title: Evaluation of T-cell clonality in archival skin biopsy samples of cutaneous T-cell lymphomas using the biomed-2 PCR protocol.
abstract: Recently, several European centers of lymphoma diagnosis and research developed various polymerase chain reaction (PCR) methods for clonality analysis in suspect T-cell and B-cell proliferations (Biomed-2 Concerted Action). They have mainly been applied to frozen material of systemic B-cell and T-cell malignancies. Thus far, only limited data exist with regard to cutaneous T-cell lymphoma (CTCL) and paraffin-embedded material. Thus, we applied the Biomed-2 T-cell receptor (TCR) gamma and TCRbeta PCR as well as an in-house TCRgamma PCR to a collection of 107 archival skin samples (84 CTCL, 3 systemic TCL and 20 controls). As a result, the Biomed-2 TCRgamma PCR revealed 81% clonality, the in-house TCRgamma method revealed 86% clonality, and the Biomed-2 TCRbeta revealed 78% clonality in CTCL samples generating at least the 300 bp fragment in the Biomed-2 control PCR. We found clonal TCRbeta rearrangements in 5 of 17 CTCL samples that were polyclonal in the Biomed-2 TCRgamma PCR. By combining all Biomed-2 assays, one or more clonal rearrangements were detected in 87% of CTCL and in all 3 systemic TCLs. By combining all TCR PCR assays applied here, clonality was shown in 90% of the CTCL cases. In conclusion, we showed that the Biomed-2 TCR PCR worked well with DNA from paraffin-embedded tissue, revealing a high-clonality detection rate in CTCL, and thus should be highly recommended for routine molecular analysis. In addition, the performance of our in-house TCRgamma assay verifies our previously published findings on clonally expanded T-cells in CTCL.
authors.affiliation: Department of Dermatology and Allergy, Campus Benjamin Franklin, Charité-Universitätsmediz, Berlin, Germany. ansgar.lukowsky@charite.de
authors.forename: Ansgar
authors.initials: A
authors.lastname: Lukowsky
authors.forename: J Marcus
authors.initials: JM
authors.lastname: Muche
authors.forename: Markus
authors.initials: M
authors.lastname: Möbs
authors.forename: Chalid
authors.initials: C
authors.lastname: Assaf
authors.forename: Daniel
authors.initials: D
authors.lastname: Humme
authors.forename: Michael
authors.initials: M
authors.lastname: Hummel
authors.forename: Wolfram
authors.initials: W
authors.lastname: Sterry
authors.forename: Matthias
authors.initials: M
authors.lastname: Steinhoff
mesh_terms: D001706:Biopsy; D006801:Humans; D016410:Lymphoma, T-Cell, Cutaneous / Q000175:diagnosis* / Q000473:pathology; D016612:Paraffin Embedding; D057089:Pathology, Molecular / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics; D012680:Sensitivity and Specificity; D012867:Skin / Q000473:pathology*; D013601:T-Lymphocytes
chemical_list: D016693:Receptors, Antigen, T-Cell, alpha-beta; D016692:Receptors, Antigen, T-Cell, gamma-delta