title: Separation and analysis of mononuclear cells infiltrating the thyroid of patients with Graves' disease.
abstract: A simple method was established for separating lymphocytes infiltrating the thyroid from thyroid epithelial cells. Namely, suspensions of minced thyroid from patients with Graves' disease were layered on a Percoll two-step density gradient (p = 1.050 and 1.077 g/ml) and centrifuged (400g, 30 min, 4 degrees C). In this way 0.1-18 X 10(5) lymphocytes/g of thyroid tissue with a purity of 65-95% were obtained. Thyroid lymphocytes were analyzed quantitatively with monoclonal antibodies by laser flow cytometry and compared with peripheral lymphocytes. The proportion of OKT3+ cells was decreased with increase in OKIa+ cells. The percentage of OKIa+ cells was significantly correlated with that of Leu12+ cells. The percentages of OKT4+ cells and OKIa+ cells were higher when analyzed with an extended gate window, which was arranged for detection of activated, large-sized lymphocytes. The percentages of OKT8+ and Leu7+ cells were not significantly different from those in peripheral blood. From these results it was concluded that the proportion of B lymphocytes is increased and that of T lymphocytes is decreased, the proportion of activated B lymphocytes is increased, some helper/inducer T cells are activated in the thyroid gland in Graves' disease, and these activated lymphocytes may be important in local production of antithyroid autoantibodies.
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal; D002469:Cell Separation / Q000379:methods; D004847:Epithelial Cells; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006111:Graves Disease / Q000097:blood* / Q000473:pathology; D006801:Humans; D007834:Lasers; D008214:Lymphocytes / Q000166:cytology*; D008297:Male; D013961:Thyroid Gland / Q000473:pathology*
chemical_list: D000911:Antibodies, Monoclonal