Polymerase chain reaction amplification analyses of clonality in T-cell malignancy including peripheral T-cell lymphoma.
Clonality in T-cell malignancy was investigated using T-cell receptor (TcR) V beta 1-20 family primers and polymerase chain reaction amplification (PCR) of cDNA prepared from tissue biopsies and blood involved with tumour. Secondary PCR amplification of the VDJ joints of primary PCR products was performed to distinguish clonal from polyclonal products, and clonal V beta gene products were confirmed by direct PCR sequencing in the majority of cases. In eight T-cell malignancies including T-cell acute lymphoblastic leukaemia (T-ALL) and T-cell chronic lymphocytic leukaemia (T-CLL) shown to be clonal by Southern blot analysis, one or two primary PCR products were identified and shown to be clonal. In five cases of peripheral T-cell lymphoma (PTCL) all V beta 1-20 families were identified after primary PCR amplification, and clonal products were identified in two cases after secondary amplification; TcR V beta clonal families could not be demonstrated in the remaining three cases. These data were in agreement with previous Southern blot analysis of these cases, and confirmed the presence of reactive T cells in PTCL as well as providing further evidence for the genotypic heterogeneity of this entity. In the remaining case, a blood lymphocytosis, primary PCR amplification produced predominant TcR V beta 6 and V beta 12 family products, of which the V beta 6 family proved clonal after secondary PCR amplification. There was no evidence for overrepresentation of TCR V beta families by the tumour populations in this study, furthermore the data confirm the involvement of reactive cells in T-cell malignancy and the genetic heterogeneity of PTCL.
Hodges
E
E
Molecular Immunology Group, Southampton University Hospitals NHS Trust, UK.
Edwards
S E
SE
Howell
W M
WM
Smith
J L
JL
D001483:Base Sequence; D015139:Blotting, Southern; D015342:DNA Probes; D005784:Gene Amplification; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D005838:Genotype; D006801:Humans; D015458:Leukemia, T-Cell / Q000235:genetics*; D016411:Lymphoma, T-Cell, Peripheral / Q000235:genetics*; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D015342:DNA Probes