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Updated: Aug 24, 2026

Multiplex Cyclic Fluorescent Immunohistochemistry
Published on: January 26, 2024
Consistent immunostaining for cyclin D1 can be achieved on a routine basis using a newly available rabbit monoclonal
Wah Cheuk1, Kathy O Y Wong, Cesar S C Wong
1Department of Pathology, Queen Elizabeth Hospital, Kowloon, Hong Kong, SAR China. cheuk_wah@hotmail.com
Insights
A new rabbit monoclonal antibody (MAb) SP4 significantly improves cyclin D1 immunostaining consistency for diagnosing mantle cell lymphoma (MCL). This enhanced antibody performance aids in differentiating MCL from other B-cell lymphomas.
Area of Science:
- Immunohistochemistry
- Hematopathology
- Oncology
Background:
- Cyclin D1 immunohistochemistry is challenging for diagnosing mantle cell lymphoma (MCL).
- Rabbit monoclonal antibodies (MAbs) offer potential for improved affinity and specificity.
Purpose of the Study:
- To evaluate a new rabbit MAb (SP4) against cyclin D1 for consistent immunostaining.
- To assess its utility in diagnosing MCL and differentiating it from other B-cell lymphomas.
Main Methods:
- Immunohistochemical staining of 150 lymphoproliferative lesions using rabbit MAb SP4.
- Comparison with mouse MAb DCS-6 and assessment of staining scores.
- Utilized formalin-fixed, paraffin-embedded tissues and an automated immunostainer.
Main Results:
- Rabbit MAb SP4 demonstrated superior performance with higher median staining scores in MCL cases compared to mouse MAb.
- All MCL cases expressed cyclin D1.
- Rare cases of B-cell chronic lymphocytic leukemia (B-CLL), multiple myeloma, and hairy cell leukemia also showed cyclin D1 positivity.
Conclusions:
- The rabbit MAb SP4 provides consistent and reliable cyclin D1 immunostaining, beneficial for MCL diagnosis.
- Cyclin D1 immunohistochemistry remains valuable for differential diagnosis, with awareness of potential positivity in other B-cell malignancies like B-CLL.
Abstract:
Rabbit monoclonal antibody (MAb), which has become available only recently, theoretically combines the advantage of the high affinity attributable to its rabbit origin and the high specificity due to its monoclonal nature. Since immunohistochemical demonstration of cyclin D1 is notoriously difficult, this study aims to assess whether a newly available rabbit MAb against cyclin D1 (SP4) can improve the consistency of immunostaining, especially for the diagnosis of mantle cell lymphoma (MCL). A total of 150 cases of lymphoproliferative lesions, including 30 cases of MCL, histologic mimickers of MCL, and various types of lymphomas and leukemias, were studied. Immunostaining was performed on formalin-fixed, paraffin-embedded tissue sections using a labeled streptavidin-biotin peroxidase system in an automated immunostainer. All cases of MCL expressed cyclin D1, with a higher median staining score (8 out of a maximum of 12) compared with mouse MAb DCS-6 (score 4). In addition, 2 of 15 cases of B-cell chronic lymphocytic leukemia (B-CLL), 3 of 12 cases of multiple myeloma, and 2 of 5 cases of hairy cell leukemia were also positive. Comparable staining results could also be achieved by an optimized manual staining protocol. This study thus confirms the superior performance of the rabbit MAb SP4, which should permit consistent immunostaining for cyclin D1 to be readily achieved. The value of cyclin D1 immunohistochemistry in the differential diagnosis of MCL from other low-grade B-cell lymphomas is also affirmed, but with the caveat that rare cases of B-CLL can also be cyclin D1 positive.
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