Catalyzed Signal Amplification for Cyclin D1 Detection in Mantle Cell Lymphoma

Abstract

Mantle cell lymphoma is characterized by a t(11; 14)(q13;q32) translocation resulting in cyclin D1 protein overexpression. Immunohistochemical detection of the latter, therefore, is a useful marker for the diagnosis of mantle cell lymphoma. Nevertheless, interpretation of results is often hampered by the weak immunoreactivity obtained with routine detection techniques. This problem can be overcome by resorting to highly sensitive catalyzed signal amplification methods based on peroxidasecatalyzed deposition of a biotinylated phenolic compound. The present study compares the results obtained with catalyzed signal amplification, labeled streptavidin biotin, and dextran polymeric conjugate (EnVision؉) techniques in cyclin D1 demonstration in mantle cell lymphoma. The study was performed on formalin-fixed, paraffinembedded archival tissue from 20 mantle cell lymphoma cases. Ten cases of small lymphocytic lymphoma and 10 instances of follicular center cell lymphoma were used as controls. Mantle cell lymphoma, a malignant proliferation of small or medium-sized B lymphocytes that are the neoplastic counterpart of normal lymphoid follicle mantle cells, adopts a variety of growth patterns. This neoplasm is associated with a median survival of 3-5 years and lack of cure in the vast majority of patients. In fact, the 5-year overall survival of mantle cell lymphoma patients (30%) is among the worst for the various lymphoma types (1-7). Distinguishing this more aggressive type of B-cell lymphoma from others (marginal zone, small lymphocytic, and follicular lymphomas) can be very difficult.The most useful mantle cell lymphoma marker is cyclin D1, a cell cycle regulator that stimulates transition from G1 phase to S phase by cooperating with cdk4-6 in Rb phosphorylation and E2F release. Normally, lymphocytes do not express cyclin D1 or produce it in very low, immunohistochemically undetectable levels. In contrast, mantle cell lymphoma lymphocytes show immunohistochemically demonstrable cyclin D1 overexpression as a consequence of a t(11;14)(q13;q32) translocation involving the bcl-1 region of chromosome 11 and the immunoglobulin heavy chain gene of chromosome 14 (8 -10). This translocation is shown by cytogenetics or Southern blotting techniques in 70 -75% of mantle cell lymphoma cases and by fluorescence in situ hybridization (FISH) studies in virtually all mantle cell lymphoma cases (11). As for immuno-

Immunohistological recognition of cyclin D1 expression by non-lymphoid cells among lymphoid neoplastic cells

“…Banno and Co‐workers showed a correlation of Cyclin D1 mRNA overexpression with the immunohistochemical demonstration of Cyclin D1 in Mantle cell lymphoma and myeloma cell lines with t (q) translocation and breast carcinoma with 11q13 amplification . Studies on Mantle cell lymphoma cases with typical histology, immunophenotype and t (q13; q32) translocation consistently revealed the same cellular localisation of immunoreactivity .…”

“…The highest frequency, up to 20% of cases, is seen in multiple myeloma . Other lymphoproliferative disorders include lymphoplasmacytic lymphoma (LPL) , splenic marginal zone lymphoma , chronic cell lymphoma (CLL) , hairy cell leukaemia , ALK (−) Anaplastic large cell lymphoma and even classical Hodgkin lymphoma . The reactivity in the latter cases was confined to Reed‐Sternberg cells.…”

Immunolabeling recovery in archival, post-mortem, human brain tissue using modified antigen retrieval and the catalyzed signal amplification system

“…Past studies have indeed suggested that formic acid might be a more effective in breaking formalin-induced covalent bonds between proteins and nucleic acids than heat treatment, particularly in immunohistochemical studies using tau, amyloid, prion protein markers (Cammarata et al, 1990;Pikkarainen et al, 2010). Past studies have demonstrated that the CSA system could detect target proteins, even at low level (Hashizume et al, 2001;Barranco et al, 2007). This is because the system effectively amplifies signals by catalyzing the oxidation and precipitation of the fluorescein-labeled phenol compounds (fluorescyl-tyramide) onto sections, which are then detected using peroxidase conjugated anti-fluorescein antibodies.…”

“…Our present study aims to extend these past studies by examining the effect of long-term formalin fixation on the immunoreactivities of 20 antibodies, 10 of which have not been investigated in past studies using post-mortem, human brain tissue (anti-von Willebrand factor, vWF, anti-caveolin, anti-claudin 5, anti-P-glycoprotein (Pgp, JSB1, C219, C494), anti-breast cancer resistance protein, BCRP, anti-multidrug resistance related protein 1, MRP1, anti-human leukocyte antigen, HLA-DR, anti-connexin 43, Cx43). In view of the study by Pikkarainen et al (2010), we also investigate for the first time whether formic acid pre-treatment in combination with antigen retrieval and/or the catalyzed signal amplification (CSA) system (which allows the detection of protein even at low level; Hashizume et al, 2001;Barranco et al, 2007), is effective in recovering the immunoreactivities of formalin-sensitive antibodies.…”

Gastrointestinal Involvement in Mantle Cell Lymphoma: A Prospective Clinic, Endoscopic, and Pathologic Study

“…As positive controls, an MCL case was used for cyclin D1 and a normal tonsil for the remaining antibodies. Catalyzed signal amplification, as previously described, 3 was applied to cases with negative or inconclusive cyclin D1 results. Only cyclin D1 nuclear staining was accepted as a genuinely positive result.…”