title: Human T-cell lines with well-defined T-cell receptor gene rearrangements as controls for the BIOMED-2 multiplex polymerase chain reaction tubes.
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.
authors.affiliation: Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.
authors.forename: Y
authors.initials: Y
authors.lastname: Sandberg
authors.forename: B
authors.initials: B
authors.lastname: Verhaaf
authors.forename: E J
authors.initials: EJ
authors.lastname: van Gastel-Mol
authors.forename: I L M
authors.initials: IL
authors.lastname: Wolvers-Tettero
authors.forename: J
authors.initials: J
authors.lastname: de Vos
authors.forename: R A F
authors.initials: RA
authors.lastname: Macleod
authors.forename: J G
authors.initials: JG
authors.lastname: Noordzij
authors.forename: W A
authors.initials: WA
authors.lastname: Dik
authors.forename: J J M
authors.initials: JJ
authors.lastname: van Dongen
authors.forename: A W
authors.initials: AW
authors.lastname: Langerak
mesh_terms: D018929:Cell Culture Techniques; D002460:Cell Line; D015321:Gene Rearrangement; D006801:Humans; D016130:Immunophenotyping; D016133:Polymerase Chain Reaction / Q000379:methods*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D011948:Receptors, Antigen, T-Cell