Related Experiment Video
Updated: Aug 10, 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
Flow cytometric analysis of cell-surface and intracellular antigens in leukemia diagnosis
Insights
New flow cytometry technology enables sensitive detection of intracellular antigens for identifying early blood cell differentiation stages. This method is crucial for diagnosing acute myeloblastic leukemia (AML) by analyzing myeloperoxidase (MPO) and other markers.
Area of Science:
- Hematology
- Immunology
- Flow Cytometry
- Leukemia Diagnostics
Background:
- Accurate identification of early hematopoietic differentiation is essential for diagnosing hematologic malignancies.
- Traditional methods may lack sensitivity or specificity in distinguishing early myeloid and lymphoid lineages.
- Intracellular antigens offer potential for more precise lineage and differentiation assessment.
Purpose of the Study:
- To evaluate a new, highly sensitive flow cytometric technology for analyzing intracellular antigens in hematopoietic cells.
- To demonstrate the utility of this technology in identifying early differentiation stages of myeloid and lymphoid cells.
- To assess the application of intracellular myeloperoxidase (MPO) and lactoferrin (LF) for acute myeloblastic leukemia (AML) diagnosis.
Main Methods:
- Utilized highly sensitive flow cytometry for intracellular antigen detection and quantitative analysis.
- Employed antibodies against intracellular myeloperoxidase (pro-)MPO, CD22, and CD3 for lineage identification.
- Combined MPO staining with lactoferrin (LF) staining to differentiate maturation compartments in bone marrow.
Main Results:
- Successfully identified earliest myeloid and lymphoid differentiation stages using intracellular MPO, CD22, and CD3.
- Demonstrated the high relevance of intracellular MPO for diagnosing undifferentiated AML, outperforming other myeloid markers.
- Showcased that combining MPO and LF staining effectively distinguishes undifferentiated cells from the granulomonocytic maturation compartment.
Conclusions:
- Sensitive flow cytometric analysis of intracellular antigens is a powerful tool for hematopoiesis research and leukemia diagnostics.
- Intracellular MPO is a key marker for identifying early myeloid differentiation and diagnosing undifferentiated AML.
- The technology's potential extends to other intracellular markers like CD68 and various surface markers for comprehensive leukemia analysis.
Abstract:
New technology allows highly sensitive flow cytometric detection and quantitative analysis of intracellular antigens in normal and malignant hemopoietic cells. With this technology, the earliest stages of myeloid and lymphoid differentiation can easily and reliably be identified using antibodies directed against (pro-)myeloperoxidase/MPO, CD22 and CD3 antigens, respectively. Particularly for the analysis of undifferentiated acute myeloblastic leukemia (AML) cells, the immunological demonstration of intracellular MPO or its enzymatically inactive proforms is highly relevant, since other myeloid marker molecules such as CD33, CD13, or CDw65 are either not restricted to the granulomonocytic lineage or appear later in differentiation. By combining MPO staining with staining for lactoferrin (LF), undifferentiated cells can be distinguished from the granulomonocytic maturation compartment in bone marrow, since LF is selectively expressed from the myelocyte stage of differentiation onward. The list of informative intracellular antigens to be used in leukemia cell analysis will certainly expand in the near future. One candidate, intracellular CD68, has already been tested by us, and results are presented. Also dealt within this article are surface marker molecules not (as yet) widely used in leukemia cell analysis but with the potential to provide important additional information. Among them are the surface structures CD15, CD15s, CDw65, CD79a (MB-1), CD79b (B29), CD87 (uPA-R), and CD117 (c-kit).
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
05:24Two Flow Cytometric Approaches of NKG2D Ligand Surface Detection to Distinguish Stem Cells from Bulk Subpopulations in Acute Myeloid Leukemia
Published on: February 21, 2021
