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Biomed-2

Powerful strategy for polymerase chain reaction-based clonality assessment in T-cell malignancies Report of the BIOMED-2 Concerted Action BHM4 CT98-3936

Abstract

Polymerase chain reaction (PCR) assessment of clonal T-cell receptor (TCR) and immunoglobulin (Ig) gene rearrangements is an important diagnostic tool in mature T-cell neoplasms. However, lack of standardized primers and PCR protocols has hampered comparability of data in previous clonality studies. To obtain reference values for Ig/TCR rearrangement patterns, 19 European laboratories investigated 188 T-cell malignancies belonging to five World Health Organization-defined entities. The TCR/Ig spectrum of each sample was analyzed in duplicate in two different laboratories using the standardized BIOMED-2 PCR multiplex tubes accompanied by international pathology panel review. TCR clonality was detected in 99% (143/145) of all definite cases of T-cell prolymphocytic leukemia, T-cell large granular lymphocytic leukemia, peripheral T-cell lymphoma (unspecified) and angioimmunoblastic T-cell lymphoma (AILT), whereas nine of 43 anaplastic large cell lymphomas did not show clonal TCR rearrangements. Combined use of TCRB and TCRG genes revealed two or more clonal signals in 95% of all TCR clonal cases. Ig clonality was mostly restricted to AILT. Our study indicates that the BIOMED-2 multiplex PCR tubes provide a powerful strategy for clonality assessment in T-cell malignancies assisting the firm diagnosis of T-cell neoplasms. The detected TCR gene rearrangements can also be used as PCR targets for monitoring of minimal residual disease.

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Acknowledgements

We are grateful to the teams of the National Network Leaders and the technicians for their continuous support to make this European collaboration successful. We particularly thank Frauke Hemken (Kiel), Ana Balanzategui, David González (Salamanca), Rita Fragoso (Lisboa), Gabi Krull (Münster), Monique ECM Oud, Ellen J van Gastel-Mol, Ingrid LM Wolvers-Tettero, Brenda Verhaaf (Rotterdam), Esther JM Schilder-Tol, Marjon J Clement (Amsterdam), Elke Boone, Kathleen Louvaert (Roeselare), Margarethe C van Altena, Marian Verdijk (Nijmegen), Patrícia Pontes (Porto), Hans-Henning Müller, Hedwig Lammert (Berlin), Francoise Parmentier (Rouen), Christiane Copie-Bergman (Créteil), Maryse Magen, Véronique Minard-Colin (Hôpital Necker-Enfants Malades, Paris) and Francis Devez (Hotel-Dieu de Paris, Paris) for technical assistance and Louise Lavender, Elizabeth Hodges and John Smith (Southampton) for their continuous support. We also thank the secretaries of the coordinating center in Rotterdam, particularly Annella Boon and Bibi van Bodegom. 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 and AWL), Deutsche Krebshilfe Grant Kn-70354-2 (MK), Fondation Contre la Leucémie of the Fondation de France (KB), Liga Portuguesa Contra o Cancro/NRS-1998/2001 (PG), Grants from the ‘Fondo de Investigaciones Sanitarias del Ministerio de Sanidad y Consumo’ number FIS-02/1244 (AO) and FIS-PI02/0905 (RGS) and Leukaemia Research Fund Grant 0141 (HW).

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Brüggemann, M., White, H., Gaulard, P. et al. Powerful strategy for polymerase chain reaction-based clonality assessment in T-cell malignancies Report of the BIOMED-2 Concerted Action BHM4 CT98-3936. Leukemia 21, 215–221 (2007). https://doi.org/10.1038/sj.leu.2404481

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