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Published on: November 3, 2018
Immunophenotypic pattern of myeloid populations by flow cytometry analysis
Wojciech Gorczyca1, Zhong-Yi Sun, William Cronin
1Genzyme Genetics (New York Laboratory), New York, NY, USA.
Insights
This study details immunophenotypic markers for distinguishing benign and malignant myeloid cells, crucial for diagnosing conditions like acute myeloid leukemia (AML) and acute promyelocytic leukemia (APL). Understanding these cell characteristics aids in accurate myeloid disorder diagnosis.
Area of Science:
- Hematology
- Immunology
- Oncology
Background:
- Accurate immunophenotypic characterization is essential for differentiating benign from malignant myeloid populations.
- Distinguishing specific myeloid cell types, including eosinophils, dysplastic granulocytes, neoplastic promyelocytes, and monocytes, presents diagnostic challenges.
- Flow cytometry (FC) provides critical data for myeloid cell identification and classification.
Purpose of the Study:
- To present immunophenotypic characteristics of benign and malignant myeloid cells.
- To emphasize differential diagnostic strategies, particularly for eosinophils, dysplastic granulocytes, neoplastic promyelocytes, and monocytes.
- To describe distinct flow cytometry patterns observed in acute promyelocytic leukemia (APL).
Main Methods:
- Analysis of immunophenotypic markers using flow cytometry (FC).
- Detailed examination of cell scatter properties (SSC, FSC) and marker expression (CD antigens, HLA-DR).
- Comparison of marker profiles for various myeloid cell types and leukemic blasts.
Main Results:
- Established characteristic marker profiles for eosinophils (e.g., CD45 bright, CD11b+, CD14-) and mature monocytes (e.g., CD11b+, CD14+, CD64+).
- Defined immunophenotypes for acute myeloid leukemia (AML) blasts (e.g., CD34+, CD117+, CD13+, CD33+) and four distinct FC patterns in acute promyelocytic leukemia (APL).
- Highlighted differences between classical (hypergranular) and hypogranular (microgranular) APL variants based on SSC and marker coexpression (e.g., CD2, CD34).
Conclusions:
- Immunophenotyping provides a robust method for distinguishing benign and malignant myeloid cells.
- Specific FC patterns and marker combinations are crucial for diagnosing APL subtypes and differentiating from other myeloid disorders.
- Detailed phenotypic analysis aids in the accurate diagnosis and classification of myeloid malignancies.
Abstract:
We present our experience with immunophenotypic characteristics of benign and malignant myeloid populations, with emphasis on differential diagnosis especially between eosinophils, dysplastic granulocytes, neoplastic promyelocytes, and monocytes. Eosinophils are characterized by bright CD45, high side scatter (SSC), very low forward scatter (FSC), positive CD11b, CD11c, CD13, CD15, and CD33. They are negative for CD10, CD14, CD16, CD56, CD64, and HLA-DR. Mature monocytes are positive for CD11b, CD11c, CD13, CD14, CD33, and CD64, and may express CD2 and CD4. Blasts in acute myeloid leukemias (AML) with minimal differentiation have low SSC and moderate CD45 expression and are positive for CD34, CD117, CD13, HLA-DR, and CD33 and may be positive for TdT, CD4, and CD11c. In acute promyelocytic leukemia (APL), four FC patterns can be recognized. The majority of cases represented classical (hypergranular) APL and were characterized by high SSC, positive CD117, usually negative CD34, heterogeneous CD13, and bright CD33 (pattern 1). The second most common type, corresponding to hypogranular (microgranular) variant of APL, differed from classical APL by low SSC and frequent coexpression of CD2 and CD34 (pattern 2). Rare cases of APL (pattern3) showed mixture of neoplastic cells (SSC(low)/CD2(+)/CD13(+)/CD33(+)/CD34(+)/CD117(+)) and prominent population of benign granulocytes/maturing myeloid precursors (SSC(high)/CD10(+/-)/CD16(+/()/CD117(()). One case showed two APL populations, one with hypogranular and one with hypergranular characteristics (pattern 4). Detailed phenotypic characteristics of neoplastic monocytes and dysplastic granulocytes with their differential diagnosis are also presented.
