Mantle cell lymphoma: improved diagnostics using a combined approach of immunohistochemistry and identification of t(11;14)(q13;q32) by polymerase chain reaction and fluorescence in situ hybridization

Abstract

Mantle cell lymphoma: improved diagnostics using a combined approach of immunohistochemistry and identification of t(11;14)(q13;q32) by polymerase chain reaction and fluorescence in situ hybridization Abstract Introduction: Mantle cell lymphoma (MCL) is a clinicopathological entity characterized by an aggressive clinical course, morphological features, and overexpression of cyclin D1 due to juxtaposition of the bcl-1 locus (and CCND1 gene coding for the cyclin D1) to the IgH gene. This phenomenon is caused by t(11;14)(q13;q32). The morphological diagnosis of MCL may pose difficulties. Ancillary methods are available to support the diagnosis. Patients and methods: We studied a group of 32 patients with MCL; 24 men and 8 women. The median age at the diagnosis was 64 years. We characterized the investigated group by histology, and to analyze the immunohistochemical (IHC) profile we used a panel of antibodies including anti-cyclin D1. Polymerase chain reaction (PCR) was used to detect the rearrangement of bcl-1/IgH in 26 cases (in 11 patients, the DNA was isolated from frozen tissues or from nucleated cells of bone-marrow aspirate or peripheral blood, in 15 patients we utilized paraffin-embedded material). Dual color fluorescence in situ hybridization (FISH) on interphase nuclei detecting the t(11;14)(q13;q32) was applied in all 32 cases. Results: Cyclin D1 IHC was positive in 29 of 30 cases tested (97%). In six, the result was weak and difficult to rely on to support the diagnosis. PCR revealed the fusion gene in 14 of the 26 cases (54%). The best yield was obtained from fresh and frozen samples (8 of 11 positive). Using FISH, we identified the translocation in all 32 patients, the findings being easily interpretable in 29 patients. In three cases, the intensity of red and green signals was weaker and difficult to read though the cohybridized signals were identified. The classical pattern of the translocation was observed in 26 patients, while in 3 we found variant patterns suggesting a loss of the V segment of the IgH gene (2x) and a shift in the breakpoint region at chromosome 11 (1x). Conclusion: The diagnosis of MCL should be supported by a complex laboratory approach. Interphase FISH seems a useful complementary method to morphology and IHC. It is applicable to various tissues and cells prepared as tissue imprints or histological sections.

CD5‐positive mucosa‐associated lymphoid tissue (MALT) lymphoma of ocular adnexal origin: Usefulness of fluorescence in situ hybridization for distinction between mantle cell lymphoma and MALT lymphoma

“…According to the combined studies of t(11;14) by molecular cytogenetic and immunohistochemical analysis in a series of MCL cases, the positivity of interphase FISH analysis (86–100%) is generally higher than that of DNA‐PCR for detection of bcl‐1/IgH rearrangement (37–54%), reverse transcription–polymerase chain reaction (RT‐PCR) for cyclin D1 transcript (77–89%) and immunohistochemistry for cyclin D1 protein expression (69–100%) 11,15,17,18,21,22 . Because the high sensitivity and applicability on paraffin‐embedded sections largely exceed those of other techniques, several investigators propose interphase FISH as the method of choice for determination of t(11;14) on routine pathological specimens of MCL 15,16 . In the present study, the number of positive cells carrying BCL1/IgH fusion signal in the positive control MCL case was 89 out of 200 (44.5%), while the present patients 1 and 2 had only two (1.0%) and three (1.5%), respectively.…”

“…3a). 15–17 The number of these positive nuclei was 89 out of 200 (44.5%). In contrast, the negative control had two separate BCL1 and IgH signals in the nuclei of non‐neoplastic lymphocytes, which is a typical pattern seen in non‐rearranged cells (data not shown).…”

“…Peripheral blood smears from healthy individuals were added as negative controls. Details of the procedure have been previously reported 15 . The probe is LSI IGH/CCND1 dual‐color, dual‐fusion translocation probe (Vysis, Downers Grove, IL, USA), which are designed to identify the locus 14q32 (IGH) and 11q13 (CCND1).…”

“…In an abnormal cell containing reciprocal t(11;14)(q13;q32), two colocalized orange/green fusion signals, with a single orange signal for intact allele of 11q13, and a single green signal for intact 14q13, are observed. Other variant patterns may be observed depending on the breakpoints involved in the translocation or other rearrangements 15 …”

First pathological report of a de novo CD5‐positive diffuse large B‐cell lymphoma patient presenting with Guillain–Barré syndrome‐like neuropathy due to neurolymphomatosis

“…These cyclin D1-positive cells were considered to be enlarged endothelial cells or histiocytes. 4 Translocation at t(11;14)(q13;q32) which is present in almost all cases of mantle cell lymphoma, 5 was not detected by fluorescence in situ hybridization. The patient was therefore diagnosed with de novo CD5-positive DLBCL.…”