CD38 expression in B-chronic lymphocytic leukemia: association with clinical presentation and outcome in 155 patients.

Department of Clinical Hematology, Institut Català d'Oncologia and Hospital Princeps d'Espanya, Ciutat Sanitaria i Universitaria de Bellvitge, Barcelona, Spain. 31577edd@comb.es

BACKGROUND AND OBJECTIVES: To investigate whether CD38 expression at diagnosis is an independent predictor of survival and assess its associations with other clinical parameters used in the staging of B-cell chronic lymphocytic leukemia (B-CLL). DESIGN AND METHODS: CD38 expression was analyzed in 155 consecutive unselected patients newly diagnosed with B-CLL from January 1991 to July 1997. In all cases CD38 expression was evaluated at diagnosis and patients were classified into two groups: those with > or = 30% were considered positive (CD38+) and those with < 30% were considered negative (CD38-). Statistical differences between each group were analyzed using c2 tests for categorical variables and Student's t-tests for continuous variables. Survival analysis was performed at the univariate level by the Kaplan Meier technique and at the multivariate level by Cox hazard models. RESULTS: Thirty (19%) patients were CD38+. CD38+ was associated with atypical morphology (p=0.004), a diffuse bone marrow pattern (p=0.016), Rai stage > or =2 (p=0.009), high lactate dehydrogenase (p=0.02), high b2 microglobulin (p=0.004), and higher lymphocyte count (p=0.02). Furthermore, CD38+ patients required treatment more frequently (p=0.006) and CLL-related mortality was significantly higher (p=0.012). When we divided the study group into patients with Rai 0-1 and Rai 2-4 stages, CD38 positivity was only significantly associated with mortality in the early stage patients (p= 0.012 vs p= 0.68). CD38 expression in the multivariate analysis lost its statistical significance. None of these results was modified when the CD38 cut-off was set at 20%. INTERPRETATION AND CONCLUSIONS: CD38 expression identifies a subgroup of B-CLL patients with aggressive clinical presentation and worse outcome. Its expression is probably associated with other prognostic factors, but the feasibility of determining this parameter makes it easily reproducible and adds prognostic information at diagnosis to aid prediction of the clinical course and outcome of B-CLL.

PMID: 12368155 [PubMed - indexed for MEDLINE]

Significance of zeta-associated protein (ZAP-70) and CD38 expression in chronic lymphocytic leukemia.

Clinical Pathology Department, Faculty of Medicine, Tanta University, Egypt.

Chronic lymphocytic leukemia (CLL) is a disease with a highly variable clinical course; some patients never need treatment, while others require intensive treatment early after diagnosis. Some new prognostic factors, such as immunoglobulin variable heavy chain (IgVH) mutational status, zeta-associated protein (ZAP-70) and CD38 expression in leukemic cells were introduced to identify attenuated versus progressive types of CLL bearing the potential to facilitate risk-adapted treatment strategies. So, the aim of this work is to evaluate the clinical value of ZAP-70 and CD38 as predictors of disease progression. We assessed the expression of these markers by flowcytometry in 38 patients with CLL and correlated their levels with genetic abnormalities detected by fluorescence in situ hybridization (FISH) and the clinical outcome. We found that 18 patients (47.4 %) were positive for ZAP-70 (> or = 20%) and 16 patients (42.1%) were positive for CD38 (> or = 20%). Positive ZAP-70 and CD38 status were associated with an unfavorable clinical course including high leukocytic count, lymphocytosis, high lactate dehydrogenase (LDH) serum level, advanced disease stage, trisomy 12 and del (11q); negative ZAP-70 and CD38 status were correlated with del (13q). The treatment-free survival time was 30 months for ZAP-70-positive patients and 18 months for ZAP-70-nagative patients (p < 0.01). Combined analysis of ZAP-70 and CD38 yielded discordant results in 10 patients (26.3 %), whereas 16 patients (42.1%) were concordantly negative and 12 patients (31.6%) were concordantly positive for ZAP-70 and CD38 expression. Median treatment-free survival times in patients whose leukemic cells were ZAP-70+CD38+ was 27 months as compared to 100 months in patients with a ZAP-70(-)CD38(-) status. In patients with discordant ZAP-70/CD38 results, the median treatment-free survival time was 40 months. Thus, ZAP-70 and CD38 expression analyses provide complementary prognostic information and allow distinguishing the patients groups with the most favorable prognosis as well as those with the worst. The current findings suggest that both ZAP-70 and CD38 protein expression should be assessed in patients with CLL for the definition of prognostic subgroups.

