Flow Cytometric Detection and Phenotype of Circulating CD34+ Cells in Patients Affected by Chronic Lymphocytic Leukemia.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4329-4329
Author(s):  
Fabio Stagno ◽  
Nunziata Laura Parrinello ◽  
Giovannella Fargione ◽  
Anna Triolo ◽  
Antonella Privitera ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Peripheral Blood ◽  
Flow Cytometric Analysis ◽  
Absolute Number ◽  
Lymphocytic Leukemia ◽  
Unexpected Finding ◽  
Advanced Stage ◽  
Flow Cytometric ◽  
Cd34 Cells ◽  
The Absolute

Abstract Flow cytometric determination of peripheral blood CD34+ cells provides reliable measurements of circulating hemopoietic progenitors. Since the detection of the absolute number of circulating CD34+ cells has been found of clinical utility in the setting of chronic myeloproliferative disorders, we investigated whether peripheral CD34+ cells could play any role in the clinical work-up of B-cell chronic lymphocytic leukemia (B-CLL). In this view, we determined by flow cytometry the absolute number of circulating CD34+ cells in the peripheral blood of 28 patients (16 males and 12 females, median age 67 years) affected by typical B-CLL (Matutes score 5,4,3) and in different Rai stages of the disease (19 early stage: Rai 0, I, II; 9 advanced stage: Rai III, IV). Conventional and multiparameter flow cytometric analysis was performed utilizing a FACSCalibur cytometer (Becton Dickinson). Our data showed a significant increase in the number of circulating CD34+ cells in the peripheral blood of patients with B-CLL (median CD34+ cells:7.8mL) as compared to controls (median CD34+ cells 0.1mL) (p=0.008). No statistical difference between B-CLL patients in early versus advanced stage (p=0.5) and between untreated versus treated (p=0.7) was found, as well as there was no correlation with some of the clinical characteristics of B-CLL (WBC-count, LDH levels, Beta-2M). In 10 out of 28 B-CLL affected patients, circulating CD34+ cells were correlated with ZAP-70 and CD38 antigen but no correlation was found. In addition, we detected in the peripheral blood of 22 out of 28 patients small numbers of circulating CD34+ cells displaying the CD19+/CD5+ phenotype (median CD34+/CD19+/CD5+ cells:5.7mL) whereas these cells were absent in normal controls. This unexpected finding, whose significance remains to be clarified and still restricted to a small number of cases, could be directly correlated to the underlying lymphproliferative disease and might represent a pool of leukemic stem cells. However, further studies are warranted.

scholarly journals Recombinant alpha 2-interferon in the treatment of B chronic lymphocytic leukemia in early stages

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1295-1298 ◽  
Author(s):  
C Rozman ◽  
E Montserrat ◽  
N Vinolas ◽  
A Urbano-Ispizua ◽  
JM Ribera ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Complete Response ◽  
Absolute Number ◽  
Peripheral Blood Lymphocytes ◽  
Response To Therapy ◽  
Lymphocytic Leukemia ◽  
Hematologic Toxicity ◽  
Low Doses ◽  
The Absolute ◽  
Cell Chronic Lymphocytic Leukemia

Abstract Ten previously untreated patients with early B cell chronic lymphocytic leukemia (B-CLL) (seven in Rai's stage 0, three in stage I) were given recombinant alpha 2-interferon (alpha 2IF) (2 X 10(6) U/m2 intramuscularly three times a week for a minimum of 14 weeks) to assess its effectiveness. All patients were evaluable for response to therapy and toxicity. No complete response was achieved. In all cases a definite, although transient reduction in the absolute number of peripheral blood lymphocytes was observed. In eight patients an increase in the absolute number of granulocytes was detected. None of the patients experienced severe hematologic toxicity. Fatigue, malaise, and fever were the more common side effects, but all patients were able to finish their treatment as planned. The results of this pilot study suggest that low doses of recombinant alpha 2-IF have some activity in early and previously untreated B-CLL and that further studies of IF effectiveness in B-CLL seem warranted.

