T type 1/type 2 subsets balance in B-cell chronic lymphocytic leukemia—the three-color flow cytometry analysis

2002 ◽  
Vol 26 (7) ◽  
pp. 657-660 ◽  
Author(s):  
Monika Podhorecka ◽  
Anna Dmoszynska ◽  
Jacek Rolinski ◽  
Ewa Wasik
Keyword(s):  
Flow Cytometry ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Lymphocytic Leukemia ◽  
Flow Cytometry Analysis ◽  
Color Flow ◽  
Cell Chronic Lymphocytic Leukemia

Detection of t(14;18)(q32;q21) in B-Cell Chronic Lymphocytic Leukemia

2005 ◽  
Vol 129 (3) ◽  
pp. 410-411
Author(s):  
Wolfgang Kern ◽  
Torsten Haferlach ◽  
Susanne Schnittger ◽  
Claudia Schoch
Keyword(s):  
Flow Cytometry ◽  
In Situ Hybridization ◽  
Chronic Lymphocytic Leukemia ◽  
Follicular Lymphoma ◽  
B Cell ◽  
Lymphocytic Leukemia ◽  
Multiparameter Flow Cytometry ◽  
Lymphoma Therapy ◽  
Cell Chronic Lymphocytic Leukemia

Abstract Cytomorphologic testing and multiparameter flow cytometry are the mainstays in diagnosing B-cell chronic lymphocytic leukemia, whereas fluorescence in situ hybridization that targets the translocation t(14;18)(q32;q21) often is used to identify follicular lymphoma. Therapy is highly diverse between both diseases. We describe a case with cytomorphologically and immunologically proven B-cell chronic lymphocytic leukemia in which t(14;18)(q32;q21) was found.

CD38 expression as an important prognostic factor in B-cell chronic lymphocytic leukemia

Blood ◽  
2001 ◽  
Vol 98 (1) ◽  
pp. 181-186 ◽  
Author(s):  
Sherif Ibrahim ◽  
Michael Keating ◽  
Kim-Anh Do ◽  
Susan O'Brien ◽  
Yang O. Huh ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Prognostic Factor ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Lymphocytic Leukemia ◽  
Leukemic Cells ◽  
Aggressive Disease ◽  
Cd38 Expression ◽  
Important Prognostic Factor ◽  
Cell Chronic Lymphocytic Leukemia

Abstract CD38 is a transmembrane glycoprotein expressed on the surface of leukemic cells in a significant percentage of patients with B-cell chronic lymphocytic leukemia (B-CLL). A recent study suggested that CD38 expression has prognostic value in CLL. Peripheral blood samples from 218 patients with B-CLL were analyzed by flow cytometry for CD38 expression on CD5/19+ leukemic cells. Various patient characteristics were studied including age, sex, Rai and Binet stages, splenomegaly, hepatomegaly, hemoglobin (Hgb) level, β-2 microglobulin (β2M) level in the serum, number of nodal sites involved with disease, and length of survival. The Kaplan-Meier method was used to construct survival curves, and the log-rank statistic was used to compare these curves. CD38 was expressed in 20% or more of leukemic cells in 43% of the patients. Patients with high CD38 expression (20% or more) had significantly shorter survival times (P =.00005). Multivariate analyses showed that CD38 expression is an important prognostic factor associated with high incidence of lymph node involvement (P = .004), lower hemoglobin level (P = .001), hepatomegaly (P = .05), and high β2M level (P = .00005). CD38 expression identified a group of patients with aggressive disease that was considered by Rai staging to be early-stage disease (Rai stages 0-II). Patients with CD38+ samples have significantly aggressive disease regardless of their clinical stage. Measurement of CD38 expression by flow cytometry should become a routine test in the evaluation of patients with CLL.

Monoclonal B-Cell Lymphocytosis Is Commonly Observed Among Unaffected Members of High Risk CLL Families.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1232-1232 ◽  
Author(s):  
Lynn R Goldin ◽  
Mark C Lanasa ◽  
Susan L Slager ◽  
Sara S Strom ◽  
Nicola J Camp ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Risk ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Germ Line ◽  
Lymphocytic Leukemia ◽  
Detection Sensitivity ◽  
Flow Cytometry Analysis ◽  
First Degree Relatives ◽  
High Risk Families

