scholarly journals Phenotypic Characteristics of Tumor Cells in Patients with Chronic Lymphocytic Leukemia into Different Prognostic Groups

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 5276-5276
Author(s):  
Aleksei Kuvshinov ◽  
Sergei Voloshin ◽  
Irina Martynkevich ◽  
Ludmila Martynenko ◽  
Andrei Garifullin ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Tumor Cells ◽  
Conflicts Of Interest ◽  
Lymphocytic Leukemia ◽  
Phenotypic Characteristics ◽  
Hla Dr ◽  
Number Of Patients ◽  
Genetic Abnormalities ◽  
Prognostic Groups ◽  
The Relationship

Abstract Background. The presence or absence of certain cluster of differentiation on the tumor cells of chronic lymphocytic leukemia may affect the course of the disease. Influence of genetic abnormalities on the prognosis of the disease was also proved. Aim. To determine the relationship of the phenotype of tumor cells with genetic prognostic groups of patients with chronic lymphocytic leukemia (CLL). Methods. Thirty-five adult pts (median age 61 year, range 44 - 82; male 24, female 11) with diagnosed CLL were included in the study. The CLL was diagnosed according to the standard basic examination (complete blood count with differential, multicolor flow cytometry (MFC) of blood and bone marrow (BM), lymph node and BM immunohistochemistry (IHC), computered tomography). Cytogenetic studies were performed on blood samples using standard GTG-method. Interphase FISH analyses were performed according to the manufacturer's protocol using DNA probes: LSI 13(RB1)13q14, LSI ATM (11q22), CEP12, LSI TP53 (17p13.1) (Abbott). Immunophenotype (IFT) of CLL cells assessed with combinations: CD3/CD19, CD19/CD5, CD19/CD11c, CD19/CD20, CD19/CD22, CD19/CD23, CD19/CD25, CD19/CD38, CD19/CD43, CD19/CD81, CD19/HLA-DR, and CD19/CD5/CD23. Results. Stratification of patients into prognostic groups was performed based on identified GA. Favorable prognosis - patients with del(13q) (n = 9); neutral prognosis - normal karyotype (n = 14) or trisomy of chromosome 12 (n = 4); unfavorable prognosis - del(17p) (n = 3), del(11q) (n = 3) and the complex karyotype (n = 2). Expression of CD20 was lower, and CD38 - higher in adverse group (51.0±16.31 % and 36.02±10.35 %, respectively) versus neutral or favorable groups (CD20+ - 83.17±5.52 % and 84.41±4.7 %; CD38 - 10.46±4.8 %, and 12.44±4.1 %, respectively, p <0.05). The expression level of CD20 and CD38 did not differ between the neutral and favorable groups. The number of patients with CD38 expression more than 10% was higher in the unfavorable group (7/8) versus favorable (4/8) (p <0.05). At the same time overexpression of CD38 was observed more frequently in patients with a lack of expression of CD23 on CD5 tumor cells (CD5+CD23-) (p<0.05). Expression of HLA-DR was higher in patients with MRD-negative remissions (3/4) versus patients with MRD-positive remissions (1/8) (p<0.05). Conclusions. The study of the influence of various factors on the prognosis and course of CLL requires a comprehensive approach. Further researches are needed to determine the relationship between CLL affecting factors like genetic abnormalities, phenotypic characteristics of the tumor cells and MRD. Disclosures No relevant conflicts of interest to declare.

The Composition of the B Cell Receptor Repertoire In 7428 Cases of Chronic Lymphocytic Leukemia: One Third Stereotyped, Two Thirds Heterogeneous - What Does This Mean?

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 43-43 ◽  
Author(s):  
Andreas Agathaggelidis ◽  
Nikos Darzentas ◽  
Anastasia Hadzidimitriou ◽  
Xavier Brochet ◽  
Fiona Murray ◽  
...  
Keyword(s):  
Amino Acid ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Conflicts Of Interest ◽  
Critical Mass ◽  
Relative Size ◽  
Ground Level ◽  
Lymphocytic Leukemia ◽  
Cluster Assignment ◽  
Number Of Patients