PMID: 18689273 [PubMed - indexed for MEDLINE]

Combined analysis of ZAP-70 and CD38 expression as a predictor of disease progression in B-cell chronic lymphocytic leukemia.

Department of Hematology and Oncology, Georg-August-University Goettingen, Germany. jan.duerig@uni-essen.de

Prognostic predictions in B-cell chronic lymphocytic leukemia (B-CLL) at early clinical stage are based on biological disease parameters, such as ZAP-70 and CD38 protein levels, genomic aberrations as well as immunoglobulin variable heavy chain gene (IgV(H)) mutation status. In the current study, ZAP-70 and CD38 expressions were examined by flow cytometry in 252 patients with B-CLL. Cytoplasmic ZAP-70 expression in more than 20% (ZAP-70(+)) and surface CD38 expression on more than 30% (CD38(+)) of B-CLL cells were associated with an unfavorable clinical course. The levels of ZAP-70 and CD38 did not change over time in the majority of patients where sequential samples were available for analysis. Combined analysis of ZAP-70 and CD38 yielded discordant results in 73 patients (29.0%), whereas 120 patients (47.6%) were concordantly negative and 59 patients (23.4%) were concordantly positive for ZAP-70 and CD38 expression. Median treatment-free survival times in patients whose leukemic cells were ZAP-70(+)CD38(+) was 30 months as compared to 130 months in patients with a ZAP-70(-)CD38(-) status. In patients with discordant ZAP-70/CD38 results, the median treatment-free survival time was 43 months. Thus, ZAP-70 and CD38 expression analyses provided complementary prognostic information identifying three patient subgroups with good, intermediate and poor prognosis. Over-representation of high-risk genomic aberrations such as 17p deletion or 11q deletion and distribution of the IgV(H) mutation status in B-CLL discordant for ZAP-70/CD38 pointed toward a distinct biologic background of the observed disease subgroups. This finding was also supported by microarray-based gene expression profiling in a subset of 35 patients. The expression of 37 genes differed significantly between the three groups defined by their expression of ZAP-70 and CD38, including genes that are involved in regulation of cell survival and chemotherapy resistance.

PMID: 15759031 [PubMed - indexed for MEDLINE]

Immunophenotypic characterization of the leukemic B-cells from Iranian patients with chronic lymphocytic leukemia: association between CD38 expression and disease progression.

Department of Immunology, School of Public Health, Medical Sciences/Tehran University, Tehran, Iran.

BACKGROUND: Patients with B-cell chronic lymphocytic leukemia (B-CLL) have heterogeneous clinical courses, thus several biological parameters need to be added to the current clinical staging systems to predict disease outcome. Recent immunophenotypic studies performed mainly in Western populations have demonstrated the prognostic value of CD38 and ZAP-70 expression in B-CLL. OBJECTIVES: To investigate the expression pattern of a variety of membrane antigens on leukemic cells from Iranian patients with CLL and to find out if there are any differences in the expression of these markers between indolent and progressive groups. METHODS: In the present study, peripheral blood samples from 87 Iranian patients with B-CLL were analysed by flow cytometry. RESULTS: In all cases, the neoplastic cells displayed B-CLL phenotype (CD5+/CD19+/sIg+). The vast majority of the cases expressed CD23, but failed to stain for CD3 or CD14. The leukemic cells of most patients expressed CD27 (84/87, 95.4%) and CD45RO (74/87, 83.9%) molecules, suggesting a memory B-cell phenotype. Comparison between the indolent (n=42) and progressive (n=37) patients revealed significantly higher frequency and intensity of CD38 expression in progressive group (40.5%) compared to indolent (11.9%) patients (p<0.05). None of the other membrane antigens were differentially expressed in these two groups of patients. CONCLUSION: Our results obtained in an Asian ethnic population confirm and extend previous findings obtained from Western populations regarding the association of CD38 expression and disease progression in B-CLL.

PMID: 18319522 [PubMed - indexed for MEDLINE]

ZAP-70 protein expression and CD38 positivity in B-cell chronic lymphocytic leukemia.

Institut Universitaire d'Hématologie, Hopital St Louis, 75010 Paris, France.

