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Updated: Aug 14, 2026
Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
Published on: January 9, 2019
Immunohistochemistry in apparently normal bone marrow trephine specimens from patients with nodal follicular lymphoma
R Chetty1, G Echezarreta, M Comley
1University Department of Cellular Science, John Radcliffe Hospital, Oxford.
Insights
Immunohistochemistry can detect minimal follicular lymphoma in bone marrow, even when standard tests show none. Strong bcl-2 staining in bone marrow lymphoid aggregates indicates lymphoma in patients with nodal follicular lymphoma.
Area of Science:
- Hematopathology
- Oncology
- Immunohistochemistry
Background:
- Follicular lymphoma can involve the bone marrow with minimal or no morphological evidence.
- Distinguishing between benign and malignant lymphoid infiltrates in bone marrow is crucial.
Purpose of the Study:
- To assess the utility of immunohistochemistry in detecting minimal follicular lymphoma in bone marrow trephine specimens.
- To evaluate bcl-2 immunostaining for differentiating benign from malignant bone marrow infiltrates in nodal follicular lymphoma.
Main Methods:
- Bone marrow trephine specimens (n=27) were analyzed using immunohistochemistry.
- Antibodies used included anti-bcl-2, anti-CD79a, anti-CD3, and kappa/lambda light chains.
- Streptavidin biotin complex technique was employed for staining.
Main Results:
- Immunohistochemistry identified monotypic B cells in 5 of 27 cases initially negative for follicular lymphoma.
- These minimal infiltrates, diffuse or in mini-aggregates, were strongly bcl-2 positive.
- Reactive B lymphoid nodules showed weaker bcl-2 positivity with discernible negative cells.
Conclusions:
- Immunohistochemistry is valuable for detecting minimal follicular lymphoma in bone marrow.
- A limited antibody panel (anti-bcl-2, anti-CD79a, anti-CD3) is generally sufficient.
- Strongly bcl-2 positive lymphoid aggregates in bone marrow suggest lymphoma in patients with nodal follicular lymphoma.
Aim:
To establish the role of immunohistochemistry (using a limited panel of antibodies) in detecting minimal involvement by follicular lymphoma in routinely processed bone marrow trephine specimens, which show no obvious morphological (light microscopic) evidence of lymphoma; to determine whether bcl-2 immunostaining in bone marrow distinguishes between benign and malignant infiltrates in a patient with nodal follicular lymphoma.
Methods:
Twenty seven consecutively selected paraffin wax embedded, formalin fixed bone marrow trephine specimens were stained with the following antibodies: anti-bcl-2, anti-CD79a, anti-CD3, and kappa and lambda light chains, using the Streptavidin biotin complex technique.
Results:
Five of the 27 cases, which showed no evidence of involvement by follicular lymphoma on routine stains, showed monotypic B cells on immunohistochemistry. Two of the cases were diffuse, while the remaining three showed mini-aggregates around bony trabeculae. In all five cases the lymphomatous infiltrates were strongly bcl-2 positive. Reactive B lymphoid nodules did not show the same degree of bcl-2 positivity, and negative cells could be discerned within the reactive nodules.
Conclusions:
There is merit in studying so-called negative bone marrows immunohistochemically in order to detect minimal involvement by follicular lymphoma. A limited panel of antibodies including anti-bcl-2, anti-CD79a and anti-CD3 is usually adequate to accomplish this. Strongly bcl-2 positive lymphoid aggregates in the bone marrow of patients with nodal follicular lymphoma are indicative of lymphoma.