P0934THE IMPACT OF HAEMODIALYSIS INITIATION ON CKD-ASSOCIATED MYELOID CELL DYSREGULATION

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Sarah Cormican ◽  
Serika Diane Naicker ◽  
Denise Connolly ◽  
Gemma Prendergast ◽  
M Conall Dennedy ◽  
...  
Keyword(s):  
Flow Cytometric Analysis ◽  
Myeloid Cell ◽  
Absolute Number ◽  
Flow Cytometer ◽  
Dialysis Initiation ◽  
Monocyte Subset ◽  
Cell Lineages ◽  
Flow Cytometric ◽  
The Absolute ◽  
Population Numbers

Abstract Background and Aims Abnormalities of the myeloid immune cell lineages are well described in people with both Chronic Kidney Disease (CKD) and End Stage Renal Disease (ESRD). Increased numbers and proportions of the intermediate (CD14++CD16+) and nonclassical (CD14+CD16+) monocyte subsets and increased neutrophil numbers have been reported in both settings. These changes are one aspect of the chronic inflammatory state which exists in people with CKD and ESRD and which contributes to complications including cardiovascular disease and protein-energy malnutrition. The effects of transition from advanced CKD to dialysis on myeloid cell populations has not been well studied. We investigated these changes by measuring major leucocyte population numbers and monocyte subset numbers and proportions in adults with Stage 5 CKD prior to and after haemodialysis initiation. Method Adults with stage 5 CKD and a non-emergent indication for dialysis initiation (in-centre haemodialysis) were recruited after informed consent. Blood samples were collected within the week prior to dialysis initiation and follow-up samples were collected prior to a dialysis session at one week and one month after haemodialysis initiation. Major leucocyte population numbers were determined by two-colour flow cytometric analysis of whole blood on an Accuri™ Flow Cytometer. Peripheral blood mononuclear cells were isolated and stained for multi-colour flow cytometric analysis on a Canto II™ flow cytometer. The absolute number of each monocyte subset/ml blood was calculated based on the above values. Results Ten individuals (six male, four female) with a mean age of 78 ± 6.4years were enrolled and completed all follow-ups. The mean eGFR was 9 ± 1.9ml/min at the time of haemodialysis initiation. Total monocyte numbers had not changed after one month of haemodialysis (Figure 1A) (3.6x105 ± 1.6x105cells/ml) compared to initiation (5.2x105 ± 3.2x105, p=0.11). However, the proportion of nonclassical monocytes was markedly increased after one week of haemodialysis (16.8% (IQR 12.8-21.0%) compared to 11.2% (IQR 9.3-12.3%) at initiation, p=0.007) (Figure 1B). At one month the proportion of nonclassical monocytes was maintained (17.1% (IQR 14.5-20.5%)) but had not increased further compared to the one week timepoint (p=0.97). This proportionate change was not reflected in the absolute nonclassical count. There were no significant changes in the proportion of classical or intermediate monocytes. Neutrophil numbers were reduced at one month (3.6x106 ± 1.3x106cells/ml) compared to initiation (4.8X106 ± 1.6x106cells/ml, p=0.04) (Figure 1A). Total lymphocyte numbers did not change significantly after dialysis initiation. Conclusion Haemodialysis initiation is associated with an increase in the proportion of nonclassical monocytes without significant increases in the absolute number of any monocyte subset. A reduction in total neutrophil number also occurs one month after dialysis initiation. It has been previously been shown that progression of CKD results in increasing abnormalities of the myeloid cell lineages. Here we have demonstrated that transition from advanced CKD to haemodialysis results in further modulation of myeloid cell numbers and myeloid cell subset proportions.

Receptor Tyrosine Kinase-Like Orphan Receptor 1 (ROR1) - a Putative Diagnostic Marker for Chronic Lymphocytic Leukemia (CLL).

