Original Article
Leukemia (2007) 21, 230–237. doi:10.1038/sj.leu.2404486; published online 14 December 2006
Human T-cell lines with well-defined T-cell receptor gene rearrangements as controls for the BIOMED-2 multiplex polymerase chain reaction tubes
Funding sources: none.
Y Sandberg1, B Verhaaf1, E J van Gastel-Mol1, I L M Wolvers-Tettero1, J de Vos1, R A F MacLeod2, J G Noordzij1, W A Dik1, J J M van Dongen1 and A W Langerak1
- 1Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands
- 2DSMZ – German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
Correspondence: Professor Dr JJM van Dongen, Department of Immunology, Erasmus MC, University Medical Center, Dr Molewaterplein 50, 3015 GE Rotterdam, The Netherlands. E-mail: b.van.bodegom@erasmusmc.nl
Received 15 February 2006; Revised 24 May 2006; Accepted 5 June 2006; Published online 14 December 2006.
Abstract
The BIOMED-2 multiplex polymerase chain reaction (PCR) tubes for analysis of immunoglobulin and T-cell receptor (TCR) gene rearrangements have recently been introduced as a reliable and easy tool for clonality diagnostics in suspected lymphoproliferations. Quality and performance assessment of PCR-based clonality diagnostics is generally performed using human leukemia/lymphoma cell lines as controls. We evaluated the utility of 30 well-defined human T-cell lines for quality performance testing of the BIOMED-2 PCR primers and protocols. The PCR analyses of the TCR loci were backed up by Southern blot analysis. The clonal TCRB, TCRG and TCRD gene rearrangements were analyzed for gene segment usage and for the size and composition of their junctional regions. In 29 out of 30 cell lines, unique clonal TCR gene rearrangements could be easily detected. Besides their usefulness in molecular clonality diagnostics, these cell lines can now be authenticated based on their TCR gene rearrangement profile. This enables their correct use in molecular clonality diagnostics and in other cancer research studies.
Keywords:
BIOMED-2, TCR, T-cell lines, clonality, TCRB, TCRG, TCRD
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