BACKGROUND: The clinical course, disease progression, and survival of B-cell chronic lymphocytic leukemia (B-CLL) have been correlated with immunoglobulin heavy-chain variable region mutation status. The biologic parameters 70-kDa zeta-associated protein (ZAP-70) and CD38 expression are easier and faster surrogate markers for mutational status. OBJECTIVE: To assess retrospectively ZAP-70 expression in B-CLL cells using flow cytometry and examine its relationship with CD38 expression and the median time from diagnosis to initial therapy. METHODS: Ninety-four unselected patients who had their follow-up in the outpatient clinic from 2004 to 2005 were reviewed for immunophenotyping ZAP-70 and CD38 expression. Direct immunolabeling with clone 2E3.2, isotype IgG2a, enabled easy quantification of ZAP-70 by flow cytometry in association with CD38 expression; in addition, the mean fluorescence intensity ratio (MFIR) of CD19+CD5+ B-CLL cells compared to an isotype control monoclonal antibody was determined. RESULTS: ZAP-70 expression levels in B-CLL cells varied widely (0.3-99%). The median time to therapy was significantly shorter for the 54 patients with 20% or more ZAP-70+ cells (30 months) than for the 40 patients with less than 20% ZAP-70+ cells (median time to treatment not reached). The optimal MFIR for classifying patients as ZAP-70+ was 2. Thirty-two patients had a threshold of ZAP-70+CD38+ greater than 30%, with a median time from diagnosis to treatment of 19 months. Regardless of CD38 expression level, CD38 and ZAP-70 expressions were significantly associated. The median interval from diagnosis to initial therapy was 16.2 months for ZAP-70+CD38+ patients, 60 months for ZAP-70+CD38- or ZAP-70-CD38+ patients, and had not yet been reached for ZAP-70-CD38- patients. CONCLUSION: The association of ZAP-70+CD19+CD5+ B-CLL cells and percentage of CD38+CD19+CD5+ B-CLL cells evaluated by flow cytometry provide reliable methods that could be introduced into a routine diagnostic B-CLL panel to predict outcome.

PMID: 18322442 [PubMed - indexed for MEDLINE]

Immunophenotypic features distinguishing familial chronic lymphocytic leukemia from sporadic chronic lymphocytic leukemia.

Good Samaritan Hospital, Dayton, Ohio, USA.

BACKGROUND: Familial chronic lymphocytic leukemia (CLL) has the most frequent familial aggregation among hematological malignancies. Familial CLL families have been studied to identify susceptibility genes and other factors that contribute in the etiology of CLL. To date no study has been conducted to evaluate and compare patterns of cell surface antigen expression in familial CLL and sporadic CLL. METHODS: The pattern of cell surface antigen expression was studied in familial and sporadic CLL to determine if unique identifiers of familial CLL could be detected. Survival in familial CLL verses sporadic CLL was compared and the association between prognosis and CD38 expression studied. RESULTS: Familial and sporadic CLL demonstrated the same characteristic immunophenotype (positive for surface immunoglobulin, CD5, CD19, and CD23 with dim CD20, and CD22). CD2 and CD13 expression, however, were more frequent (30% of cases) in familial CLL (P = 0.0003 for CD2, P = 0.006 for CD13) than in sporadic CLL (2-6%). There was no significant difference in survival in the two groups studied. Although the incidence of CD38 expression was similar in familial and sporadic CLL (47% and 44% respectively) the association with prognosis differed. There was a trend to decreased survival in CD38 positive sporadic (P = 0.06) but not familial CLL patients. CONCLUSIONS: We conclude that detection of CD2 or CD13 expression in CLL suggests familial CLL and examination of family history for additional affected members is warranted. Furthermore, CD38 expression does not carry the negative prognosis observed in sporadic CLL.

PMID: 18431797 [PubMed - indexed for MEDLINE]

IgVH genes mutation and usage, ZAP-70 and CD38 expression provide new insights on B-cell prolymphocytic leukemia (B-PLL).