Abstract Abstract 43 Chronic lymphocytic leukemia (CLL) is characterized by the existence of subsets (clusters) of cases with restricted, “stereotyped” immunoglobulin (IG) variable heavy complementarity-determining region 3 (VH CDR3) sequences within their B cell receptors (BcR), suggesting selection by common epitopes or classes of structurally similar epitopes. Emerging evidence indicates that the grouping of CLL cases into distinct clusters with “stereotyped” BcR is functionally and prognostically relevant. Further than that, several issues remain open: (i) the refinement of criteria for identification of BcR stereotypy and cluster assignment; (ii) the true frequency of BcR stereotypy; (iii) the total number of clusters and relative size of each; and, (iv) the identification of “CLL-biased” features in BcR stereotypes. To address these issues, we systematically examined VH CDR3 stereotypy in 7596 IGHV-D-J sequences from 7428 patients with CLL (168 cases, 2.2%, with two productive sequences), three times the size of the largest published series. Recent studies in both normal B cells and other (non-CLL) B cell malignancies along with accumulated experience in our group led to an advanced clustering bioinformatics algorithm applying more stringent criteria than before. A novel parameter was also included; the usage of IGHV genes, which takes into account the role of the germline-encoded specificities in (super)antigen recognition. The algorithm assigns sequences in a cluster only if exhibiting >50% amino acid identity and >70% amino acid VH CDR3 similarity and also carrying IGHV genes that share common ancestry and, thus, belong to the same IGHV phylogenetic clan. To increase the likelihood that cluster assignment reflects actual structural relatedness, we also required that each cluster consisted only of sequences with identical VH CDR3 length and identical offsets of common patterns. Following this new approach, 2308/7596 (30.4%) CLL sequences were assigned to 952 different ground-level clusters with shared patterns and unique characteristics, each containing 2 to 56 cases. Different types of VH CDR3 patterns were identified, independent of mutational status, as “mainly germline”, i.e. deriving from restricted associations of specific IGHD and IGHJ genes, and “junctional+germline”, i.e. extending over V-D and/or D-J junctions as well. In several clusters of mutated sequences, the cluster-defining features were ubiquitous junctional residues. Common sequences among ground-level clusters enabled grouping into clearly delineated, higher-order (HO) clusters that were considerably larger in size and displayed ‘CLL-biased’ features with regard to: IGHV gene usage, somatic hypermutation (for clusters with mutated sequences) and VH CDR3 pattern composition. As an example, the largest HO cluster, including 213 sequences (2.8% of the cohort), utilized the IGHV3-21 gene with an acidic residue at VH CDR3 position 107 (3 of 9), while the second-ranking HO cluster, including 184 sequences (2.1% of the cohort), utilized different IGHV genes of Clan I (e.g. IGHV1-2, 1–3, 1–8, 1–18, 5-a, 7-4-1) with a QWL motif at VH CDR3 positions 108–110 (4-6 of 13). Based on random set simulations (using the actual sequences) and starting from a critical mass of 2000 cases, each increase of the total set by a 1000 random cases resulted in an increase in the percentage of stereotypy to ∼2% (i.e. from 21% in 2000 cases to 25% in 3000 cases to 30% in 7000), though not proportional to the increase of the cohort. Perhaps most important, however, was the finding that the percentage of sequences in known major clusters was remarkably stable compared to previous studies on smaller series. These results strongly indicate that not all CLL belong to stereotyped subsets even if the cohort size is increased significantly, corroborating our previous hypothesis that CLL consists of two distinct categories, one with stereotyped and the other with heterogeneous BcR, likely of different ontogenetic origin. Furthermore, they demonstrate that the major clusters collectively represent a sizeable proportion of the cohort. Consequently, this deeper, more robust, compartmentalized examination of BcR structures in association with other biological and clinical information may eventually pave the way for the introduction of specialized treatment protocols applicable to a significant number of patients assigned to the same cluster. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Suppression of CXCL-1 Could Restore Necroptotic Pathway in Chronic Lymphocytic Leukemia

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5466-5466
Author(s):  
Zhao Xu ◽  
Yifeng Sun ◽  
Zheng Wei ◽  
Peng Liu
Keyword(s):  
Flow Cytometry ◽  
Chronic Lymphocytic Leukemia ◽  
Western Blot ◽  
Conflicts Of Interest ◽  
Lymphocytic Leukemia ◽  
Rt Pcr ◽  
Gm Csf ◽  
Binding Factor ◽  
Tnf Α ◽  
The Relationship

