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Significantly improved PCR-based clonality testing in B-cell malignancies by use of multiple immunoglobulin gene targets. Report of the BIOMED-2 Concerted Action BHM4-CT98-3936

Leukemia volume 21, pages 207–214 (2007)Cite this article

Abstract

Polymerase chain reaction (PCR) assessment of clonal immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements is an important diagnostic tool in mature B-cell neoplasms. However, lack of standardized PCR protocols resulting in a high level of false negativity has hampered comparability of data in previous clonality studies. In order to address these problems, 22 European laboratories investigated the Ig/TCR rearrangement patterns as well as t(14;18) and t(11;14) translocations of 369 B-cell malignancies belonging to five WHO-defined entities using the standardized BIOMED-2 multiplex PCR tubes accompanied by international pathology panel review. B-cell clonality was detected by combined use of the IGH and IGK multiplex PCR assays in all 260 definitive cases of B-cell chronic lymphocytic leukemia (n=56), mantle cell lymphoma (n=54), marginal zone lymphoma (n=41) and follicular lymphoma (n=109). Two of 109 cases of diffuse large B-cell lymphoma showed no detectable clonal marker. The use of these techniques to assign cell lineage should be treated with caution as additional clonal TCR gene rearrangements were frequently detected in all disease categories. Our study indicates that the BIOMED-2 multiplex PCR assays provide a powerful strategy for clonality assessment in B-cell malignancies resulting in high Ig clonality detection rates particularly when IGH and IGK strategies are combined.

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Acknowledgements

We are grateful to the teams of the National Network Leaders and the technicians of the participating laboratories for their continuous support to make this European collaboration successful. We thank Elizabeth Macintyre (Paris), Els Moreau (Roeselare), Rita Fragoso (Lisbon), Patrocinio Algara (Toledo) and Lorenz Trumper (Gottingen). We acknowledge the help and commitment of Petra Chall, Till Lange (Kiel), Esther JM Schilder-Tol, Marjon J Clement (Amsterdam), Jelle Conradie, Klaas Kooistra (Groningen), Kathleen Louvaert (Roeselare), Margarethe C van Altena, Marian Verdijk (Nijmegen), Hans-Henning Müller, Hedwig Lammert (Berlin), Francoise Parmentier (Rouen), Christiane Copie-Bergman (Créteil), Francis Devez (Hôtel-Dieu, Paris), Christal Hercher (Pitié-Salpétière, Paris), Ben Swinson, Mike Short (Leeds), Mercedes Navarrete (Madrid), Sasala Wickramasinghe (Aberdeen) and Carole Charlot (Lyon). We also thank the secretaries of the coordinating center in Rotterdam for their continuous support, particularly Annella Boon and Bibi van Bodegom. We gratefully acknowledge Professor Martin Dyer, University of Leicester, for the kind gift of cell lines K231, Oz and SC1. We finally thank Ton Langerak for critical reading of this paper. This study was supported financially by the EU BIOMED-2 Grant BMH4-CT98-3936 as well as by national funding, including Revolving Fund 1995 project (JJMvD); Liga Portuguesa Contra o Cancro/NRS-1998/2001 (PG); Ministerio de Sanidad y Consumo (FIS G03/179), Spain (BM and RV); Deutsche Krebshilfe project No. 70-2405 (Ch.P).

Author information

Authors and Affiliations

  1. Haematological Malignancy Diagnostic Service, The General Infirmary at Leeds, Leeds, UK

    P A S Evans & G J Morgan

  2. II. Medical Department, University Hospital Schleswig-Holstein, Kiel, Germany

    Ch Pott

  3. Department of Pathology, Radboud University, Nijmegen Medical Center, Nijmegen, The Netherlands

    P J T A Groenen & J H J M van Krieken

  4. Department of Haematology, Centre Hospitalier Lyon-Sud, Lyon, France

    G Salles

  5. Department of Haematology, Hopital Pitié-Salpétière, Paris, France

    F Davi

  6. Department of Pathology, Centre Hospitalier Lyon-Sud, Lyon, France

    F Berger

  7. Molecular Pathology Program, CNIO, Madrid, Spain

    J F Garcia & R Villuendas

  8. Department of Pathology, Academic Medical Center, Amsterdam, The Netherlands

    S Pals & M Spaargaren

  9. Department of Pathology, University Medical Center Groningen, Groningen, The Netherlands

    Ph Kluin & E Schuuring

  10. Laboratory for Molecular Diagnostics, H. Hartziekenhuis, Roeselare, Belgium

    E Boone

  11. Department of Hematology, Hospital Universitario de Salamanca, Salamanca, Spain

    D González

  12. Department of Human Genetics, Centro Nacional de Investigaciones Oncológicas, Madrid, Spain

    B Martinez

  13. Lab. Biologia Molecular, Instituto Português de Oncologia, Lisbon, Portugal

    P Gameiro

  14. Department of Histopathology, Royal Free and UCL Medical School, London, UK

    T C Diss

  15. Department of Haematology, University of Wales College of Medicine, Cardiff, UK

    K Mills

  16. Molecular Diagnostics, Nottingham City Hospital NHS Trust, Nottingham, UK

    G I Carter

  17. Department of Medicine and Therapeutics, University of Aberdeen, Aberdeen, UK

    B J Milner

  18. Department of Pathology, Cambridge University, Addenbrooke's NHS Trust Hospital, Cambridge, UK

    D Pearson

  19. Institut für Pathologie, Campus Benjamin Franklin, Charité – Universitätsmedizin Berlin, Berlin, Germany

    M Hummel

  20. Department of Internal Medicine, Georg-August-University of Goettingen, Goettingen, Germany

    W Jung

  21. Department of Pathology, Luitpoldkrankenhaus, Würzburg, Germany

    M Ott

  22. Service d’Anatomie Pathologique, Hôpital Necker-Enfants Malades, Paris, France

    D Canioni

  23. Laboratoire d’Hematologie, Hôpital Necker-Enfants Malades, Paris, France

    K Beldjord

  24. Laboratoire de Genetique Oncologique, Centre Henri Becquerel, Rouen, France

    C Bastard

  25. Service d’Immunologie Biologique, Groupe Hospitalier Henri Mondor, AlbertChenevier AP-HP, Creteil, France

    M H Delfau-Larue

  26. Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands

    J J M van Dongen

  27. Department of Pathology, Hôtel-Dieu AP-HP, Paris, France

    T J Molina

  28. Serviço de Anatomia Patológica, Instituto Portugués de Oncologia, Lisbon, Portugal

    J Cabeçadas

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  1. P A S Evans
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  32. T J Molina
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Correspondence to J J M van Dongen.

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Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

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Evans, P., Pott, C., Groenen, P. et al. Significantly improved PCR-based clonality testing in B-cell malignancies by use of multiple immunoglobulin gene targets. Report of the BIOMED-2 Concerted Action BHM4-CT98-3936. Leukemia 21, 207–214 (2007). https://doi.org/10.1038/sj.leu.2404479

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