Section of Haemato-Oncology--Institute of Cancer Research, Sutton, UK. idelgiu@freemail.it

B-prolymphocytic leukemia (B-PLL) is a rare disease with poor prognosis. To further characterize the biological features of this disease, we analyzed immunoglobulin heavy chain (IgVH) mutations, ZAP-70 and CD38 in 19 cases with de novo B-PLL. Immunoglobulin heavy chain genes analysis showed an unmutated pattern (>98% homology to germ line) in 9/17 cases (53%), with 100% homology in eight. In the remaining, it ranged from 90 to 97.4%, with three cases slightly mutated (98-95%) and five heavily mutated (<95%). All B-PLL utilized members of VH3 (11/17) and VH4 (6/17) families, with V3-23, V4-59 and V4-34 gene accounting for more than half of them, regardless of mutational status. ZAP-70, assessed by flow cytometry, ranged from 1 to 91% cells, being > or =20% in 57% of cases. CD38 ranged from 1 to 99% (median 21%). There was no correlation between IgVH status and ZAP-70 or CD38 expression, but male gender and del(17p) were more common in the unmutated group. Neither IgVH mutations, CD38 expression nor del(17p) influenced patients' outcome. Unexpectedly, ZAP-70+ B-PLL patients survived longer (40 months) than ZAP-70- B-PLL (8 months). B-PLL appears biologically heterogeneous regarding IgVH mutations, ZAP-70 and CD38 expression, showing a pattern distinct from that of other lymphoproliferative disorders.

PMID: 16642047 [PubMed - indexed for MEDLINE]

Zap-70 and CD38 as predictors of IgVH mutation in CLL.

Department of Pathology, University of Mississippi Medical Center, 2500 North State Street, Jackson, MS 39216, USA.

Zeta-chain (TCR)-associated protein kinase 70 kDa (Zap-70) and CD38 expression may be of prognostic significance in chronic lymphocytic leukemia (CLL). Previous studies indicate that Zap-70 and CD38 are usually positive in cases of CLL with unmutated immunoglobulin variable region genes (IgVH) and may be used to predict IgVH mutation status and prognosis. Usually cases of CLL positive for Zap-70 or CD38 indicate a worse prognosis. In the present investigation, 47 cases of CLL were evaluated for CD38 expression, and 17 cases were evaluated for both Zap-70 and CD38 expression. Of the 47 cases, 19 (40.4%) positively expressed CD38. Of the 17 cases evaluated for Zap-70, 11 (64.7%) were positive for Zap-70, while only 6 (35.3%) were positive for CD38 expression; the remaining cases were negative for CD38. The results of this study show that Zap-70 expression may be a better indicator of the mutational status of IgVH and prognosis of CLL than CD38 expression. In addition, CD38 negativity does not necessarily indicate that IgVH mutation has occurred. These data point to the need for a more extensive study to evaluate the significance of Zap-70 and CD38 expression as indicators of IgVH mutation status and prognosis of CLL patients.

PMID: 17931624 [PubMed - indexed for MEDLINE]

ZAP-70 and CD38 expression are independent prognostic factors in patients with B-cell chronic lymphocytic leukaemia and combined analysis improves their predictive value.

Department of Haematology and Bone Marrow Transplantation, Medical University of Lublin. iwohus@wp.pl

Recently identified biological risk factors in B-cell chronic lymphocytic leukemia (B-CLL) include ZAP-70 and CD38 expression. The present study was conducted to clarify whether a combined analysis could improve predictive impact of these two parameters. We examined the expression of ZAP-70 and CD38 by flow cytometry method in 217 newly diagnosed, consecutive, unselected and well characterized B-CLL patients in relation to laboratory parameters and clinical outcome. We confirmed that both ZAP-70 as well as CD38 were independent of prognostic factors. There was a significant correlation between the percentage of leukemic cells positive for ZAP-70 and the percentage of CD38+CD19+ cells (R=0.629; p=0.000001). Combined analysis of ZAP-70 and CD38 showed concordant results in 158/217 patients (72.8%), while in 59 patients the results were discordant (27.2%). A mean treatment free survival (TFS) was the longest in ZAP-70-CD38-patients (45.6 months, comparing to 13.6 months in ZAP-70+CD38+ group). Also a mean overall survival was the longest in ZAP-70-CD38- patients (224.7 months compared to 77.9 months in ZAP-70+CD38+ patients).

PMID: 18519230 [PubMed - indexed for MEDLINE]

Regulation of CD38 in proliferating chronic lymphocytic leukemia cells stimulated with CD154 and interleukin-4.

Division of Investigative Sciences, Department of Haematology, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK.