The defective necroptotic pathway and cytokines were important in the pathogenesis and progression of chronic lymphocytic leukemia (CLL). On the other hand, selenite could induce necroptosis in various sold malignancies by generating reactive oxygen species. However, the relationship between necroptosis and cytokines still remains unclear. Here, we managed to clarify the role of different cytokines and selenite in the defective necroptotic pathway of CLL. We first screened the expression of different cytokines related to malignancies between CLL patients and controls using Realtime RT-PCR. Only the expression of CXCL-1, MCP-1, IL-6 and GM-CSF was significantly higher in CLL patients than that of controls. (Figure A, B) In normal B lymphocytes, TNF-α and z-VAD could induce necroptosis and also downregulated the expression of CXCL-1 and MCP-1, which indicated that CXCL-1 and MCP-1 might have correlation with necroptosis. (Figure C) After knockdown of CXCL-1 rather than MCP-1 by siRNA, CLL cells restored the TNF-α/z-VAD induced necroptosis measured by flow cytometry. (Figure D, E) To assess the association between CXCL-1 and lymphoid enhancer-binding factor 1 (LEF-1), which has already been confirmed as the key protein in the necroptotic pathway of CLL, we first performed flow cytometry to verify that CLL cells restored TNF-α/z-VAD induced necroptosis after inhibition of either CXCL-1 or LEF-1. (Figure F) Then, we used Realtime RT-PCR, Western Blot and ELISA to confirm that the expression of LEF-1 was downregulated after inhibiting the expression of CXCL-1 by siRNA, however, the expression of CXCL-1 did not change significantly after knockdown of LEF-1. (Figure G, H, I) This results demonstrated that CXCL-1 located the upstream of LEF-1. To figure out the relationship between cytokines, necroptosis and selenite, we first construct the selenite concentration gradient and selenite with different concentrations was added into CLL cells together with TNF-α and z-VAD. We found that selenite could downregulate the expression of CXCL-1 but had little influence on LEF-1 measured by Realtime RT-PCR. (Figure J) Then, flow cytometry was performed to calculate the percentage of survival CLL cells and only 3.2μM selenite could significantly induce necroptosis of CLL cells. (p = 0.0102, Figure K) Finally, both Western Blot and ELISA showed the similar result that 3.2μM sodium selenite downregulated the translational expression of CXCL-1 but had little impact on LEF-1. (Figure L) Figure Disclosures No relevant conflicts of interest to declare.

Arsenic Trioxide Induces Apoptosis In Chronic Lymphocytic Leukemic Cells Through Down-Regulation Of Survivin Via The p53-Dependent Signaling Pathway

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 5313-5313
Author(s):  
Xiaohui Zhang ◽  
Meng Lv ◽  
Qian Jiang ◽  
Honghu Zhu ◽  
Yazhen Qin ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Arsenic Trioxide ◽  
Signaling Pathway ◽  
Tumor Cells ◽  
Conflicts Of Interest ◽  
Lymphocytic Leukemia ◽  
Dependent Manner ◽  
Down Regulation ◽  
Knock Down ◽  
Dependent Signaling Pathway

Abstract Arsenic trioxide induces cell apoptosis in a variety of tumor cells. However, the molecular mechanism underlying the killing effects of arsenic trioxide on tumor cells is not fully understood. Here, we demonstrate a signal pathway of arsenic trioxide treatment that leads to apoptosis in a B-cell chronic lymphocytic leukemia (WSU-CLL) cell line. Arsenic trioxide treatment significantly inhibited proliferation and induced apoptosis of WSU-CLL cells in a time- and dose-dependent manner. This anti-tumor effect of arsenic trioxide on WSU-CLL cells was mediated through the down-regulation of survivin and up-regulation of p53. Knock-down of intracellular survivin with siRNA potentiated the inhibitory effect of arsenic trioxide on proliferation in WSU-CLL cells. Also, knock-down of p53 by a specific siRNA prevented the down-regulation of survivin by arsenic trioxide and reduced the cytotoxic effect of arsenic trioxide to WSU-CLL cells. These results indicate that arsenic trioxide may be used to as novel therapeutic agent for chronic lymphocytic leukemia, which down-regulates survivin expression via a p53-dependent signaling pathway. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Infrequent normal B lymphocytes express features of B-chronic lymphocytic leukemia.

