells and normal CD5+ B cells.Surface membrane markers overexpressed on B-CLL cells as compared with normal CD5+ B cells. PBMCs from B-CLL cases and age-matched healthy donors were analyzed for marker expression on CD5+CD19+ cells by means of 3-color immunofluorescence. (A) Representative flow cytometric profiles of expression of these markers on B cells from a 67-year-old healthy donor (N39) and a B-CLL patient (CLL 308). (B) Paired bars depict the mean ± SE of the percentage of CD5+CD19+cells expressing these markers in 61 (51 IgM+ and 10 IgG+) B-CLL patients and 20 age-matched healthy controls studied. B-CLL cells express significantly higher percentages (P < .0001) of CD27, CD39, CD23, CD25, CD69, and CD71 than normal CD5+CD19+ cells. The expression of these markers was variable both in the patients and in the markers, with the greatest variability seen for CD25. The SE and the SD for these markers ranged from 1.7 to 3.6 and from 13.1 to 28.0, respectively. Rajendra N. Damle et al. Blood 2002;99:4087-4093 ©2002 by American Society of Hematology

Expression of surface membrane markers on B-CLL cells and normal CD5+ B cells.Surface membrane markers underexpressed on B-CLL cells as compared with normal CD5+ B cells. Expression of surface membrane markers on B-CLL cells and normal CD5+ B cells.Surface membrane markers underexpressed on B-CLL cells as compared with normal CD5+ B cells. PBMCs from B-CLL patients and age-matched healthy donors were analyzed for marker expression on CD5+CD19+ cells. (A) Representative flow cytometric profiles of expression of these markers on B cells from a 67-year-old healthy donor (N39) and CLL 308. (B) Paired bars depict the mean ± SE of the percentage of CD5+CD19+ cells expressing these markers in 51 IgM+ and 10 IgG+ B-CLL patients and 20 age-matched healthy controls. B-CLL cells expressed significantly lower percentages (P < .0001) of CD40, CD72, CD22, CD79b, CD62L, CD38, FcγRIIb, CD40L, CD95, and CD80 than normal CD5+CD19+ cells. The expression of these markers was variable both in the patients and in the markers, with the greatest variability seen for CD38. The SE and the SD for these markers ranged from 1.2 to 4.7 and from 8.9 to 35.5, respectively. Rajendra N. Damle et al. Blood 2002;99:4087-4093 ©2002 by American Society of Hematology

Comparison of density of expression of surface membrane markers by B-CLL cells and normal CD5+ B cells.The density of expression of CD5, CD27, CD38, CD22, FcγRIIb, IgD, and IgM on B-CLL cells was compared with that on CD5+CD19+ cells from age-matched contro... Comparison of density of expression of surface membrane markers by B-CLL cells and normal CD5+ B cells.The density of expression of CD5, CD27, CD38, CD22, FcγRIIb, IgD, and IgM on B-CLL cells was compared with that on CD5+CD19+ cells from age-matched controls. The density of expression of a membrane molecule was estimated as a ratio of specific-to-nonspecific binding of isotype “control” antibody and is expressed as the MFIR on the y-axis. Dark lines represent staining of cells using antibodies to the molecules listed whereas the thin lines represent the staining with isotype-matched control antibodies. (A) Representative flow cytometric profiles of expression of these markers on CD5+CD19+ B cells from a 67-year-old healthy donor (N39) and CLL 308. (B) Paired bars depict the mean ± SE of the MFIR of expression of these markers on CD5+CD19+ cells in 51 IgM+ B-CLL patients and 20 age-matched healthy controls. Although the IgG+ B-CLL patients expressed CD5, CD27, CD38, CD22, and FcγRIIb at densities identical to those of IgM+B-CLL patients, the data in this Figure do not include the IgG+ B-CLL patients since they do not express either smIgM (smIgM) or smIgD. All differences were statistically significant (P < .001). Rajendra N. Damle et al. Blood 2002;99:4087-4093 ©2002 by American Society of Hematology

