A POLYCLONAL T-CELL REPERTOIRE OF V-ALPHA AND V-BETA T-CELL RECEPTOR GENE FAMILIES IN INTRATHYROIDAL T-LYMPHOCYTES OF GRAVES-DISEASE PATIENTS

We have examined for the presence or absence of T cell receptor V-alph a (VA) and V-beta (VB) gene expression in infiltrating T lymphocytes ( ITL) isolated from Graves' thyroid glands in comparison to paired peri pheral blood lymphocyte (PBL) samples using a qualitative based polyme rase chain reaction (PCR) assay. Sequence specific oligonucleotides fo r VA and VB T cell receptor gene (TCR) families that had previously be en validated in other studies, were used for the PCR analysis, followe d by Southern blot hybridization with a labelled, internal C-region pr imer. A total of seven Graves' disease patients who had been treated w ith carbimazole were studied. T cell receptor VA and VB gene usage was examined in freshly isolated, unstimulated ITLs from five patients. A widespread usage of VA and VB gene families with 12 to 18 families be ing used was apparent. Use of oligo-dT or C-region priming of the mRNA prior to reverse transcription of the mRNA did not have any significa nt affect on the results nor did the use of whole Graves' thyroid mRNA as the starting material (n = 2) or perfusion of one gland with salin e to remove as much of the contaminating blood from the gland. Our res ults contrast with those of Davies and colleagues who have previously shown a restricted repertoire of VA gene families in ITLs in compariso n to autologous PBLs, and are much more in line with other recent repo rts indicating a diverse VA repertoire of the infiltrating T cells in Graves' thyroid glands derived from patients treated with anti-thyroid drugs.