1982 ◽  
Vol 155 (2) ◽  
pp. 623-628 ◽  
Author(s):  
F Caligaris-Cappio ◽  
M Gobbi ◽  
M Bofill ◽  
G Janossy
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Lymphocyte ◽  
Preliminary Evidence ◽  
Lymphocyte Subpopulation ◽  
Lymphocytic Leukemia ◽  
Phenotypic Characteristics ◽  
Human Tonsil ◽  
Hla Dr ◽  
Normal Human ◽  
Weak Expression

An infrequent (2-3%) B lymphocyte subpopulation was found in the normal human tonsil and lymph nodes that shows the phenotypic characteristics of B-chronic lymphocytic leukemia (B-CLL) (rosette formation with mouse erythrocytes, weak expression of membrane Ig, staining for HLA-DR, and OKT1 or Leu-1 detecting a T cell-associated p65 antigen). Preliminary evidence suggests that at least a subpopulation of these cells is found, in small proportions, within the germinal centers. These cells were not observed in the human bone marrow. B-CLL may involve this peripheral B lymphocyte subset.

scholarly journals Akt Inhibitors Induce Apoptosis in Chronic Lymphocytic Leukemia Cells.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1731-1731
Author(s):  
Mercè de Frias ◽  
Daniel Iglesias-Serret ◽  
Ana M Cosialls ◽  
Llorenç Coll-Mulet ◽  
Antonio F Santidrián ◽  
...  
Keyword(s):  
T Cells ◽  
Chronic Lymphocytic Leukemia ◽  
Conflicts Of Interest ◽  
Therapeutic Option ◽  
Mrna Level ◽  
Lymphocytic Leukemia ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Healthy Donors ◽  
Induced Apoptosis

Abstract Abstract 1731 Poster Board I-757 Phosphatidylinositol-3-kinase (PI3K)/Akt pathway has been described to be critical in the survival of chronic lymphocytic leukemia (CLL) cells. Here, we have analyzed the effect of two selective chemical inhibitors of Akt (Akti-1/2 and A-443654) in the survival of CLL cells. We studied by cytometric analysis the cytotoxic effects of Akt inhibitors on peripheral B and T lymphocytes from patients with CLL and from healthy donors. Both inhibitors induced apoptosis in CLL cells in a dose-dependent manner. Moreover, B cells from CLL samples were more sensitive to Akt inhibitors than T cells from CLL samples, and B or T cells from healthy donors. Survival factors for CLL cells, such as IL-4 and SDF-1a, were not able to block the apoptosis induced by both Akt inhibitors. We studied the changes induced by Akti-1/2 and A-443654 at mRNA level by performing reverse transcriptase multiplex ligation–dependent probe amplification (RT-MLPA). Akti-1/2 did not induce any change in the mRNA expression profile of genes involved in apoptosis, while A-443654 induced some changes, including an increase in NOXA and PUMA mRNA levels, suggesting the existence of additional targets for A-443654. We also studied the changes induced by both Akt inhibitors in some BCL-2 protein family members on CLL cells by Western blot. Both inhibitors induced an increase in PUMA and NOXA protein levels, and a decrease in MCL-1 protein level. Moreover, Akti-1/2 and A-443654 induced apoptosis irrespective of TP53 status. These results demonstrate that Akt inhibitors induce apoptosis of CLL cells and might be a new therapeutic option for the treatment of CLL. Disclosures No relevant conflicts of interest to declare.

scholarly journals Epidemiology Data of Patients with Chronic Lymphocytic Leukemia in West Central Mexico

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 5269-5269
Author(s):  
Virgina Campos-Cabrera ◽  
Gregorio Campos-Cabrera ◽  
Salvador Campos-Cabrera ◽  
Alicia Rivera-Trujillo ◽  
Sonia Hernandez-Rodriguez ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Conflicts Of Interest ◽  
Low Frequency ◽  
Epidemiological Data ◽  
Lymphocytic Leukemia ◽  
Female Ratio ◽  
Specific Protocol ◽  
Male Female Ratio ◽  
Epidemiology Data ◽  
International Data

Abstract Background: chronic lymphocytic leukemia (CLL) is a rare disease in Mexican mestizos (Br J Haematol 2015;169:909-911 and Int J Hematol 1999;69:253-255). No significant data on epidemiology is available. Methods: epidemiological data from samples referred to Laboratorios Fatima de Michoacan for flow cytometry immunophenotyping for neoplastic hematological disease. Results: 229 samples were received, 52 were diagnosed as CLL (22.7 %). Male 32 and female 20, ratio 1.6 a 1. Age from 33 to 89 years, average 66; 31 to 40 one, 41 to 50 two, 51 to 60 eleven, 61 to 70 twenty four, 71 to 80 ten, more than 81 four. Expression of CD 38 and ZAP-70 in three; only CD38 in 2, only ZAP-70 in three. Conclusions: similar results in male female ratio and age of presentation are noted as compared with international data. Low frequency of expression in CD38 and ZAP-70 may be due pre-analytic phase in the management of the sample. As a regional group we are trying to have epidemiology data in non-malignant and malignant hematological diseases to form specific protocol treatments. Figure 1. Figure 1. Disclosures No relevant conflicts of interest to declare.