BACKGROUND AND OBJECTIVES: Chronic lymphocytic leukemia (CLL) cells, like normal B-cells, exist in two populations in vivo: quiescent cells in the peripheral circulation and proliferating cells in lymph nodes. The surface marker CD38 has roles in cell adhesion and signaling. Its expression correlates with poor clinical outcome and is associated with expression of the signaling intermediate ZAP-70, which is also a marker of poor prognosis. We investigated the regulation of CD38 and ZAP-70 in proliferating CLL cells. DESIGN AND METHODS: We cultured CLL cells on a stromal cell layer that maintains viability and also with some stromal cells expressing CD40 ligand (CD154) in order to measure changes in expression of CD38 and ZAP-70. RESULTS: We demonstrated up-regulation of CD38 expression by CD154. The degree of up-regulation did not correlate with clinical stage or mutational status. In addition in the majority of cases tested ZAP-70 expression increased in parallel with up-regulation of CD38 although discordant cases were also observed. INTERPRETATION AND CONCLUSIONS: Overall we demonstrated that regulation of CD38 in CLL is dynamic and dependent on signals from CD154 and a stromal cell layer. We speculate that CD38 and ZAP-70 are expressed in lymph node leukemic cells in both good and poor prognosis patients, but, in cases with good clinical outcome, these molecules are down-regulated in the peripheral blood whereas in cases with poor prognosis their expression is maintained.

PMID: 18024373 [PubMed - indexed for MEDLINE]

Prognostic significance of combined analysis of ZAP-70 and CD38 in chronic lymphocytic leukemia.

Hematology Oncology Unit, National Cancer Institute, Fondazione G. Pascale, Via Mariano Semmola, Naples, Italy. giovannidarena@libero.it

The clinical heterogeneity that characterizes chronic lymphocytic leukemia (CLL) poses critical questions concerning the identification of high risk patients. Unmutated IgV(H) genes, CD38 and ZAP-70 expression have emerged as the most useful tools in identifying aggressive CLL. The simultaneous expression of ZAP-70 and CD38 in 157 patients with CLL has been evaluated. Fifty-seven patients (36%) were positive for ZAP-70 and 46 patients (29%) were positive for CD38. Both molecules were highly correlated and predictive of the clinical course of the disease. According to the simultaneous evaluation of ZAP-70 and CD38, patients were divided into three groups. In 81 patients (52%), there was a negative concordance of both molecules (ZAP-70(-)/CD38(-)); in 27 patients (17%) there was a positive concordance (ZAP-70(+)/CD38(+)); in 49 patients (31%) there was a discordant expression (ZAP-70(+)/CD38(-) and ZAP-70(-)/CD38(+)). A comparison of the clinical and laboratory data showed in ZAP-70(+)/CD38(+) patients a significantly higher bone marrow and peripheral blood lymphocytosis, lower hemoglobin levels, more advanced clinical stage, and higher number of unmutated IgV(H) status with respect to the other two groups. Furthermore, ZAP-70(+)/CD38(+) patients displayed a much shorter treatment-free interval (median 12 months vs 42 months in discordant patients and not reached in ZAP-70(-)CD38(-) patients). These results prove that the concomitant evaluation of ZAP-70 and CD38 expression allows the separation of CLL patients in prognostic subgroups and suggest that their simultaneous assessment should become an integral component of the CLL diagnostic grid. 2007 Wiley-Liss, Inc

PMID: 17534928 [PubMed - indexed for MEDLINE]

Retrospective analysis of CD38 expression in 102 patients with B-CLL with a maximum follow-up of 18 years: incidence and prognostic significance.

Medizinische Klinik II, Hämatologie und Onkologie, Klinikum Offenburg, Germany. dr.jakob@gmx.net

BACKGROUND: The aim of this study was to evaluate the incidence of CD38 expression in all CLL patients of our institution and to determine its prognostic significance in correlation with other parameters of the disease. PATIENTS AND METHODS: We analyzed the CD38 expression in 102 B-CLL patients referred to our department over a period of 12 months. RESULTS: The follow-up period ranged from 0 to 18 years. 30 patients (29%) were CD38-positive (CD38+) and 72 patients (71%) were CD38-negative (CD38-) with a median age of 65 and 64 years, respectively. Of the Binet A patients (77), 25% showed an expression of CD38; in Binet B/C patients (25), 67% expressed CD38. Median survival of the CD38- group was 77.5 months and of the CD38+ group 56.3 months. CD38 expression was associated with shorter lymphocyte doubling time (p < 0.0001), a more advanced stage of the disease, and a shorter therapy- and progression-free time (p < 0.0017/p < 0.0012), which was also true in the Binet A subgroup. In 2 cases, we detected a shift from the CD38- to the CD38+ phenotype. CONCLUSION: We found a low incidence of CD38+ CLL patients, and CD38 expression predicted significantly a more advanced stage of the disease, shorter lymphocyte doubling time and shorter therapy- and progression-free time.