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1587-1587
Author(s):  
Sabrina Uhrmacher ◽  
Magdalena Hertweck ◽  
Julian Paesler ◽  
Felix Erdfelder ◽  
Alexandra Filipovich ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Chronic Lymphocytic Leukemia ◽  
Tyrosine Kinase ◽  
Healthy Volunteers ◽  
Receptor Tyrosine Kinase ◽  
Peripheral Blood ◽  
Lymphocytic Leukemia ◽  
Orphan Receptor ◽  
Flow Cytometric

Abstract Abstract 1587 Poster Board I-613 In chronic lymphocytic leukemia (CLL) WNT signaling is constitutively active and several members of this signaling pathway are uniformely upregulated in these cells. Apart from classical WNT receptors like FZD and LRP6, receptor tyrosine kinase-like orphan receptor 1 (ROR1) has been shown to function as a receptor for WNT proteins, too. Furthermore, it could recently be demonstrated that ROR1 is frequently expressed on the surface of CLL cells and might therefore serve as a therapeutic target in this disease. However, so far only little is known about the expression status of this protein in different patients. Moreover, a diagnostic antibody for flow cytometric investigations is lacking. Thus, the aim of our study was to i) establish a directly labelled anti-ROR1 antibody for flow cytometry, ii) to confirm previous results on ROR1 expression in CLL, iii) to investigate ROR1 expression in different cell compartments and iv) correlate our findings to known markers of risk and disease progression. Peripheral blood of CLL patients as well as healthy volunteers was subjected to flow cytometric analysis. Besides standard determination of leukocyte subpopulations ZAP70 and CD38 status was assessed according to current diagnostic recommendations. In addition, ROR1 surface expression was first detected by flow cytometry using a specific primary antibody directed against ROR1 and a fluorescent labelled secondary antibody. Using this experimental setting we found that ROR1 is expressed on 63.4% of all neoplastic CLL cells and also on 30.5% of T cells in the peripheral CLL blood. In contrast, no ROR1 expression could be detected on NK cells, B cells, CD8+- or CD4+-T cells of healthy individuals. To improve the analytical technique the ROR1 antibody was directly conjugated with Phycoerythrin (PE) and the experiments were repeated. With the conjugated antibody we detected ROR1 expression on 97.1% of neoplastic CLL cells and virtually on no T lymphocytes. ROR1 expression levels correlated neither with the expression of ZAP70 nor with CD38. Again, we could not detect ROR1 expression on peripheral blood cells of our healthy volunteers. Taken together, ROR1 expression appears to be highly restricted to CLL cells. If in addition to CD5 and CD19 ROR1 detection is included into diagnostic flow cytometric panels the specificity and sensitivity of immunophenotypic CLL diagnostics may be greatly enhanced. Disclosures Hallek: Roche: Consultancy, Honoraria, Research Funding.

CD49d Expression Identifies a Chronic Lymphocytic Leukemia (CLL) Subset with High Levels of Circulating CD34 +Cells Co-Expressing Endothelial Cell Markers.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2329-2329
Author(s):  
Francesca Maria Rossi ◽  
Davide Rossi ◽  
Antonella Zucchetto ◽  
Riccardo Bomben ◽  
Michele Dal Bo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Chronic Lymphocytic Leukemia ◽  
Lymphocytic Leukemia ◽  
Bone Marrow Biopsies ◽  
Endothelial Markers ◽  
Cd38 Expression ◽  
Cell Counts ◽  
Cd34 Cells ◽  
Stromal Component ◽  
The Absolute

