Incidence, Sensitivity, and Specificity of Leukemia-Associated Phenotypes in Acute Myeloid Leukemia Using Specific Five-Color Multiparameter Flow Cytometry

Abstract

We assessed the usefulness of 5-color multiparameter flow cytometry to detect leukemia-associated phenotypes (LAPs) in the bone marrow of patients with newly diagnosed acute myeloid leukemia (AML) and determined its usefulness for detection of minimal residual disease (MRD). Overall, 94% of patients (51/54) with AML had LAPs at diagnosis. The frequency of leukemic bone marrow/median frequency of LAPs in normal or regenerating bone marrow samples using maximum log difference statistics revealed that CD2, CD56, CD11b, CD7, and CD19 expression on AML blasts represented the most sensitive and reliable markers for detection of MRD. Serial dilutional experiments showed that the sensitivity level of immunophenotyping was between 10-4 and 10-5 and that the approach was highly reproducible. Immunophenotypic analysis using a CD45 gating strategy, 5-color staining, and an extensive panel of monoclonal antibodies allowed the identification of LAPs in 94% of AML cases, and these immunophenotypes can be used for MRD monitoring with a sensitivity limit of 10-4 to 10-5.

Pre-transplant MRD predicts outcome following reduced-intensity and myeloablative allogeneic hemopoietic SCT in AML

“…[15][16][17] MRD monitoring by MFC relies on the detection of a patientspecific leukemia-associated immunophenotype at diagnosis by the co-expression of aberrant markers, which can be identified in 490% of the patients with AML if a comprehensive panel of MoAbs is employed. 18,19 Current techniques offer sensitivities between 1 Â 10 À 4 and 1 Â 10 À 5 16 and also offer the advantage of rapid analysis.…”

Clinical Significance of Co-expression of Aberrant Antigens in Acute Leukemia: A Retrospective Cohort Study in Makah Al Mukaramah, Saudi Arabia

“…Distinction between acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) is extremely important and flowcytometry (FCM) is very instrumental in this. Malignant blasts often have an abnormal phenotype that allows distinction from normal immature cells (Al-Mawali et al, 2008). One of these abnormalities is the occurrence of aberrant phenotypes (or anomalous expression) which is defined by the coexpression of markers usually not present on cells of that particular lineage (Ossenkoppele et al, 2011) e.g.…”

Enumeration and phenotypic assessment of human plasmacytoid and myeloid dendritic cells in whole blood

“…We have sourced the key antibodies and further improved the enumeration by introducing CD45 gating. CD45 gating on leucocytes has been shown by others to allow clearer and more sensitive gating strategies to be established (14)(15)(16). The panel has then been expanded to 6-colors, and evaluated against the 4-color panel.…”