Abstract Abstract 1232 Poster Board I-254 B-Chronic Lymphocytic Leukemia (CLL) shows a strong familial risk and also co-aggregates with other indolent lymphomas. Monoclonal B-cell lymphocytosis (MBL) is an asymptomatic hematologic condition characterized by small B-cell clones, most with a surface phenotype similar to that of chronic lymphocytic leukemia (CLL). These clones are detectable at low absolute lymphocyte cell (ALC) numbers in otherwise healthy individuals using sensitive 6 or 8 color flow cytometry analysis and can be a precursor to CLL. In the general population, MBL increases with age with a prevalence of 5% in individuals over age 65. In contrast, the rate of MBL is 13-18% in first degree relatives of CLL patients in high risk families. In the largest study to date, we report the characteristics of MBL among 430 first-degree relatives of CLL patients (with no associated lymphoproliferative diseases [LPD]) in 132 high risk families. Patients and Methods Individuals studied came from “high risk” families (defined as those with two or more confirmed cases of CLL) that are participating in the Genetic Epidemiology of CLL Consortium. Multi-parameter flow cytometry analysis with a detection sensitivity of 0.02% was performed on either fresh or cryopreserved PBMC and MBL was classified according to previously published criteria (Marti et al, Br. J Haematol 130:325, 2005). Both the ALC and the absolute B cell count (B-ALC) were calculated for each individual. Survival analysis was used to compute the probability of developing MBL with age using the life table method. Results The overall rate of MBL was 17% (73/430) among first–degree relatives but the probability for developing MBL by age 90 was 51%. Males had a slightly higher (but non-significant) risk for MBL than females. MBL patients had significantly higher ALC and B-ALC than did those with normal immunophenotype. The mean ALC was 2.5×109/L among MBL patients, and 20% had an ALC greater than 3 ×109/L. The B-ALC count averaged 0.51 ×109/L. Ninety percent of the MBL cases had a CLL-like phenotype (CD20dim, CD5+, CD23+). Conclusions MBL is found at a very high rate in families selected for having 2 or more patients with CLL suggesting that MBL reflects inherited predisposition to CLL. Although most of the MBL cases in these families had low cell counts and thus have a presumed low likelihood of progressing to CLL or other LPD, a higher proportion of them had lymphocytosis compared to those with normal immunophenotype. We hypothesize that if MBL is an early step in the process of development of CLL, then germ line genes are likely to be acting early in carcinogenesis with more “hits” required before CLL develops. By looking for genes associated with MBL or CLL in families or in the population, we could substantially increase our power to identify germ line genes predisposing to CLL. Disclosures Kay: Biogenc-Idec, Celgene, Genentech, genmab: Membership on an entity's Board of Directors or advisory committees; Genentech, Celgene, Hospira, Polyphenon Pharma, Sanofi-Aventis: Research Funding.

The Earliest Activation Marker CD69 Is An Independent and Strong Progression Indicator in B-Cell Chronic Lymphocytic Leukemia.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2359-2359
Author(s):  
Giovanni Del Poeta ◽  
Maria Ilaria Del Principe ◽  
Cristina Simotti ◽  
Francesco Buccisano ◽  
Luca Maurillo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Multivariate Analysis ◽  
Prognostic Factor ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Conflicts Of Interest ◽  
Lymphocytic Leukemia ◽  
Significant Prognostic Factor ◽  
Therapeutic Decisions ◽  
Cell Chronic Lymphocytic Leukemia