Abstract Abstract 2329 Poster Board II-306 Introduction: In chronic lymphocytic leukemia (CLL), CD49d, often in association with CD38, has been shown to mark a disease subset with poor prognosis. Functionally, both molecules act as counter-receptors for surface structures (i.e. VCAM-1/CD106 and CD31) usually expressed by the endothelial/stromal component of tumor micro-environment. We have recently identified a micro-environmental circuitry which involves CD38 triggering, and eventually determines an enrichment of the VCAM-1/CD106-expressing endothelial component detected in the context of CLL infiltrates found in bone marrow biopsies. Data was also provided that CD49d/VCAM-1 interactions are active in delivering pro-survival signals to CD49d-expressing CLL cells (Zucchetto et al, Cancer Res, 69, 4001, 2009). In this study, we investigated the amount of circulating progenitors with endothelial phenotype in CLL samples with different CD49d and CD38 expression levels. Methods: Peripheral blood (PB) samples from 91 CLL cases purposely selected with WBC>25,000/μl (B cells absolute lymphocyte count >10,000/μl) were evaluated by multiparametric flow cytometry for the absolute count of circulating CD34+ cells (ISHAGE protocol in single platform). Whenever possible (i.e. if a cluster of at least 100 CD34+ cells was detectable), a further characterization was performed (4-6 colours flow cytometry) for circulating endothelial cells (CEC), identified as a CD34+CD45low cell population co-expressing one of the following endothelial markers: CD309/VEGFR-2, CD144/VE-cadherin, CD106/VCAM-1 and CD146/Muc-18. CD49d and CD38 expression by CLL cells was considered positive if exceeding the standard cut-off value of 30% of positive cells. Results: PB absolute CD34+ cell counts were 7.5±7.5/μl in CD49d+ CLL (32 cases), vs. 3.3±2.7/μl in CD49d− CLL (59 cases; p=2.6×10−4), or 9.4±8.7/μl in CD49d+ CLL (30 cases) vs. 4.6±2.9/μl in CD49d− CLL (18 cases; p=0.004) when only cases phenotyped for CEC were considered. Furthermore, when samples were stratified also for CD38 expression, values of circulating CD34+ cells increased to 10.6±10.1/μl in CD38+CD49d+ CLL (11 cases) vs. 3.1±2.4/μl in CD38−CD49d− (51cases; p=1×10−5). Regarding the absolute quantification of CEC, a CD49d+ phenotype again marked the CLL subset with the highest CEC count, as identified by the expression of either the CD309/VEGFR-2 (CEC counts 1.7±2.3/μl in CD49d+ CLL vs. 0.5±0.5/μl CD49d− CLL; p=0.009) or the CD144/VE-cadherin (CEC counts 0.8±1/μl in CD49d+ CLL vs. 0.3±0.5/μl in CD49d− CLL; p=0.057) endothelial markers on CD34+CD45low cells. Notably, CEC from CD49d+ CLL expressed CD106/VCAM-1 in virtually all cells (1.6±2.4/μl), while the other marker of endothelial activation CD146/Muc-18 was detected in a fraction of CEC only (0.4±0.9/μl). Conclusions: CD49d and CD38 expression by CLL cells identify a disease subset with significantly higher number of both circulating CD34+ cells and CEC. This phenomenon could be explained considering several aspects: i) the sharing of common phenotypic markers between CLL cells and CD34+ progenitors, including CD38 and CD49d, which could be responsible for a displacement of CD34+ progenitors in the context of micro-environmental niches; ii) the known capacity of CLL cells, especially with a unmutated IGHV gene status and/or a CD38+CD49d+ phenotype to produce pro-angiogenic factors including Ang-2; iii) the rare PB cells expressing CD34 and CEC markers may represent CLL cell precursors with tumor-initiating cell features. Studies are currently ongoing to dissect among these hypotheses Disclosures: No relevant conflicts of interest to declare.

scholarly journals Recombinant alpha 2-interferon in the treatment of B chronic lymphocytic leukemia in early stages

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1295-1298
Author(s):  
C Rozman ◽  
E Montserrat ◽  
N Vinolas ◽  
A Urbano-Ispizua ◽  
JM Ribera ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Complete Response ◽  
Absolute Number ◽  
Peripheral Blood Lymphocytes ◽  
Response To Therapy ◽  
Lymphocytic Leukemia ◽  
Hematologic Toxicity ◽  
Low Doses ◽  
The Absolute ◽  
Cell Chronic Lymphocytic Leukemia

Ten previously untreated patients with early B cell chronic lymphocytic leukemia (B-CLL) (seven in Rai's stage 0, three in stage I) were given recombinant alpha 2-interferon (alpha 2IF) (2 X 10(6) U/m2 intramuscularly three times a week for a minimum of 14 weeks) to assess its effectiveness. All patients were evaluable for response to therapy and toxicity. No complete response was achieved. In all cases a definite, although transient reduction in the absolute number of peripheral blood lymphocytes was observed. In eight patients an increase in the absolute number of granulocytes was detected. None of the patients experienced severe hematologic toxicity. Fatigue, malaise, and fever were the more common side effects, but all patients were able to finish their treatment as planned. The results of this pilot study suggest that low doses of recombinant alpha 2-IF have some activity in early and previously untreated B-CLL and that further studies of IF effectiveness in B-CLL seem warranted.