scholarly journals Antigen presentation in an HLA-DR-restricted fashion by B-cell chronic lymphocytic leukemia cells

Blood ◽  
1988 ◽  
Vol 72 (1) ◽  
pp. 102-108 ◽  
Author(s):  
M Yasukawa ◽  
T Shiroguchi ◽  
A Inatsuki ◽  
Y Kobayashi
Keyword(s):  
T Cells ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Antigen Presentation ◽  
Interleukin 2 ◽  
Class Ii ◽  
Hla Class Ii ◽  
Lymphocytic Leukemia ◽  
Hla Dr ◽  
Cell Chronic Lymphocytic Leukemia

The ability of B-cell chronic lymphocytic leukemia (B-CLL) cells to present antigen to antigen-specific T cells was investigated. B-CLL cells present herpes simplex virus (HSV) antigen and purified protein derivative (PPD) to HSV- and PPD-specific, interleukin-2-dependent T- cell lines in an antigen-specific manner. Treatment of B-CLL cells with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) induced markedly increased levels of HLA-DR expression. TPA-treated B-CLL cells showed substantially more effective presentation, especially at low antigen concentrations, than did untreated B-CLL cells. By coculturing different allogeneic combinations of B-CLL cells and T cells and by adding anti-HLA-DR monoclonal antibody to cultures, it was found that antigen presentation by B-CLL cells was restricted by HLA-DR in the same way as for macrophages. We concluded from these experiments that B- CLL cells have a capacity to serve as antigen-presenting cells in an HLA class II-restricted fashion and that increasing the amount of HLA class II antigen and activation of B-CLL cells resulted in effective antigen presentation.

Differential Sensitivity of CD4+ and CD8+T Lymphocytes to the Killing Efficacy of Fas (Apo-1/CD95) Ligand+ Tumor Cells in B Chronic Lymphocytic Leukemia

Blood ◽  
1998 ◽  
Vol 91 (11) ◽  
pp. 4273-4281 ◽  
Author(s):  
Inge Tinhofer ◽  
Ingrid Marschitz ◽  
Marion Kos ◽  
Traudl Henn ◽  
Alexander Egle ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Cell Line ◽  
Tumor Cells ◽  
Fas Ligand ◽  
Lymphocytic Leukemia ◽  
Receptor Expression ◽  
Differential Sensitivity ◽  
Relative Reduction ◽  
Cd4 Cells ◽  
Fas Receptor

Abstract B-chronic lymphocytic leukemia (B-CLL) is characterized by cellular and humoral immune defects resulting in increased rates of infection and disturbed immune surveillance against cancer cells as well as by the expansion of slowly proliferating tumor cells. We found increased Fas receptor (FasR) expression in peripheral blood CD4+and CD8+ cells of B-CLL patients compared with the equivalent cells of healthy donors. Although increased Fas receptor expression was significant in both T-lymphocytic subsets, only CD4+ cells from B-CLL patients underwent apoptosis after treatment with the agonistic Fas antibody CH11. In CD4+cells of B-CLL patients, the Fas-sensitivity also correlated with a CD4+/CD8+ ratio below the lower threshold of healthy individuals (&lt;1.0). By contrast, FasR expression in the CD19+ fraction of B-CLL patients was downregulated compared with normal controls, and this was associated with an insensitivity to CH11-induced apoptosis. The B-CLL cell line EHEB as well as CD19+ cells from B-CLL patients constitutively expressed Fas ligand (FasL). The FasL was functionally active, as the B-CLL cell line as well as T-cell–depleted CD19+ B-CLL fractions were able to kill target T-acute lymphatic leukemia (T-ALL) cells in vitro. This effect was inhibited by the antagonistic FasR-antibody ZB4, the neutralizing anti-FasL monoclonal antibody (MoAb) NOK-2 or by transfection of the caspase inhibitor crmA. These data point to the fact that expression of FasL on CD19+B-CLL cells, together with enhanced susceptibility of CD4+ T cells toward FasL-bearing effector cells, are causally linked to the relative reduction of CD4+ cells occurring during B-CLL progression. These findings could explain the inversion of the ratio of CD4+/CD8+ cell numbers, which may be causally linked to the immune deficiency observed in these patients and to the expansion of the neoplastic clone in B-CLL.

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