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Updated: May 22, 2026
Flow Cytometric Characterization of Murine B Cell Development
Published on: January 22, 2021
Immunophenotyping of chronic B-cell neoplasms: flow cytometry versus immunohistochemistry
Afaf Abdel-Aziz Abdel-Ghafar1, Manal Ahmed Shams El Din El Telbany, Hanan Mohamed Mahmoud
1Clinical and Chemical Pathology, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
Insights
Immunophenotyping by flow cytometry and immunohistochemistry are equally effective for diagnosing chronic lymphocytic leukemia (CLL). Immunohistochemistry may offer greater sensitivity for detecting bone marrow infiltration in non-Hodgkin lymphoma (NHL) and hairy cell leukemia (HCL).
Area of Science:
- Hematology
- Oncology
- Pathology
Background:
- Differentiating benign from malignant lymphoproliferative disorders (LPDs) presents diagnostic challenges.
- Immunophenotyping (IPT) using flow cytometry or immunohistochemistry (IHC) is crucial for accurate LPD diagnosis.
Purpose of the Study:
- To compare the diagnostic efficacy of flow cytometry and IHC in chronic B-cell neoplasms.
- To evaluate the sensitivity of IHC versus flow cytometry in detecting bone marrow infiltration in specific LPDs.
Main Methods:
- Flow cytometry analysis of bone marrow samples using a comprehensive LPD panel on 35 patients.
- Immunohistochemistry staining of bone marrow trephine biopsy specimens for key markers (CD20, CD5, CD23, Bcl-2, Bcl-6, kappa, lambda).
Main Results:
- Both IHC and flow cytometry demonstrated equal effectiveness in diagnosing chronic lymphocytic leukemia (CLL).
- IHC showed higher sensitivity than flow cytometry in detecting bone marrow infiltration in non-Hodgkin lymphoma (NHL) and hairy cell leukemia (HCL).
- Specific sensitivities for IHC were reported for CD20, CD5, CD23, and light chains in CLL and NHL.
Conclusions:
- Flow cytometry and IHC are comparable diagnostic tools for CLL.
- IHC may be superior to flow cytometry for assessing bone marrow involvement in NHL and HCL.
- Immunophenotyping remains a cornerstone in the diagnosis and classification of lymphoproliferative disorders.
Abstract:
Morphological differentiation between benign and malignant lymphoproliferative disorders (LPDs) can be challenging. Immunophenotyping (IPT) by either technique, flow cytometry or immunohistochemistry (IHC), is an important step in solving such difficulty. Thirty-five newly diagnosed patients with chronic B-cell neoplasms (11 chronic lymphocytic leukemia, 22 non Hodgkin lymphoma and 2 hairy cell leukemia) were included in this study with age range from 20 to 70 years. Monoclonal antibodies surface expression using lymphoproliferative disorders panel (CD45, CD19, CD5, CD10, CD11c, CD20, CD22, CD23, CD38, CD79b, FMC7, CD103, CD25, kappa and lambda light chains) by flow cytometry was done on bone marrow samples. CD20, CD5, CD23, Bcl-2, Bcl-6, kappa and lambda light chain immunostaining were performed on fixed bone marrow trephine biopsy specimen. The sensitivity of IHC was 81.8% in chronic lymphocytic leukemia (CLL) and 100% in non Hodgkin lymphoma (NHL) as regards CD20, 100% in both groups as regards CD5, 46% in CLL and 66.7% in NHL as regards CD23, 33.3% in CLL and 50% in NHL as regards kappa chain, 20% in CLL and 33.3% in NHL as regards lambda chain. We found that IHC and flow cytometry are equally effective in diagnosing CLL; however, IHC might be slightly more sensitive than flow cytometry in detecting bone marrow infiltration in NHL and hairy cell leukemia (HCL).
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