PMID: 17028452 [PubMed - indexed for MEDLINE]

Comment in:

Blood. 2004 Mar 1;103(5):1968-9.

CD38 is a signaling molecule in B-cell chronic lymphocytic leukemia cells.

Laboratory of Immunogenetics, Department of Genetics, Biology and Biochemistry, via Santena 19, 10126 Turin, Italy. fabio.malavasi@unito.it

The prognosis for patients with B-cell chronic lymphocytic leukemia (B-CLL) is generally less favorable for those expressing CD38. Our working hypothesis is that CD38 is not merely a marker in B-CLL, but that it plays a receptor role with pathogenetic potential ruling the proliferation of the malignant clone. CD38 levels were generally low in the patients examined and monoclonal antibody (mAb) ligation was inefficient in signaling. Other cellular models indicated that molecular density and surface organization are critical for CD38 functionality. Interleukin 2 (IL-2) induced a marked up-modulation and surface rearrangement of CD38 in all the patients studied. On reaching a specific expression threshold, CD38 becomes an efficient receptor in purified B-CLL cells. Indeed, mAb ligation is followed by Ca2+ fluxes and by a markedly increased proliferation. The unsuitability of CD38 to perform as a receptor is obviated through close interaction with the B-cell-receptor (BCR) complex and CD19. On mAb binding, CD38 translocates to the membrane lipid microdomains, as shown by a colocalization with the GM1 ganglioside and with CD81, a raft-resident protein. Finally, CD38 signaling in IL-2-treated B-CLL cells prolonged survival and induced the appearance of plasmablasts, providing a pathogenetic hypothesis for the occurrence of Richter syndrome.

PMID: 12763926 [PubMed - indexed for MEDLINE]

Relative importance of CD38 expression over myeloid-associated markers expression in predicting the clinical course of B-CLL patients.

Department of Pathology, Medical College of Wisconsin, Milwaukee, WI 53226, USA.

Expression of CD38 or myeloid-associated markers has been reported to be important in predicting prognosis in B-cell chronic lymphocytic leukemia (B-CLL) in separate studies but the impact of combining these markers on prognosis has not been examined. The current study aimed to evaluate the relative contribution of expression of CD38 and/or myeloid-associated markers (CD11b, CD13, CD15 and CD33) by flow cytometry (FCM) on the clinical course of 24 B-CLL patients. B-CLL patients with high levels of CD38 expression, defined as greater than or equal to 30% of neoplastic lymphocytes expressing CD38, had a significantly poorer OS than those with low levels of CD38 expression (54% cumulative survival: 51 months vs. 103 months, Kaplan-Meier survival analysis, p < 0.005, Logrank test). High levels of expression of myeloid-associated markers showed no statistically significant impact on OS in these patients. Ten of 11 patients (91%) with high levels of CD38 expression required chemotherapy. In contrast, only 5 of 13 patients (38%) with low levels of CD38 expression required chemotherapy (p < 0.009, Chi Square). There was no significant difference in the requirement for chemotherapy between patients with high levels of expression of myeloid-associated marker and those without (5/8 or 63% vs. 10/16 or 63%). Thus, our results suggest that CD38 is superior to myeloid-associated markers in predicting the prognosis of patients with B-CLL. Further studies with a larger sample size are indicated to confirm our observation.

PMID: 12854897 [PubMed - indexed for MEDLINE]

Expression levels of CD38 in T cells predict course of disease in male patients with B-chronic lymphocytic leukemia.

