A strategy for demonstrating the clonal origin of small numbers of T lymphocytes in histopathological specimens

J Pathol. 1989 Jul;158(3):261-5. doi: 10.1002/path.1711580315.

Abstract

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.

MeSH terms

  • Base Sequence
  • Clone Cells*
  • DNA-Directed DNA Polymerase / metabolism
  • Gene Amplification*
  • Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor
  • Humans
  • Molecular Sequence Data
  • T-Lymphocytes / pathology*

Substances

  • DNA-Directed DNA Polymerase