Prognostic significance of CD38 and CD20 expression as assessed by quantitative flow cytometry in chronic lymphocytic leukaemia

Hsi, Eric D.; Kopecky, Kenneth J.; Appelbaum, Frederick R.; Boldt, David H.; Frey, Tom; Loftus, Margaret; Hussein, Mohamad A.

doi:10.1046/j.1365-2141.2003.04213.x

Cited by 44 publications

(25 citation statements)

References 36 publications

(75 reference statements)

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“…Quantitative Flow cytometry (QFCM) is being investigated as a diagnostic tool in the clinical laboratory and as a prognostic indicator in patients receiving antibodybased therapy (21)(22)(23)). In the current study, CD22 expression was quantitated by measuring the mean number of fluorescently-labeled antibodies bound per cell, or ABC, utilizing the QuantiBRITE assay and a custom conjugated anti-CD22 antibody with 1:1 PE to antibody ratio (F/P ratio 1:1).…”

Section: Discussionmentioning

confidence: 99%

“…The complete antibody panels, including antibody combinations used, were chosen based on the number of cells and previous histological diagnosis and immunophenotypic data. Erythrocytes were lysed by incubating with lysing solution (150 mM NH 4 Cl, 10 mM KHCO 3 , 0.1 mM EDTA) for 10 min at room temperature (maintained at [21][22][23] C) at a ratio of 1:9 (volume of sample: volume of lysing solution). Specimens were then washed with phosphate buffered saline (PBS) to remove cytophilic antibodies before determining cell number.…”

Section: Specimen Processingmentioning

confidence: 99%

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Variables affecting the quantitation of CD22 in neoplastic B cells

Jasper

Arun

Venzon

et al. 2010

Cytometry Part B Clinical

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Background: Quantitative flow cytometry (QFCM) is being applied in the clinical flow cytometry laboratory for diagnosis, prognosis, and assessment of patients receiving antibody-based therapy. ABC values and the effect of technical variables on CD22 quantitation in acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), follicular lymphoma (FCL), hairy cell leukemia (HCL) and normal B cells were studied.Methods: The QuantiBrite System V R was used to determine the level of CD22 expression (mean antibody bound per cell, ABC) by malignant and normal B cells. The intra-assay variability, number of cells required for precision, effect of delayed processing as well as shipment of peripheral blood specimens (delayed processing and exposure to noncontrolled environments), and the effect of paraformaldehyde fixation on assay results were studied.Results: The QuantiBRITE method of measuring CD22 ABC is precise (median CV 1.6%, 95% confidence interval, 1.2-2.3%) but a threshold of 250 malignant cells is required for reliable CD22 ABC values. Delayed processing and overnight shipment of specimens resulted in significantly different ABC values whereas fixation for up to 12 h had no significant effect. ABC measurements determined that CD22 expression is lower than normal in ALL, CLL, FCL, and MCL but higher than normal in HCL.Conclusions: CD22 expression was atypical in the hematolymphoid malignancies studied and may have diagnostic utility. Technical variables such as cell number analyzed and delayed processing or overnight shipment of specimens impact significantly on the measurement of antigen expression by QFCM in the clinical laboratory. Published

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Section: Discussionmentioning

confidence: 99%

Section: Specimen Processingmentioning

confidence: 99%

Variables affecting the quantitation of CD22 in neoplastic B cells

Jasper

Arun

Venzon

et al. 2010

Cytometry Part B Clinical

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“…A significant shorter time to first treatment and a shorter disease-specific survival was seen in patients with more than 250 ABC as compared to those with less than 250 ABC. Hsi et al (9) showed a significant shorter OS for patients with CD38 ABC >100, when compared to a group with <100 CD38 ABC. However, the complex patterns of CD38 expression by B-CLL cells in some patients prompted Ghia et al (10) to propose an approach in which they divided patients into three groups according to CD38 expression by the leukemic cells: those who were uniformly CD38 negative, those who were CD38 positive, and those with a bimodal profile (i.e., 2 distinct populations, CD38 negative and positive).…”

