Objective: To detect the rearrangement of T cell receptor gamma chain gene in single H/R-S cell as an indication of early T lymphocyte differentiation.
Methods: CD30 positive H/R-S cells were isolated with micromanipulator from the frozen sections and analyzed by single-cell PCR. TCR-gamma chain gene rearrangement was detected through semi-nested PCR. In the first round of PCR, a pair of TCR-gamma primers which amplify the segments between Vgamma and Jgamma region (including the N region) of TCR-gamma gene mixing with a pair of beta-globin primers were used. In the second round, only the products positive beta-globin were amplified with the primers for TCR-gamma gene.
Results: A 268 bp beta-globin gene specific product was obtained in 24 out of 56 H/R-S cells from 6 cases of Hodgkin's disease. After the first round of PCR, no rearrangement of the TCR-gamma gene was noticed at a single cell level.
Conclusion: H/R-S cells from the selected cases were not derived from the T-cell lineage but possibly from the B-cell lineage.
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