Clinical Predictors of Abnormal Peripheral Blood Lymphocytoses Diagnosed by Flow Cytometry: An Algorithmic Approach That Can Be Applied in Routine Clinical Practice.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3932-3932
Author(s):  
Jared M. Andrews ◽  
Mitchel T. Holm ◽  
Jerome B. Myers
Keyword(s):  
Flow Cytometry ◽  
Peripheral Blood ◽  
Lymphocyte Count ◽  
Flow Cytometric Analysis ◽  
Clinical History ◽  
Lymphocytic Leukemia ◽  
Western World ◽  
Abnormal Phenotype ◽  
Flow Cytometric ◽  
The Mean

Abstract Background Elevated peripheral blood lymphocyte counts in adults can occur in benign reactive conditions as well as malignant disease processes. Chronic lymphocytic leukemia (CLL) is the most common adult hematologic malignancy of the western world affecting the middle aged and elderly. Less commonly B, T, and Natural Killer (NK) cell leukemia / lymphomas may also present with lymphocytosis. Flow cytometry has greatly improved the ability to detect low levels of abnormal lymphocyte populations in peripheral blood. It is, however, a relatively expensive test and clinical guidelines for its appropriate usage are not well defined. Methods We conducted a retrospective review of peripheral blood lymphocytoses that were submitted for flow cytometric analysis at Madigan Army Medical Center, Tacoma, WA from 2002 – 2004. Under laboratory protocol, all patients ≥ 50 years of age with an absolute lymphocyte count (ALC) of > 4 X 109 Cells/L had a peripheral smear evaluated by both a hematology technician and pathologist. Specimens determined to warrant flow cytometric analysis based on review of clinical history, prior lab values, degree of lymphocytosis, and morphology were either recommended for flow cytometry in a comment; or sent directly for analysis with the clinician’s approval. We reviewed complete blood counts (CBCs), previous flow cytometry results, as well as bone marrow and electronic clinical history. All patients with previous diagnoses of lymphoproliferative disorders (LPDs) or ALC < 4 X 109 Cells/L were excluded. Results Approximately 7,300 CBC specimens/month (3,400 from patients ≥ 50 years of age) were performed. Of these, an average of 44 specimens/month had a lymphocytosis of ≥ 4 X 109 Cells/L, from approximately 28 different patients. From this group 71 flow cytometric cases (an average of 2/month) were performed over the 2 year period. 42 cases (59%) had an abnormal phenotype. 27 had a phenotype consistent with CLL, and the other 15 were a mixture of LPDs involving B and T-lymphocytes as well as NK cells. Comparing normal phenotype to abnormal phenotype showed statistically significant differences between the mean age (n-60.4 ±7.5, abn-69.8±8.7), ALC (n-4.9±0.8, abn-9.2±8.1), and relative lymphocyte count (RLC) (n-43.9±7.5%, abn-59.3±8.8%). Conclusion Absolute lymphocyte counts ≥ 4 X 109 Cells/L in adults ≥ 50 years of age represent approximately 1% of the CBCs performed in our laboratory. Review of these cases by a pathologist is logistically feasible due to the low incidence. Our method of reviewing for morphology, clinical history, and past lymphocyte counts with comments to the ordering clinician yielded a high incidence of abnormal phenotype diagnoses when evaluated by flow cytometric analysis (59%). Age, ALC, and relative lymphocyte counts are variables that can be used to develop guidelines for determining the appropriateness of flow cytometric analysis. Patients < 52.4 years of age fall below two standards of deviation from the mean age of the abnormal phenotype group. The standard of deviation for mean ALC is very small (4.9±0.8), which indicates that counts > two standards of deviation above the mean, or 6.5 X 109 Cells/L, would correlate strongly with an abnormal phenotype. The same conclusion could be made with a RLC > 58.9%. In conclusion, patients ≥ 50 years of age with an ALC > 6.5 X 109 Cells/L or a RLC > 58.9% are likely to have a lymphoproliferative disorder and flow cytometric analysis is indicated.