Laboratory of Immunological and Molecular Cancer Research, 3rd Medical Dept at the Salzburg University Hospital, Muellner Hauptstrasse 48, A-5020 Salzburg, Austria. i.tinhofer@salk.at

CD38 expression of tumor cells has been identified as an important prognostic factor in B-cell chronic lymphocytic leukemia (B-CLL). Although CD38 is involved in effector functions of T cells, the prognostic value of CD38+ T cells has not yet been addressed in B-CLL. In the present study, CD38-expression levels in B-CLL cells and T cells from 204 patients were analyzed by flow cytometry and correlated with clinical and molecular risk parameters. CD38 expression significantly differed in the neoplastic clone from patients with low versus advanced stage, irrespective of the sex of patients. In contrast, CD38 expression was generally higher in T cells from female compared with male patients but only increased in male patients in a stage-dependent manner. In male patients, combined analysis of CD38 in T cells and B-CLL cells identified 4 subgroups with significantly different treatment-free survival. Multivariate analysis including Rai stage and molecular risk parameters of the neoplastic clone identified CD38-expression levels in T cells as an independent prognostic factor in male patients. Combined analysis of CD38 in B-CLL and T cells is superior in predicting outcome of male B-CLL patients than either parameter alone. Further studies are needed to elucidate the underlying mechanisms of the sex-specific role of CD38+ T cells in B-CLL.

PMID: 16825496 [PubMed - indexed for MEDLINE]

CD38 as a prognostic factor in B cell chronic lymphocytic leukaemia (B-CLL): comparison of three approaches to analyze its expression.

Department of Clinical Chemistry, University Medical Center Rotterdam, Daniel den Hoed Cancer Center, The Netherlands. j.boonstra@erasmusmc.nl

BACKGROUND: Increased CD38 expression by leukemic cells has been suggested as an adverse prognostic factor in B-CLL. Several approaches have been proposed to quantify its level of expression by flow cytometry. METHODS: We compared the use of (i) the percentage of CD38 positive cells, (ii) CD38 antibodies bound per cell (ABC), and (iii) a semi-quantitative method based on the shape of the CD38 histogram, within a cohort of 78 B-CLL patients. RESULTS: A decreased overall survival was seen with >30% CD38 positivity among B-CLL cells, with CD38 ABC >100, and with bimodal or unimodal, strongly positive CD38 histograms. However, patients with unimodal weakly positive CD38 histograms also showed a significantly reduced survival as did patients with intermediate proportions (i.e. 5-30%) of CD38+ cells. Furthermore, within the group with <5% CD38 positivity among their B-CLL cells, 84% of patients showed prognostically favourable mutated IGVH gene segments and 100% had low ZAP70 gene expression. For 5-30% CD38 positivity, these proportions were 50 and 83%, while for >30% CD38 positivity, these proportion were only 28 and 56%, respectively. CONCLUSIONS: We found a simple method of quantitation of CD38 expression (i.e., >5% CD38 positivity among B-CLL cells) to be sufficient to identify patients with an unfavourable prognosis. The level of CD38 expression as defined with this method correlated well with the IGVH mutation status and ZAP70 gene expression. Copyright 2006 International Society for Analytical Cytology.

PMID: 16568475 [PubMed - indexed for MEDLINE]

CD38 gene polymorphism and chronic lymphocytic leukemia: a role in transformation to Richter syndrome?

Department of Genetics, Biology, and Biochemistry, University of Torino Medical School, Turin, Italy.

CD38 rules proliferation signals in chronic lymphocytic leukemia (CLL) cells, suggesting that the molecule is not merely a prognostic marker but also a key element in the pathogenetic network underlying the disease. CD38 has a genetic polymorphism, characterized by a C>G variation in the regulatory region of intron 1. The working hypothesis is that the presence of different alleles in CLL patients marks (or accounts for) some of the clinical heterogeneity. CD38 allele distribution in 248 Italian patients overlapped with that of the controls (n = 232), suggesting that susceptibility to CLL is not influenced by CD38 genotype. Stratification of patients according to markers of unfavorable prognosis constantly resulted in a significantly higher frequency of the rare G allele. Furthermore, analysis of clinical parameters showed that G allele is independently associated with nodal/splenic involvement. The highest G allele frequency was observed in the 16 patients of the cohort that developed Richter syndrome (RS). Five-year cumulative incidence of transformation was significantly higher in G allele carriers than in CC homozygotes. Multivariate analysis on a total of 30 RS patients confirmed that the probability of transformation is strongly associated with G allele, likely representing an independent risk factor for RS development.

