A strategy for demonstrating the clonal origin of small numbers of T lymphocytes in histopathological specimens.
A strategy is described for detecting large T-cell clones among small numbers of lymphoid cells in fresh or formalin-fixed tissue samples using the polymerase chain reaction (PCR) to amplify and identify rearrangements of the V and J genes of the T-cell gamma receptor. Following hybridization of primers with gene-specific sequences at judiciously selected locations on each of the eight potentially active V and five potentially active J genes, the PCR can theoretically amplify DNA segments which span the join between rearranged V and J genes and are of approximately 384 different sizes, each segment size reflecting different gamma gene clonal rearrangements. Large monoclonal populations of T lymphocytes, indicated by excessive amounts of particular PCR segment sizes, can be further characterized by direct nucleotide sequencing of the hypervariable N regions of these segments, and the presence of such clones can be confirmed directly in tissue sections by in situ hybridization with N region-specific oligonucleotide probes.
Goudie
R B
RB
University Department of Pathology, Glasgow Royal Infirmary, U.K.
D001483:Base Sequence; D002999:Clone Cells; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D005784:Gene Amplification; D015334:Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; D006801:Humans; D008969:Molecular Sequence Data; D013601:T-Lymphocytes / Q000473:pathology*
D004259:DNA-Directed DNA Polymerase