U.S.-Canadian consensus recommendations on the immunophenotypic analysis of hematologic neoplasia by flow cytometry: Data analysis and interpretation

Borowitz, Michael J.; Bray, Robert A.; Gascoyne, Randy D.; Melnick, Steven J.; Parker, Jōhn W.; Picker, Louis J.; Stetler‐Stevenson, Maryalice

doi:10.1002/(sici)1097-0320(19971015)30:5<236::aid-cyto4>3.0.co;2-f

Cited by 122 publications

(68 citation statements)

References 78 publications

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“…Antibody positivity was determined by comparison of the shape of either histograms or dot plots to those of control nonreactive antibodies. 19 Initially, each antibody was categorized simply as positive or negative based on whether the histogram showed a significant shift (usually a minimum of 80-100 channels on a 4 decade log scale) relative to nonreactive antibodies, or if a distinct subset of the gated blasts, generally amounting to at least 10% of cells, showed expression that was clearly above the threshold determined by consideration of the reactivity of the control antibody. In subsequent analyses, these two patterns were distinguished, and cases in which only a subset of cells were seen to be positive was referred to as having 'partial' positivity.…”

Section: Immunophenotypingmentioning

confidence: 99%

Surface antigen phenotype can predict TEL-AML1 rearrangement in childhood B-precursor ALL: a Pediatric Oncology Group study

et al. 1998

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The t(12;21)(p13;q22) is the most common translocation in childhood B-precursor ALL. It results in a TEL-AML1 rearrangement and is associated with a good prognosis. Because many chromosomal alterations in leukemia are associated with distinct cell surface phenotypes, we investigated whether there was an association seen between surface marker expression and the TEL-AML1 rearrangement. Of 166 unselected cases of B-precursor ALL studied by Southern hybridization, 45 cases (27%) showed TEL rearrangement. Blasts of patients with TEL rearrangement were much more likely to be CD9-negative, CD45-positive, CD13 positive, and CD20 negative, but the predictive value of any of these markers for the rearrangement was very low. However, 93% of patients with the TEL rearrangement had blasts that were either negative or only partly positive for CD9; this phenotype was only seen in 27% of patients without the rearrangement. Only information about CD20 expression added to the predictive value of CD9 alone. The predictive value of the phenotype CD9 (negative or partly positive) and CD20 (negative or partly positive), for the TEL rearrangement was prospectively tested on an additional 223 cases, and found to be 88% sensitive and 71% specific for the rearrangement, with a positive predictive value of 47%. Hyperdiploidy, previously shown to correlate negatively with the rearrangement, was a slightly more sensitive indicator (94%) but had a much lower predictive value (28%). Three of eight cases found to be rearranged by Southern hybridization but lacking the characteristic phenotype failed to show evidence of the TEL-AML1 rearrangement by polymerase chain reaction, suggesting that at least some of the discordant cases may involve partner genes other than AML1 in the TEL rearrangement. We conclude that immunophenotyping is highly predictive of the TEL rearrangement. For every 100 patients with B-precursor ALL, we estimate that prescreening by phenotyping would eliminate the need for molecular testing on 57 patients and only two or three of an expected 24 patients with the TEL rearrangement would not be detected. Keywords: childhood; acute lymphoblastic leukemia; TEL; AML1; flow cytometry IntroductionThe TEL-AML1 fusion, created by a cryptic t(12;21), is the most common translocation in childhood ALL, occurring in about 25% of cases. [1][2][3] Patients with this translocation have a favorable prognosis.4-6 Blast cell hyperdiploidy, another important favorable prognostic factor, is negatively correlated with the presence of this translocation, 2,5,6 so that these two favorable genetic characteristics identify separate subsets of children. Although the TEL-AML1 translocation is restricted to patients with B-precursor ALL, and specifically to common ALL, 4,6 relatively little additional information is available about the blast cell characteristics of these cases. 6 Cytogenetic abnormalities in both AML and ALL may be associated with distinct surface antigen phenotypic properties 7-14 These distinctive phenotypic properties are sometimes ...

