What is the immunophenotype of lymphoma cells?

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Immunophenotype of Lymphoma Cells

The immunophenotype of lymphoma cells varies by lymphoma subtype and is established through flow cytometry or immunohistochemistry using specific panels of cell surface and intracellular markers that define B-cell, T-cell, or NK-cell lineage and differentiate between lymphoma types. 1

Essential Immunophenotyping Approach

Core Diagnostic Panels

For initial lymphoma diagnosis, two primary methods are used:

  • Flow cytometry panel should include: kappa/lambda light chains, CD19, CD20, CD5, CD23, and CD10 1
  • Immunohistochemistry panel should include: CD20, CD3, CD5, CD10, BCL2, BCL6, cyclin D1, and CD21 or CD23 1

The NCCN guidelines emphasize that adequate immunophenotyping is essential (not optional) to establish diagnosis, and flow cytometry of peripheral blood is often sufficient for leukemic presentations while tissue biopsy with immunohistochemistry is preferred for nodal disease 1

Specific Lymphoma Immunophenotypes

B-Cell Lymphomas

Follicular Lymphoma (Grade 1-2):

  • Typical pattern: CD10+, BCL2+, CD20+, BCL6+, CD23+/-, CD43-, CD5-, cyclin D1- 1
  • Rare cases may be CD10- or BCL2- 1

Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma (CLL/SLL):

  • Characteristic pattern: CD5+, CD10-, CD19+, CD20 dim, surface immunoglobulin dim, CD23+, CD43+/-, cyclin D1- 1
  • Critical distinction: Absence of cyclin D1 expression is essential to differentiate from mantle cell lymphoma 1
  • FISH for t(11;14) should be performed if flow cytometry alone is used to rule out mantle cell lymphoma 1

Mantle Cell Lymphoma:

  • Pattern: CD5+, CD20+, cyclin D1+, CD23- (typically) 1
  • Distinguished from CLL/SLL by cyclin D1 positivity 1

Burkitt Lymphoma:

  • Pattern: surface Ig+, CD10+, CD20+, TdT-, Ki67+ (approaching 100%), BCL2-, BCL6+, with MYC rearrangement 1

T-Cell and Precursor Lymphomas

B-Lymphoblastic Leukemia/Lymphoma:

  • Pre-pre-B (pro-B): TdT+, CD19+/CD22+/CD79a+, CD10-, surface Ig- 1
  • Common B-cell: CD10+, TdT+, CD19+/CD22+/CD79a+ 1
  • Pre-B: cytoplasmic Ig+, CD10+, CD19+/CD22+/CD79a+ 1
  • Mature B-cell: surface Ig+, clonal kappa or lambda, TdT- 1
  • CD20 expressed in approximately 50% of B-lineage cases, >80% in mature B-cell cases 1

T-Lymphoblastic Leukemia/Lymphoma:

  • Pattern: cytoplasmic CD3+ or surface CD3+, CD1a/CD2/CD5/CD7 (variable), TdT+ 1
  • Surface Ig-, CD10-, CD19/20-, CD4/8+/+ (variable) 1
  • Early precursor T-cell subtype: CD1a-, CD8-, CD5 weak (<75% positive), with ≥1 myeloid/stem cell marker on ≥25% of blasts 1

Technical Considerations

Flow Cytometry vs. Immunohistochemistry

Flow cytometry advantages:

  • Rapid diagnosis with multiparameter single-cell evaluation 2
  • Superior for detecting clonality in both B-cells (light chain restriction) and T-cells 2
  • Can evaluate surface, cytoplasmic, and nuclear antigens simultaneously 2
  • Essential for minimal residual disease detection 3

Immunohistochemistry advantages:

  • Preserves tissue architecture 1
  • Required when flow cytometry is non-diagnostic 1
  • Necessary for certain markers like Ki-67 proliferation index 1

Critical Diagnostic Pitfalls

Common errors to avoid:

  • Do not rely on FNA or core biopsy alone for initial lymphoma diagnosis; excisional biopsy is preferred, though combined core/FNA with comprehensive ancillary studies may be sufficient when lymph nodes are inaccessible 1
  • Always exclude mantle cell lymphoma when evaluating CD5+ B-cell lymphomas by confirming cyclin D1 negativity or absence of t(11;14) 1
  • Recognize CD23 overlap: One-third of follicular lymphomas coexpress CD23 and CD10, which can cause confusion with CLL/SLL 1
  • Post-therapy changes: Anti-CD19, anti-CD20, and anti-CD38 therapies can alter immunophenotype and complicate interpretation 4

Additional Useful Markers

Under certain circumstances, add:

  • Molecular analysis for antigen receptor gene rearrangements and BCL2 rearrangement 1
  • Cytogenetics or FISH for t(14;18), t(8;14), BCL6 rearrangements 1
  • Ki-67 proliferation index (>30% in follicular lymphoma may indicate aggressive behavior) 1
  • EBER-ISH for Epstein-Barr virus in appropriate clinical contexts 1

The immunophenotype must be interpreted in conjunction with morphology, clinical presentation, and genetic findings to establish accurate lymphoma classification and guide treatment decisions. 1, 2

References

1
Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

2
Research

Flow cytometric analysis of lymphomas: current status and usefulness.

Archives of pathology & laboratory medicine, 2006

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This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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