PMID: 18424664 [PubMed - indexed for MEDLINE]

CD38 expression in chronic lymphocytic leukemia is regulated by the tumor microenvironment.

Department of Haematological and Molecular Medicine, King's College London, Rayne Institute, London, USA. piers.patten@kcl.ac.uk

Chronic lymphocytic leukemia (CLL) is a lymphoproliferative disease with a highly variable outcome. The prognosis of patients with CLL may be predicted using a number of biomarkers, including the level of CD38 expression at the leukemic cell surface. This study investigates the hypothesis that CD38 expression by CLL cells reflects interactions with nonmalignant cells within pseudofollicles in secondary lymphoid tissue where tumor cell proliferation is thought to occur. CD38 expression is higher in tissues that contain pseudofollicles compared with those that do not. In addition, we show that CD38 expression in CLL is dynamic, changes in response to contact with activated CD4(+) T cells, and identifies cells that are primed to proliferate. Finally, we demonstrate close contact between activated CD4(+) T cells and proliferating tumor in primary patient tissue. Proliferating tumor cells in lymph nodes express CD38, which is in turn associated with an increased number of CD31(+) vascular endothelial cells. Although the factors resulting in colocalization of tumor, T cells, and endothelium remain unclear, the existence of these cellular clusters may provide an explanation for the association between CD38 expression and adverse outcome in CLL and suggests novel therapeutic targets.

PMID: 18326821 [PubMed - indexed for MEDLINE]

No convincing evidence for a role of CD31-CD38 interactions in the pathogenesis of chronic lymphocytic leukemia.

Department of Hematology, Academic Medical Center, Amsterdam, The Netherlands. s.h.tonino@amc.uva.nl

Although CD38, a marker of poor prognosis in chronic lymphocytic leukemia (CLL), is known primarily as an ecto-enzyme, it has also been ascribed a receptor function. Interaction with its proposed ligand CD31 expressed on nurse-like cells would result in proliferative and survival-signals. Yet, in CLL, both homotypic and heterotypic CD31-CD38 interactions are expected to be rather ubiquitous. We analyzed whether CD38-CD31 interactions result in proliferative and antiapoptotic signals. We found a high expression of CD31 on CLL, irrespective of CD38 expression. Coculture of CD38(high) CLL with endothelial cells or CD31 transfected fibroblasts, with or without blocking CD31 or CD38 antibodies, did not result in increased survival or proliferation. Analysis of gene expression of most known regulators of apoptosis revealed no influence of coculture with CD31-expressing feeder cells. In conclusion, our data do not support an important contribution of CD38 triggering by CD31 to the proliferative and antiapoptotic state of the leukemic clone.

PMID: 18519815 [PubMed - indexed for MEDLINE]

Gene expression signatures separate B-cell chronic lymphocytic leukaemia prognostic subgroups defined by ZAP-70 and CD38 expression status.

Clinic of Hematology, University Hospital, University of Duisburg-Essen, Essen, Germany.

B-cell chronic lymphocytic leukaemia (B-CLL) is a heterogenous disease with a highly variable clinical course and analysis of zeta-associated protein 70 (ZAP-70) and CD38 expression on B-CLL cells allowed for identification of patients with good (ZAP-70-CD38-) and poor (ZAP-70+CD38+) prognosis. DNA microarray technology was employed to compare eight ZAP-70+CD38+ with eight ZAP-70-CD38- B-CLL cases. The expression of 358 genes differed significantly between the two subgroups, including genes involved in B-cell receptor signaling, angiogenesis and lymphomagenesis. Three of these genes, that is, immune receptor translocation-associated protein 4 (IRTA4)/Fc receptor homologue 2 (FcRH2), angiopoietin 2 (ANGPT2) and Pim2 were selected for further validating studies in a cohort of 94 B-CLL patients. IRTA4/FcRH2 expression as detected by flow cytometry was significantly lower in the poor prognosis subgroup as compared to ZAP-70-CD38- B-CLL cells. In healthy individuals, IRTA4/FcRH2 protein expression was associated with a CD19+CD27+ memory cell phenotype. ANGPT2 plasma concentrations were twofold higher in the poor prognosis subgroup (P<0.05). Pim2 was significantly overexpressed in poor prognosis cases and Binet stage C. Disease progression may be related to proangiogenic processes and strong Pim2 expression.

PMID: 16932341 [PubMed - indexed for MEDLINE]