Abstract Abstract 2359 Poster Board II-336 B-cell chronic lymphocytic leukemia (B-CLL) exhibits features of activated and antigen-experienced B-lymphocytes and CD69 overexpression resembles B cells at an earlier and greater state of activation (Damle, 2002 and 2007). Moreover, the recent in vitro generation of anti-CD69 monoclonal antibodies able to inhibit either tumor growth or enhance NK cell function (Esplugues, 2005) prompt us to extensively evaluate CD69 antigen in our B-CLL cases. The primary endpoints of our research were: 1) to determine progression-free survival (PFS) and overall survival (OS) upon CD69; 2) to assess the additive prognostic value of CD69 and ZAP-70 and finally 3) to confirm CD69 as an independent prognostic factor. We investigated 401 patients (pts), median age 65 years, 219 males and 182 females. With regard to modified Rai stages, 120 pts had a low stage, 263 an intermediate stage and 18 a high stage. ZAP-70 and CD69 were determined by multicolor flow cytometry, fixing the cut-off values > 20% and >30%, respectively. ZAP-70+ and CD69+ pts were 166/401 (41%) and 108/401 (27%), respectively. CD69 lower than 30% was significantly associated with low Rai stage (105/120; P<0.0001), lymphocyte doubling time >12 months (258/332; P=0.00001), beta-2 microglobulin (B-2M) <2.2 mg/dl (181/223; P=0.00004) and soluble CD23 <70 U/ml (196/240; P<0.00001). There were significant correlations between lower CD69 and IgVH gene mutated status (283 total cases, 150/210; P=0.0001) or low risk (normal or 13q-) FISH cytogenetics (296 total cases, 149/213; P<0.00001). Equally, a strict association was found between lower CD69 and lower ZAP-70 (185/293; P=0.0003). With regard to clinical outcome, both a shorter PFS and OS were observed in ZAP-70+ pts (6% vs 56% at 12 years and 30% vs 95% at 16 years; P<0.00001) as well as in CD69+ pts (9% vs 49% at 14 years, P<0.00001 and 49% vs 75% at 16 years; P=0.00002). Noteworthy, ZAP-70 and CD69 showed additive prognostic properties, since ZAP-70 <20% plus CD69 <30% identified a B-CLL subset at better prognosis with regard to PFS (65% vs 2% at 12 years; P<0.00001, Figure) and OS (97% vs 35% at 14 years; P<0.00001). The two discordant subsets (CD69+ZAP-70 negative and CD69-ZAP-70+) showed an intermediate outcome (Figure). In multivariate analysis of PFS, CD69 (P=0.001) and ZAP-70 (P=0.005) together with cytogenetics and B-2M were confirmed to be independent prognostic factors. On the other hand, with regard to multivariate analysis of OS, age > or < 60 years resulted to be the most significant prognostic factor (P=0.004) followed by CD38 (P=0.01), IgVH status (P=0.02) and ZAP-70 (P=0.04). In conclusion, CD69 antigen, determined by flow cytometry, should be considered a novel important prognostic parameter in B-CLL and its easy and rapid laboratory determination may allow us to identify early progressive pts in order to take timely therapeutic decisions. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Detection of CD5 in B-cell chronic lymphoproliferative diseases by flow cytometry: a strong expression in B-cell chronic lymphocytic leukemia

2005 ◽  
Vol 20 (suppl 1) ◽  
pp. 56-62
Author(s):  
Geraldo Barroso Cavalcanti Júnior ◽  
Valeria Soraya de Farias Sales ◽  
Dany Geraldo Kramer Cavalcanti e Silva ◽  
Maria Cleide de Araújo Lopes ◽  
Aldair de Souza Paiva ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Lymphoproliferative Disease ◽  
Lymphocytic Leukemia ◽  
Leukemic Cells ◽  
B Cell Malignancies ◽  
Prolymphocytic Leukemia ◽  
Chronic Lymphoproliferative Diseases ◽  
Cell Chronic Lymphocytic Leukemia

PURPOSE: CD5 is a T cell marker, aberrantly express in B cell chronic lymphocytic leukemia (B-CLL) and mantle cell lymphoma (MCL). Other chronic B cell malignancies including hairy cell leukemia (HCL) and B cell prolymphocytic leukemia (B-PLL) are CD5 negative or express this antigen in a weak way. In this study, CD5 expression was investigated in leukemic cells from 42 patients with chronic B cell lymphoproliferative disease. METHODS: We studied the CD5 expression in leukemic cells from 42 patients with chronic B-cell malignancies by flow cytometry. Demographic features such as age, sex and clinical date were also analyzed. RESULTS: There were 22 males and 20 females. The immunophenotyping showed that 35 cases were B-CLL, 3 B-PLL and HCL and one patient was MCL. CD5 expression was present in all B-CLL and MCL. Low expression of CD5 was observed in one patient with B-PLL and negative in all cases of HCL. CONCLUSION: Our date demonstrated that CD5 expression can help distinguish among B-CLL from HCL and B-PLL, but is similar expressed in MCL.

scholarly journals Expression of FAK and Its Involvement in the Progression of B-Cell Chronic Lymphocytic Leukemia

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3309-3309
Author(s):  
Cristina Gattazzo ◽  
Andrea Visentin ◽  
Alberto Pavan ◽  
Veronica Martini ◽  
Federica Frezzato ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
B Cells ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
B Lymphocytes ◽  
Focal Adhesion ◽  
Poor Prognosis ◽  
Lymphocytic Leukemia ◽  
Bcr Signaling ◽  
Cell Chronic Lymphocytic Leukemia

