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Immunogold labeling for the diagnosis of leukemia by transmission and scanning electron microscopy
E de Harven1, D Soligo, H Christensen
1Department of Pathology, University of Toronto, Canada.
Insights
Immuno-electron microscopy aids leukemia diagnosis in adults. CD103 confirms hairy cell leukemia, but CD61 findings suggest circulating megakaryocythemia, not true megakaryoblastic leukemia (M7).
Area of Science:
- Hematology
- Oncology
- Immunology
Background:
- Diagnosing leukemia in adults can be challenging, especially when bone marrow sampling is difficult.
- Peripheral blood leukocyte (PBL) analysis offers an alternative diagnostic avenue.
Purpose of the Study:
- To evaluate the utility of immuno-electron microscopy in diagnosing leukemia subtypes, specifically hairy cell leukemia and megakaryoblastic leukemia (M7).
- To differentiate between true megakaryoblastic leukemia and other conditions with megakaryocytic features.
Main Methods:
- Indirect immunogold labeling using B-ly7 monoclonal antibody (CD103) for hairy cell leukemia.
- Immunogold labeling for GpIIIa platelet glycoprotein (CD61) in suspected megakaryoblastic leukemia cases.
- Electron microscopy examination of peripheral blood leukocytes.
Main Results:
- CD103 labeling confirmed hairy cell leukemia diagnoses.
- CD61-positive cells in suspected M7 cases exhibited extensive megakaryopoietic differentiation, including alpha granules and demarcation membranes.
- These findings indicated circulating megakaryocythemia rather than true megakaryoblastic leukemia in most cases.
Conclusions:
- Immuno-electron microscopy is valuable for leukemia diagnosis, particularly when bone marrow access is limited.
- CD61 expression in peripheral blood cells may indicate megakaryocythemia, necessitating careful re-evaluation of megakaryoblastic leukemia (M7) diagnoses.
- Distinguishing between M7 and other myeloproliferative disorders with megakaryocytic features is crucial for accurate patient management.
Abstract:
For the cell type diagnosis of leukemia in adult patients, particularly when the sampling of bone marrow is difficult, the study of peripheral blood leukocytes (PBLs) by immuno-electron microscopy provides significant information, as illustrated here in two cases of hairy cell leukemia and seven cases tentatively identified as megakaryoblastic leukemia (M7). Indirect immunogold labeling with the B-ly7 monoclonal antibody (CD103) proved valuable in confirming the diagnosis of hairy cell leukemia. Immunogold labeling for the GpIIIa platelet glycoprotein (CD61) was used in cases where the light microscopy of blood films revealed possible megakaryoblastic leukemia. Under the electron microscope, however, the CD61 positive cells showed, in almost all cases, a wide spectrum of megakaryopoietic differentiation which made the diagnosis of M7 questionable. Most of the CD61 positive cells featured cytoplasmic differentiation markers such as alpha granules and demarcation membranes, further confirming the presence of circulating megakaryocythemia, a phenomenon described many years ago in various myeloproliferative disorders. It is suggested, therefore, that many of these cases should not be identified as true megakaryoblastic leukemias.
