Related Experiment Video
Updated: Aug 11, 2026
Comprehensive Protocol to Sample and Process Bone Marrow for Measuring Measurable Residual Disease and Leukemic Stem Cells in Acute Myeloid Leukemia
Published on: March 5, 2018
Leukemia markers expression of peripheral blood vs bone marrow blasts using flow cytometry
Abbas Rezaei1, Minoo Adib, Fariborz Mokarian
1Depertment of Immunology, School of Medicine, University of Medical Sciences, Isfahan, Iran. rezaei@mui.ac.ir
Insights
Flow cytometry immunotyping of peripheral blood (PB) and bone marrow (BM) shows strong correlation in acute myeloid leukemia (AML) patients. This finding may reduce the need for invasive BM aspiration in leukemia diagnosis.
Area of Science:
- Hematology
- Immunology
- Oncology
Background:
- Flow cytometry is crucial in clinical hematology for leukemia characterization.
- Immunotyping aids in identifying leukemia subtypes and prognostic indicators.
- Morphological interpretation can be challenging, making immunotyping valuable.
Purpose of the Study:
- To compare immunophenotypic marker expression in peripheral blood (PB) and bone marrow (BM) samples of acute leukemia patients.
- To evaluate the utility of flow cytometry in diagnosing acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL).
- To determine if PB analysis can obviate the need for BM aspiration.
Main Methods:
- Evaluated 18 AML and 13 ALL patients with ≥30% blasts in PB.
- Collected PB and BM samples for flow cytometry analysis.
- Utilized a panel of monoclonal antibodies targeting various cell lineages and markers (e.g., CD markers, TdT, HLA-DR, CD45).
Main Results:
- Discrepancies in marker expression were observed in the ALL group.
- A strong correlation in marker expression between PB and BM samples was found in AML patients for most tested markers.
- Specifically, CD13, CD14, CD45, and HLA-DR showed positive correlation, while CD3, CD5, CD13, CD14, CD19, CD45, HLA-DR, and TdT showed strong correlation in AML.
Conclusions:
- Targeted gating and appropriate antibody panels are essential for consistent leukemia immunotyping in PB and BM.
- Preliminary findings suggest that PB immunophenotyping may minimize the necessity of bone marrow aspiration for leukemia diagnosis.
- Accurate immunotyping in PB can aid in treatment decisions and prognosis.
Background:
Flow cytometric techniques are widely used in clinical hematology. Characterization of leukemias by immunotyping is particularly helpful when the morphology is difficult to interpret. The major advantage of using immune markers by flow cytometry is the identification of particular leukemia subtype, not recognized by morphologic criteria, which may have prognostic significance. Current literature suggests when peripheral blood (PB) is consisted of 30% blasts or higher diagnosis of acute leukemia is most likely. However, bone marrow aspiration may also be performed as a confirmatory diagnosis. Immunotyping of PB and BM in leukemias not only determine the decision making for a specific therapeutic regimen, but also is a practical prognostic indicator.
Material/Methods:
We evaluated 18 patients with acute myeloid Leukemia (AML) and 13 patients with acute lymphoid leukemia (ALL). In all cases, the amount of blasts in PB was 30% or higher. Two ml PB and BM samples from each patient was collected. Following the preparation process, expression of markers was detected by using flow cytometry. The panel of monoclonal antibodies used in this study were consisted of CD3, CD7, CD5 (for T lymphocytes lineage); CD19, CD22, CD20, CD10 (for B lymphocytes lineage); CD13, CD14, CD33 (for myeloid subsets); and TDT, HLA-DR, CD45 (non lineage restricted). Expression levels of PB and BM markers were compared by using statistical analysis.
Results:
The results showed apparent discrepancies for some markers in ALL group. However, in AML patients most of the selected markers have shown considerable correlation between PB and BM samples. Only four markers (CD13, CD14, CD45, and HLA-DR) showed positive correlation. In contrast, most markers (CD3, CD5, CD13, CD14, CD19, CD45, HLA-DR, and TdT) showed strong correlation between PB and BM samples in AML group.
Conclusions:
The findings of this study suggests that targeted gating strategy for blast population as well as selection of a suitable panel of monoclonal antibodies may be essential for diagnosis of leukemia resulting in similar immunotyping pattern in PB and BM. Although our results are preliminary, this can minimize the necessity of BM aspiration for leukemia patients.
More Related Videos
09:01Flow Cytometry to Estimate Leukemia Stem Cells in Primary Acute Myeloid Leukemia and in Patient-derived-xenografts, at Diagnosis and Follow Up
Published on: March 26, 2018
08:34Flow Cytometry Analysis of Murine Bone Marrow Hematopoietic Stem and Progenitor Cells and Stromal Niche Cells
Published on: September 28, 2022