Comparison of surface membrane phenotypes of IgM+ B-CLL cases grouped on the basis of immunoglobulin V gene mutation.PBMCs from 51 IgM+ B-CLL cases were analyzed for the expression of CD38, CD39, CD40, CD62L, CD69, and CD71 on CD5+CD19+ cells. Comparison of surface membrane phenotypes of IgM+ B-CLL cases grouped on the basis of immunoglobulin V gene mutation.PBMCs from 51 IgM+ B-CLL cases were analyzed for the expression of CD38, CD39, CD40, CD62L, CD69, and CD71 on CD5+CD19+ cells. The B-CLL cases were segregated into 2 groups: unmutated, in which the expressed immunoglobulin V genes showed less than a 2.0% difference from the most similar germ line gene, and mutated, in which the expressed immunoglobulin V genes showed a 2.0% or greater difference from the most similar germ line. The mean ± SE of the percentages of CD5+CD19+ cells expressing these markers in the unmutated and mutated cases are represented. All differences were statistically significant (P < .001). Rajendra N. Damle et al. Blood 2002;99:4087-4093 ©2002 by American Society of Hematology

Comparison of density of HLA-DR expression on B-CLL cells and normal CD5+ B cells.PBMCs from 61 B-CLL cases and 20 age-matched normal donors were analyzed for the expression of HLA-DR on CD5+CD19+ cells. Comparison of density of HLA-DR expression on B-CLL cells and normal CD5+ B cells.PBMCs from 61 B-CLL cases and 20 age-matched normal donors were analyzed for the expression of HLA-DR on CD5+CD19+ cells. Bars represent the mean ± SE of the MFIR of HLA-DR expression on CD5+CD19+ cells. The differences between the unmutated and mutated subgroups and between the CD38+and the CD38− subgroups are significant (P < .001). In addition, the differences between the unmutated subgroup and the normal CD5+ B cells, and between the CD38+ subgroup and the normal CD5+ B cells, are also significant (P < .001). Rajendra N. Damle et al. Blood 2002;99:4087-4093 ©2002 by American Society of Hematology

Expression of CD69 and CD71 by B-CLL cells from unmutated and mutated cases.PBMCs from 61 B-CLL cases were analyzed specifically for the expression of CD69 and CD71 on CD5+CD19+ cells. Expression of CD69 and CD71 by B-CLL cells from unmutated and mutated cases.PBMCs from 61 B-CLL cases were analyzed specifically for the expression of CD69 and CD71 on CD5+CD19+ cells. The percentages of CD5+CD19+ cells expressing CD69 or CD71 were plotted independently in each of the 2 adjacent scattergrams. The symbol representing the percentage of cells expressing CD69 is connected with a line to the symbol representing the percentage of cells expressing CD71 in each individual unmutated (left panel) and mutated case (right panel). Overall, the percentages of cells expressing CD69 inversely correlated with those expressing CD71 (P < .001). Rajendra N. Damle et al. Blood 2002;99:4087-4093 ©2002 by American Society of Hematology

Comparison of surface membrane phenotypes of IgM+ B-CLL cases grouped on the basis of CD38 expression.PBMCs from 51 IgM+ B-CLL cases were analyzed for the expression of CD38, CD40, CD69, CD79b, and HLA-DR on CD5+CD19+ cells. Comparison of surface membrane phenotypes of IgM+ B-CLL cases grouped on the basis of CD38 expression.PBMCs from 51 IgM+ B-CLL cases were analyzed for the expression of CD38, CD40, CD69, CD79b, and HLA-DR on CD5+CD19+ cells. The B-CLL cases were segregated into 2 groups: CD38+, in which 30% or more CD5+CD19+ B cells expressed CD38, and CD38−, in which 30% or fewer CD5+CD19+ B cells expressed CD38. The mean ± SE of the percentages of CD5+CD19+ cells expressing CD40, CD69, or CD79b in the CD38+ and CD38− cases are provided. Rajendra N. Damle et al. Blood 2002;99:4087-4093 ©2002 by American Society of Hematology