Phase IIa Study of the Anti-CXCL12/SDF-1 Spiegelmer® Nox-A12 Alone and Combined with Bendamustine/Rituximab in Patients with Relapsed Chronic Lymphocytic Leukemia (CLL)

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4593-4593
Author(s):  
Marco Gobbi ◽  
Federico Caligaris-Cappio ◽  
Marco Montillo ◽  
Stephanie Vauléon ◽  
Stefan Zöllner ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Plasma Concentration ◽  
Chronic Lymphocytic Leukemia ◽  
Healthy Volunteers ◽  
Peripheral Blood ◽  
Combined Treatment ◽  
Single Agent ◽  
Lymphocytic Leukemia ◽  
Leukemic Cells ◽  
Cd34 Cells

Abstract Abstract 4593 Background NOX-A12 is a novel, potent, L-aptamer inhibitor of CXCL12/SDF-1, a chemokine which attracts and activates immune- and non-immune cells. The signaling of CXCL12 has been shown to play an important role in the pathophysiology of chronic lymphocytic leukemia (CLL), especially in the interaction of leukemic cells with their tissue microenvironment. The therapeutic concept of NOX-A12 is to inhibit such tumor-supporting pathways and thereby sensitizing the CLL cells towards chemotherapy. Methods The purpose of this phase IIa study is to evaluate the safety and efficacy of NOX-A12 in combination with background chemo-immunotherapy of bendamustine and rituximab (BR) in patients with relapsed CLL. The described study is being performed in compliance with ethical principles based on the Declaration of Helsinki and ICH-GCP guidelines. The study population was split into a pilot and expansion group. In the pilot group, 3 cohorts of 3 patients each received escalating doses of single agent NOX-A12 two weeks prior to the combined treatment of NOX-A12 and BR. Interim data from these patients are reported. Based on previous Phase I studies in healthy volunteers, pilot patients received a dose of 1, 2 or 4 mg/kg body weight (BW) single agent NOX-A12 on day -14, followed by a 2-weeks period of safety, PK and PD assessments prior to the combined treatment with NOX-A12 and BR. To date, the first cohort of the pilot group already progressed to the 2nd cycle of combined treatment. Evaluation criteria included adverse events according to CTCAE V4, flow cytometry of peripheral blood CD34+ cells and CLL cells, pharmacokinetics of NOX-A12, plasma concentration of CXCL12 and tumor response (NCI-WG 1996 criteria, updated 2008). Results To date 3 patients (age range: 58 – 65 years) have been enrolled in the pilot group of this study. They had received 1 or 2 prior therapies, but no bendamustine. Single i.v. doses of 1 mg/kg BW NOX-A12 had no clinically relevant effects on vital signs, 12-lead ECG parameters and laboratory parameters. One patient reported grade 1 pain in the lower limbs two days after treatment with NOX-A12. This event was not dose-limiting and resolved spontaneously on the same day. Flow cytometry of CD34+ cells and CLL cells (CD19+/CD5+high) showed a rapid mobilization of these cells into the peripheral blood on day 1. Interestingly, return to baseline was not complete at the last assessment on day 3 (for details see Figure 1). The NOX-A12 pharmacokinetics in these 3 patients (for concentration-time profile see Figure 2) is very comparable to healthy volunteers receiving i.v. NOX-A12, with a maximum plasma concentration of 1.52 ± 0.14 μM after 1 h (tmax) and a plasma elimination half-life of about 50 h. As seen in healthy volunteers the plasma concentration of CXCL12 increased upon NOX-A12 treatment and reached a maximum of 0.434 ± 0.076 μM at 24 to 72 h p.a. without ever approaching the plasma concentration of NOX-A12 (Figure 2). Conclusion Single i.v. doses of NOX-A12 at 1 mg/kg BW were safe and well tolerated; the maximum tolerated dose was not reached. NOX-A12 induced a long-lasting mobilization of CD34+ cells and leukemic cells in patients with relapsed CLL, consistent with a mechanism of action based on CXCL12 inhibition. Patient accrual and identification of an optimal chemosensitization regimen of NOX-A12 combined with BR is being continued. Disclosures: Vauléon: NOXXON Pharma AG: Employment. Zöllner:NOXXON Pharma AG: Employment. Dümmler:NOXXON Pharma AG: Employment. Kruschinski:NOXXON Pharma AG: Employment. Fliegert:NOXXON Pharma AG: Employment.

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