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The peripheral activated T-cell subsets which express concurrently DR and CD antigens were determined in 22 patients with Graves' disease using the fluorescence-rosette double labelling technique. In the active phase of the disease a decrease in the percentage of CD8+ cells, an increase in the CD4+/CD8+ ratio and an elevation of various DR+CD+T-lymphocytes was found. A concurrent marked increase in DR+CD4+ cells accounts for an abnormality in the equilibrium of activated T-lymphocyte subsets and an increase in DR+CD4+/DR+CD8+ ratios. In comparison with the active phase the DR+CD3+ and DR+CD4+ cells decreased in remission, but DR+CD8+ cells remained unchanged. The decrease in the ratio of CD4+/CD8+ cells was less marked than that of the DR+CD4+/DR+CD8+ cells, which fell into the normal range. Our results suggest an excessive activation of T-helper cells as well as a deficiency in T-suppressor cells which may play an important role in the pathogenesis of Graves' disease.

In order to determine the concurrent expression of DR and CD antigen of T lymphocyte surface markers, the fluorescence-rosette double-labelling technique was developed by employing monoclonal antibody against HLA-DR and sensitized sheep erythrocytes with OKT series of monoclonal antibodies against cell surface antigen. Its reliability was evaluated by comparison with two-step isolating technique for determining DR antigen of activated T lymphocyte subsets in 30 healthy volunteers. The result showed there was no statistic difference between results determined with two methods. The double-labelling technique was characterized by its samplity, no injury for cellular activity and ability for analysing the expression of the single antigen and double antigens on cell surface concurrently.