Abstract INTRODUCTION B-cell chronic lymphocytic leukemia (B-CLL) is a disorder characterized by the accumulation of clonal CD5+ B lymphocytes, due to uncontrolled growth and resistance to apoptosis. Although the prognosis and clinical outcome has dramatically improved by recent innovative therapies, B-CLL still remains an incurable disease. Since signaling events downstream the BCR engagement are important for the progression of B cells, BCR signaling has been investigated in B-CLL in order to design new agents to specifically treat this disease. We demonstrated that Lyn, one of the first kinases involved in BCR signaling pathway, is overexpressed, constitutively active and anomalously distributed in malignant B cells, as compared to normal B lymphocytes. The Focal adhesion kinase (FAK), a non-receptor protein tyrosine kinase, is the primary enzyme involved in the engagement of integrins and assembly of Focal Adhesion. FAK is regulated primarily through tyrosine phosphorylation by Lyn after BCR engagement and was found to be overexpressed in many kinds of human cancers. However, a downmodulation of FAK expression and its association to poor prognosis have also been reported. The aim of this study was to investigate the role of FAK in CLL patients. METHODS Blood samples were collected from 5 controls and 50 B-CLL patients. Informed consent was obtained according to the Declaration of Helsinki. Untouched peripheral blood B cells were purified using the RosetteSep for human B cells isolation kit. The samples that were used had at least 95% of normal CD19+ or neoplastic CD5+/CD19+ cells, as assessed by flow-cytometry. Level of FAK protein was evaluated by Western blotting (Wb) and Flow Cytometry assay (FC). Levels of FAK were correlated to clinical parameters of patients. RESULTS We observed that FAK was downmodulated in 56% of analyzed patients with respect to healthy subjects (respectively, Wb: 0.28±0.25 vs 0.85±0.32, p<0.001; FC: 35%±29 vs 60%±16, p<0.05). We also identified that lower levels of FAK expression were related to the prognostic markers of poor outcome (the expression of ZAP70, CD38 and an unmutated-IGHV genes status, p<0.05) and to a shorter Treatment Free Survival (p<0.05). Moreover, patients (n=6) who had an indolent course and were responsive to the standard treatment, showed normal expression of this kinase already at diagnosis. In contrast, patients (n=6) with a more aggressive disease, had a lower expression of FAK, that was further downmodulated during the progression of disease, irrespective of how the patients were treated. CONCLUSIONS From the data presented in this report we propose that FAK downmodulation could be considered as a new marker of poor prognosis and as a putative predictor for high-risk subgroups of CLL, even in early-stage disease. Disclosures No relevant conflicts of interest to declare.

scholarly journals Neurotensin receptor type 2 protects B-cell chronic lymphocytic leukemia cells from apoptosis

Oncogene ◽  
2017 ◽  
Vol 37 (6) ◽  
pp. 756-767 ◽  
Author(s):  
A Abbaci ◽  
H Talbot ◽  
S Saada ◽  
N Gachard ◽  
J Abraham ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Lymphocytic Leukemia ◽  
Receptor Type ◽  
Leukemia Cells ◽  
Neurotensin Receptor ◽  
Cell Chronic Lymphocytic Leukemia

Combined Schedule of Fludarabine and Alemtuzumab Followed by Maintenance with Low-Dose Alemtuzumab Is Effective for Patients with Progressive B-Cell Chronic Lymphocytic Leukemia.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5048-5048
Author(s):  
Malgorzata Kowal ◽  
Wieslaw Nowak ◽  
Aleksandra Nowaczynska ◽  
Anna Dmoszynska ◽  
Aleksander B. Skotnicki
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Residual Disease ◽  
Therapeutic Option ◽  
Single Agent ◽  
Elevated Serum ◽  
Lymphocytic Leukemia ◽  
Color Flow ◽  
Cell Chronic Lymphocytic Leukemia