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confidence: 99%

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

Boonstra¹,

Lom²,

Langerak³

et al. 2006

Cytometry Part B Clinical

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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. q

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“…Tam et al (6) showed that CLL patients with trisomy 12 showed strong expression of CD20 and had a high rate of response to rituximab-based therapy. However, Hsi et al (7) reported that CD20 expression was not significantly correlated with clinical factors, and CD20 expression was also not associated with overall survival (OS). The prognostic significance of CD20 expression in B-cell precursor acute lymphoblastic leukemia has been extensively studied in children and adults.…”

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confidence: 99%

High levels of CD20 expression predict good prognosis in chronic lymphocytic leukemia

et al. 2013

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Heterogeneity of CD20 expression exists in chronic lymphocytic leukemia (CLL), therefore, we explored the prognostic significance of CD20 expression in Chinese patients with CLL. Multiparameter flow cytometry was used to detect the expression of CD20 in CD5 + CD19 + cells. In 172 CLL patients, the median expression percent of CD20 was 97.82% (range, 0-100), and the median mean fluorescence intensity (MFI) of CD20 in CLL cells was 731.45 (range, 0.00-9071.90). The percentage of CD20 + cells in the patient group with mutated variable region of immunoglobulin genes (IGHV) was higher than in the non-mutant IGHV group (mean, 92.1% vs 80.4%, P < 0.001). There were no differences in the MFI of CD20 + cells in all prognostic factor groups. Representation of the data using a receiver operating characteristic plot reflected separation between the two IGHV groups, with an area under the curve of 0.661 (95% confidence interval, 0.569-0.753). At the cut-off value of 60.3% for percentage of CD20, the sensitivity and specificity were 90.00% and 38.46%, respectively. Patients whose percentage of CD20 antigen was above 60.3% had longer treatment-free survival (hazard ratio, 0.452; 95% confidence interval, 0.232-0.884, P = 0.020). Percentage and MFI of CD20 were the variables not associated with treatment-free survival by multivariate Cox regression analysis (P < 0.05). High level of CD20 expression in de novo CLL appears to be associated with a good prognosis. (Cancer Sci 2013; 104: 996-1001) C hronic lymphocytic leukemia (CLL), the most frequently occurring leukemia in adults in North America and Europe, is a heterogeneous disease with variable clinical presentation and evolution.(1) In recent years, the significant increase in understanding the pathogenesis of CLL has been translated into a biologically oriented assessment of prognosis. In particular, there has been major progress in the identification of both molecular and cellular markers that may predict disease progression.(2,3) Two major molecular subtypes can be distinguished, characterized by a high or low number of somatic hypermutations in the variable region of immunoglobulin genes (IGHV) that features in CLL. However, IGHV gene sequencing remains a demanding technique, so many studies have focused on the identification of alternative markers with similar prognostic significance, and whose expression is easier to investigate, such as CD38 and zeta-associated protein-70 (ZAP-70). (4,5) A great deal of renewed interest in CD20 membrane protein has emerged since 1997, when the chimeric anti-CD20 mAb rituximab was approved for use in relapsed or refractory low-grade or follicular B-cell non-Hodgkin's lymphoma. Rituximab is commonly incorporated into CD20-positive B-cell lymphoma therapy, including CLL, to improve response and prognosis. The CD20 expression level might correlate with treatment outcome in CLL patients. Tam et al. (6) showed that CLL patients with trisomy 12 showed strong expression of CD20 and had a high rate of response to rituximab-based therapy. H...

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Prognostic significance of CD38 and CD20 expression as assessed by quantitative flow cytometry in chronic lymphocytic leukaemia

Cited by 44 publications

References 36 publications

Variables affecting the quantitation of CD22 in neoplastic B cells

Variables affecting the quantitation of CD22 in neoplastic B cells

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

High levels of CD20 expression predict good prognosis in chronic lymphocytic leukemia

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