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Updated: Aug 8, 2026

Flow 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
[Analysis of cytoplasmic antigens in acute leukemia by flow cytometry]
1Department of Pediatrics, Aichi Medical University.
Insights
Cytoplasmic antigen analysis using flow cytometry aids in classifying acute leukemia subtypes. Detecting CD3, CD22, mu-chain, and myeloperoxidase (MPO) antigens accurately defines cell lineage in acute lymphoblastic leukemia (ALL) and acute non-lymphoblastic leukemia (ANLL).
Area of Science:
- Immunophenotyping
- Hematologic Malignancies
- Flow Cytometry
Context:
- Accurate classification of acute leukemia is crucial for effective treatment strategies.
- Distinguishing between acute lymphoblastic leukemia (ALL) and acute non-lymphoblastic leukemia (ANLL) relies on precise cell lineage determination.
- Traditional cytochemical staining may have limitations in detecting certain antigens.
Purpose:
- To evaluate the utility of cytoplasmic antigen expression analysis via flow cytometry in classifying 77 cases of acute leukemia.
- To investigate the expression of CD3, CD22, cytoplasmic mu-heavy chain, and myeloperoxidase (MPO) antigens.
- To determine if these cytoplasmic markers can reliably define cell lineage and differentiate ALL and ANLL subtypes.
Summary:
- Cytoplasmic CD22 was detected in most non-T-ALL cases, including some unclassified ALL with absent surface CD22.
- Cytoplasmic mu-heavy chain identified Pre-B ALL, while cytoplasmic CD3 was present in T-ALL cases lacking surface CD3.
- Cytoplasmic MPO was prevalent in ANLL (20/21 cases), with flow cytometry detecting MPO in cases negative by conventional staining; notably, MPO was also found in a subset of T-ALL cases.
Impact:
- The study demonstrates that analyzing cytoplasmic antigens (CD3, CD22, mu-chain, MPO) is highly effective for leukemia cell lineage definition and classification.
- Findings support the use of flow cytometry for detecting cytoplasmic antigens, improving diagnostic accuracy for ALL and ANLL.
- Further research is warranted to clarify the significance of cytoplasmic MPO expression in T-ALL.
Abstract:
The expression of cytoplasmic antigens in 77 cases of acute leukemia were analyzed by flow cytometry using the following monoclonal antibodies: CD3, CD22, anti-myeloperoxidase (MPO-7) and anti-mu-heavy chain. CD22 antigen was detected in the cytoplasm of all non-T-ALL patients excluding one not-tested patient. In two patients with unclassified ALL, surface CD22 antigen was not expressed but cytoplasmic CD22 antigen was strongly expressed. Three out of 9 patients with common ALL were cytoplasmic mu-heavy chain-positive, so these patients were diagnosed as Pre-B ALL. In four out of 8 patients with T-ALL, CD3 antigen was not expressed on the cell surface membrane. However all of T-ALL patients excluding one non-tested patient were cytoplasmic CD3-positive. The cytoplasmic expression of myeloperoxidase antigen was detected in twenty out of 21 patients with acute non-lymphoblastic leukemia (ANLL). One megakaryocytic leukemia patient was MPO-negative. In two ANLL patients, the percentage of MPO for conventional cytochemical staining was undetectable or low, but MPO antigens were positive (77% and 70%) for flow cytometric analysis. All of 46 non-T ALL patients were cytoplasmic MPO-negative, however 4 out of 10 T-ALL patients were cytoplasmic MPO-positive. The study proved that the analysis of cytoplasmic CD3, CD22, mu-chain and MPO antigens were very useful to define the cell lineage of leukemia and to classify ALL and ANLL. It is necessary to study further whether the expression of MPO in the cytoplasm of T-ALL was non-specific reaction or whether MPO precursors are expressed in the cytoplasm of T-ALL.
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