Abstract Patients with relapsed/refractory B-cell chronic lymphocytic leukemia (B-CLL) have a poor treatment outcome. Most patients who are resistant to single agent fludarabine or alemtuzumab (Campath®; anti-CD52 monoclonal antibody) do not survive longer than 9 months; however, combination fludarabine and alemtuzumab (FluCam) appears to have an additive effect in patients who are resistant to either agent alone. This study assessed the efficacy and safety of alemtuzumab at a reduced dose, first in combination with fludarabine and then as monotherapy in patients with relapsed or refractory B-CLL. Patients received fludarabine 25 mg/m2 IV over 30 minutes, and alemtuzumab 10 mg IV over 2 hours for 3 days. Alemtuzumab was titrated for the first 1–3 days and administered with prophylactic antihistamine and antipyretic agents. Acyclovir and cotrimoxazole were used to prevent infection and manage inflammatory complications. Treatment cycles were repeated every 4 weeks for up to 6 cycles, and following successful completion of FluCam, alemtuzumab was administered monthly for 4–5 doses. Response and toxicity were evaluated according to the current criteria of NCI-WG and WHO, respectively. Minimal residual disease (MRD) was assessed with four-color flow cytometry. A total of 10 patients, 33–57 years of age (mean, 48 years), with relapsed or refractory B-CLL received FluCam. Mean time from diagnosis was 6 (range, 2–8) years. Patients had disease classified as Rai stages IV (n=6), III (n=1), II (n=1), and I (n=2). All patients had elevated serum LDH and beta-2 microglobulin, and increased ZAP-70 expression. Patients received a mean of 3 (range, 2-6) prior courses of chemotherapy; 1 patient previously received alemtuzumab, and 6 other patients previously received combination fludarabine and cyclophosphamide. Patients received an average of 4 cycles of FluCam for a total of 37 courses. Following combination FluCam, 3 patients received alemtuzumab on average for 3 (range, 2–5) months. Mean duration of follow-up was 11 (range, 3–27) months. Complete remission was achieved in 2 patients following 3 cycles of FluCam; 1 patient achieved MRD negativity. Short term improvements were seen in 2 patients; however, treatment was discontinued after 2 and 3 cycles because of disease progression. Partial remission was obtained in 6 patients, including total regression of general symptoms and 75% regression of lymphadenopathy and splenomegaly. Moderate side effects including chills, fever, and skin redness were observed in all patients during alemtuzumab dose escalation. Inflammatory complications of grades 2/3 occurred in 1 patient following the first 2 courses of FluCam. One patient with severe immune deficiency died after 27 months of follow up due to bacterial pneumonia. CMV reactivation did not occur in any patient. Results from this study suggest that combination FluCam is active in patients with advanced stages of B-CLL, and may provide a therapeutic option for patients who are refractory to either agent alone. Maintenance therapy with alemtuzumab may prolong time to progression and overall survival without increasing toxicity or hematologic and inflammatory complications in patients with relapsed or refractory B-CLL. Further study is warranted in a larger patient population to confirm these results.

scholarly journals The Diagnostic Value of Flow Cytometry Imunophenotyping in an Albanian Patient Population with a Preliminary Clinical Diagnosis of Chronic Lymphocytic Leukemia

2014 ◽  
Vol 2 (1) ◽  
pp. 51-55
Author(s):  
Valentina Semanaj ◽  
Arbi Pecani ◽  
Teuta Dedej ◽  
Alma Barbullushi ◽  
Zamira Ylli ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Cell Population ◽  
Clinical Stage ◽  
Final Diagnosis ◽  
Diagnostic Value ◽  
Lymphocytic Leukemia ◽  
Advanced Clinical Stage ◽  
Color Flow

Objective: Based on the flow cytometry multiparametric immunophenotyping methodology we studied some useful cell marker criteria needed for the practical differentiation of the chronic lymphocytic leukemia from other chronic limphoproliferative diseases with a leukemic component.Materials and Methods: The applied methodology is a four color flow cytometry multiparametric immunophenotyping technique using EDTA blood samples taken from 84 consecutive patients diagnosed with CLL through a preliminary clinical and white blood cell examination. The following fluorescent stained monoclonal antibodies were used: CD3, CD4, CD5, CD8, CD11c, CD19, CD20, CD23, CD25, FMC7 and kappa/lambda light chains.Results: From the 84 individuals tested, 2 out of them (2.4%) resulted with a abnormal T-cell population while 82 (97.6%) showed a pathological B cell line. 58 (69.1%) patients resulted with typical CLL markers (CD19+CD5+CD23+) while 5 (5.9%) of them presented a non typical chronic lymphocytic leukemia profile (CD19+CD5+CD23-). 19 (22.6%) out of patients displayed an abnormal CD19+CD5- B cell population. A statistically significant correlation was found between the clinical stage of CLL and the positivity for the CD38 marker (p=0.04).Conclusion: Flow cytometry immunophenotyping is a fundamental examination for the final diagnosis of chronic lymphocytic leukemia. The expression of CD38+ in CLL patients stands for a more advanced clinical stage.

Sign in / Sign up

Export Citation Format

Share Document