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Section: Immunophenotypingmentioning

confidence: 99%

Surface antigen phenotype can predict TEL-AML1 rearrangement in childhood B-precursor ALL: a Pediatric Oncology Group study

et al. 1998

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“…We provide recommendations for the use of controls to identify background fluorescence in clinical flow cytometry such as unstained cells, cells that do not express the antigen of interest in an antibody-labeled sample, as well as fluorescence-(or full)-minus-one (FMO), isotype and isoclonic controls. In addition to previously published guidelines (6)(7)(8)(9), this document is intended to contribute to formulation of consensus regarding the utility of flow cytometry for analysis of clinical samples.…”

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Considerations for the control of background fluorescence in clinical flow cytometry

Hulspas¹,

O’Gorman

Wood

et al. 2009

Cytometry Part B Clinical

209

190

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“…[1][2][3][4][5] In a clinical setting, the main purpose of flow cytometric analysis is to identify abnormal cells by defining their immunophenotypic characteristics. To accomplish this task, multiparameter analysis represents a reliable approach that integrates the information provided by forward scatter (FSC) and side scatter (SSC) with multiple fluorescence parameters.…”

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confidence: 99%

“…9 This strategy is also successfully applied for the analysis of many lymphoid malignancies detected in blood, bone marrow, and lymph nodes. 1,9 Specific immunophenotypic profiles have been established for several lymphoid neoplasms. [1][2][3][4] The abnormal profiles are compared to the pattern of cell marker expression normally seen in the organ/tissue being examined, and a diagnostic interpretation is made.…”

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confidence: 99%

“…1,9 Specific immunophenotypic profiles have been established for several lymphoid neoplasms. [1][2][3][4] The abnormal profiles are compared to the pattern of cell marker expression normally seen in the organ/tissue being examined, and a diagnostic interpretation is made. While the normal reference ranges for differentiation antigens expressed by lymphoid cells from peripheral blood, bone marrow, and lymph nodes are well established, [11][12][13][14][15] there is little information concerning the immunophenotypic profile of lymphocytes found in normal or reactive spleen.…”

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Flow cytometric analysis of normal and reactive spleen

Colovai

Giatzikis

et al. 2004

Modern Pathology

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Spleen is surgically removed for both non-neoplastic and neoplastic pathologies. A significant proportion of splenectomy specimens require distinguishing between reactive and neoplastic conditions (eg lymphoma). To establish a 'normal' reference range for the spleen lymphocyte subsets, fresh samples of benign, reactive spleens obtained from adult patients (N ¼ 12) and samples of normal spleen obtained from cadaveric transplant donors (N ¼ 14) were analyzed using three-and four-color flow cytometry. Study of pan-B, -T, and -NK marker expression revealed that the frequency of T cells is higher and that of B cells is lower in reactive (nonneoplastic) compared to normal (cadaveric) spleen. Furthermore, our study established a frame of reference for cell markers commonly used for immunophenotyping of lymphoma, and identified discrete lymphocyte subsets, such as early plasma cells and T cells carrying the phenotype of the NK/T subset. These results will facilitate an accurate interpretation of the flow cytometric analysis of human spleen lymphocytes.

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U.S.-Canadian consensus recommendations on the immunophenotypic analysis of hematologic neoplasia by flow cytometry: Data analysis and interpretation

Cited by 122 publications

References 78 publications

Surface antigen phenotype can predict TEL-AML1 rearrangement in childhood B-precursor ALL: a Pediatric Oncology Group study

Surface antigen phenotype can predict TEL-AML1 rearrangement in childhood B-precursor ALL: a Pediatric Oncology Group study

Considerations for the control of background fluorescence in clinical flow cytometry

Flow cytometric analysis of normal and reactive spleen

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