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Normal lymphocytes from leukemic samples as an internal quality control for fluorescence intensity in immunophenotyping of acute leukemias
Author(s) -
Ratei Richard,
Karawajew Leonid,
Lacombe Francis,
Jagoda Krystyna,
Del Poeta Giovanni,
Kraan Jaco,
De Santiago Maria,
Kappelmayer János,
Björklund Elisabeth,
Ludwig W.D.,
Gratama Jan,
Orfao Alberto
Publication year - 2005
Publication title -
cytometry part b: clinical cytometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 61
eISSN - 1552-4957
pISSN - 1552-4949
DOI - 10.1002/cyto.b.20075
Subject(s) - immunophenotyping , cd3 , medicine , leukemia , acute lymphocytic leukemia , flow cytometry , minimal residual disease , pathology , antigen , lymphoblastic leukemia , immunology , cd8
Background Multiparametric flow cytometry has become an indispensable but complex tool for the diagnosis of acute leukemias. Interpretation of immunophenotypic data within a six‐parameter analytical space relies on the standardization and validation of the instrument, the reagents, and the procedure. To address whether or not residual normal lymphocytes, usually present within leukemic samples, can serve as internal quality control for fluorescence intensity, 116 leukemic and 35 normal samples were analyzed. Methods Eight laboratories participated in the study and recruited a total of 151 individuals including 29 patients with B‐cell precursor acute lymphoblastic leukemia (BCP‐ALL), 77 with acute myeloid leukemia (AML), 10 with T‐cell precursor acute lymphoblastic leukemia (T‐ALL), and 35 normal bone marrow donors. Lymphocytes were gated according to the CD45 hi /SSC lo gating strategy, after which median fluorescence intensities (MFI) as well as percentages of positive cells (%positive) for CD19, CD22, CD7, and CD3 were recorded. Nonparametric statistics were used to compare variation within and between laboratories. Results Normal lymphocytes within leukemic samples do not show substantial differences compared to lymphocytes from normal controls with respect to expression of CD19, CD22, CD7, and CD3. In particular, longitudinal control charts of MFI values for CD3 antigen provide useful information on analytical and instrument performance. Conclusion Residual normal lymphocytes can serve as internal quality control for studies addressing fluorescence intensity in the setting of immunophenotyping of acute leukemias. © 2005 Wiley‐Liss, Inc.