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Immunoglobulin light chain transcript detection by ultrasensitive RNA in situ hybridization for B-cell lymphoma diagnosis

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A Correction to this article was published on 16 November 2023

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Abstract

Evaluation of B-cell clonality can be challenging in the interpretation of lymphoid infiltrates on tissue sections. Clonality testing based on IG gene rearrangements analysis by PCR (IG-PCR) is the gold standard. Alternatively, B-cell clonality can be assessed by the recognition of immunoglobulin light chain (IgLC) restriction, by immunohistochemistry (IHC), chromogenic in situ hybridization (ISH) or flow cytometry (FC). IG-PCR requires molecular facilities, and FC requires cell suspensions, both not widely available in routine pathology units. This study evaluates the performance of B-cell clonality detection by IgLC-RNAscope® (RNAsc) in a group of 216 formalin-fixed, paraffin-embedded samples including 185 non-Hodgkin B-cell lymphomas, 11 Hodgkin lymphomas (HL) and 20 reactive samples. IgLC-RNAsc, performed in parallel with FC in 53 cases, demonstrated better performances (93% vs 83%), particularly in diffuse large B-cell lymphoma (98% vs 71%) and follicular lymphoma (93% vs 83%) diagnosis. IgLC-RNAsc was also superior to IHC and ISH especially in samples with limited tumor cell content, where IG-PCR was not informative. Performed for the first time on mediastinal lymphomas, IgLC-RNAsc identified monotypic IgLC transcripts in 69% of primary mediastinal large B-cell lymphoma (PMBCL) and 67% of mediastinal gray zone lymphomas (MGZL). IGK/L double-negative cells were detected in 1 PMBCL, 2 MGZL, and all classical HL, while monotypic IgLC expression appeared to be a hallmark in nodular lymphocyte-predominant HL. IgLC-RNAsc demonstrates to be a powerful tool in B-cell lymphoma diagnosis, above all in challenging cases with limited tumor cell content, ensuring in situ investigations on mechanisms of Ig regulation across lymphoma entities.

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Data Availability

The data that support the findings of this study are available on request from the corresponding author (LL).

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Funding

Economical support was provided by the Italian Ministry of University to FF (PRIN 2017ZXT5WR). CS was supported by the Italian Association for Cancer Research (AIRC; IG grant no. 23747). ZS activity was supported by Fondazione Beretta (Brescia, Italy).

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Authors and Affiliations

  1. Pathology Unit, Department of Molecular and Translational Medicine-DMMT, University of Brescia, Brescia, Italy

    Luisa Lorenzi, Silvia Lonardi, Michela Bonezzi, Arianna Valzelli, Piera Balzarini, William Vermi & Fabio Facchetti

  2. Pathology Unit, ASST Spedali Civili Di Brescia, Brescia, Italy

    Luisa Lorenzi, Stefania Zini, Mattia Bugatti, Flavia Melotti, Mattia Facchetti, Iacopo Ghini, Vincenzo Villanacci, Piera Balzarini, William Vermi & Fabio Facchetti

  3. Pathology Department, Surgical Pathology and Cytopathology Unit, Department of Medicine-DIMED, University of Padua School of Medicine, Padua, Italy

    Marco Pizzi & Angelo Paolo Dei Tos

  4. Flow Cytometry Unit, Clinical Chemistry Laboratory, ASST Spedali Civili Di Brescia, Brescia, Italy

    Viviana Giustini, Anna Galvagni & Marco Chiarini

  5. IFOM-ETS-The AIRC Institute of Molecular Oncology, Milan, Italy

    Stefano Casola

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  1. Luisa Lorenzi
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Contributions

FF and LL conceived and designed the study; LL, LS, BM, ZS, BuM, VA, BP, GV, GA, CM, and WV performed the experiments; MF, FM, GI, VV, PM, and APDT contributed with critical discussion on samples diagnosis; LL, BM, and LS analyzed the data; LL, CS, and FF contributed to interpretation of the results. All authors read and approved the manuscript.

Corresponding author

Correspondence to Luisa Lorenzi.

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Ethical issues

Experiments included in this study on human tissue were approved by the Local Ethical Committee (study FF-BCR2, code NP 3719).

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The authors declare no competing interests.

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The original online version of this article was revised:The presentation authors' given and family names has been corrected.

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Lorenzi, L., Lonardi, S., Bonezzi, M. et al. Immunoglobulin light chain transcript detection by ultrasensitive RNA in situ hybridization for B-cell lymphoma diagnosis. Virchows Arch (2023). https://doi.org/10.1007/s00428-023-03682-8

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  • DOI: https://doi.org/10.1007/s00428-023-03682-8

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