Ueki Y, Eguchi K, Fukuda T, Otsubo T, Kawabe Y, Shimomura C, Matsunaga M, Tezuka H, Ishikawa N, Ito K
First Department of Internal Medicine, Nagasaki University School of Medicine, Japan.
J Clin Endocrinol Metab. 1987 Nov;65(5):922-8. doi: 10.1210/jcem-65-5-922.
To investigate the suppressor function of intrathyroidal (TG) T cells in Graves' disease, the percentage of suppressor T cell subsets and the suppressor function of TG and peripheral blood (PB) lymphocytes in Graves' disease were compared by determining the in vitro production of immunoglobulin G (IgG) in reconstituted mixtures of separated B, CD4+ (helper/suppressor-inducer T), and CD8+ (suppressor/cytotoxic T) cells. TG lymphocytes were obtained by gradient centrifugation of the supernatants of minced thyroid tissues. T Cells were separated by E-rosette formation, and CD4+ and CD8+ cell-rich populations were separated by a panning method using monoclonal anti-CD8 antibody. Mixtures of 5 X 10(4) B (PB1 or TG) cells, 2 X 10(4) CD4+ (PB or TG) cells, and 5 X 10(3) macrophages (PB or TG) were cultured with various numbers of CD8+ (PB) or CD8+ (TG) cells for 7 days with pokeweed mitogen, and IgG synthesis was determined by solid phase RIA. T cell subpopulations were quantitated by a direct immunofluorescence method using monoclonal anti-CD3, anti-CD4, and anti-CD8 antibodies. There was no difference in the percentages of CD8+ cells among total T cells between thyroid glands and peripheral blood from Graves' disease patients [mean PB, 39.8 +/- 9.8% (+/- SD); TG, 42.5 +/- 13.8%; n = 10]. IgG production by mixtures of B and CD4+ cells isolated from peripheral blood was not different from that by cells isolated from thyroid glands [mean PB, 1635 +/- 248 (+/- SE) ng/mL; TG, 1081 +/- 128 ng/mL; n = 19; P = NS]. The nonspecific suppressor activity of thyroid gland CD8+ cells was less than that of CD8+ (PB) cells [percent suppression of IgG production by mixtures of B (PB) and CD4+ (PB) cells, 12.5% vs. 57.0% (P less than 0.01); by mixtures of B (TG) and CD4+ (TG) cells, 5.8% vs. 38.9% (P less than 0.01)]. The suppressor-inducer function of CD4+ (TG) cells was also decreased compared with that of CD4+ (PB) cells. These results suggest that the impairment of suppressor cell activity may lead to excessive production of autoantibody in thyroid glands from patients with Graves' disease.
为研究格雷夫斯病中甲状腺内（TG）T细胞的抑制功能，通过测定分离的B细胞、CD4 +（辅助/抑制诱导性T）细胞和CD8 +（抑制/细胞毒性T）细胞重组混合物中免疫球蛋白G（IgG）的体外产生情况，比较了格雷夫斯病中TG和外周血（PB）淋巴细胞的抑制性T细胞亚群百分比及抑制功能。TG淋巴细胞通过对切碎的甲状腺组织上清液进行梯度离心获得。T细胞通过E花环形成进行分离，富含CD4 +和CD8 +细胞的群体通过使用单克隆抗CD8抗体的淘选法进行分离。将5×10⁴个B（PB1或TG）细胞、2×10⁴个CD4 +（PB或TG）细胞和5×10³个巨噬细胞（PB或TG）的混合物与不同数量的CD8 +（PB）或CD8 +（TG）细胞一起用商陆有丝分裂原培养7天，并用固相放射免疫分析法测定IgG合成。使用单克隆抗CD3、抗CD4和抗CD8抗体通过直接免疫荧光法对T细胞亚群进行定量。格雷夫斯病患者甲状腺和外周血中总T细胞中CD8 +细胞的百分比没有差异[平均PB，39.8±9.8%（±标准差）；TG，42.5±13.8%；n = 10]。从外周血分离的B细胞和CD4 +细胞混合物产生的IgG与从甲状腺分离的细胞产生的IgG没有差异[平均PB，1635±248（±标准误）ng/mL；TG，1081±128 ng/mL；n = 19；P =无显著性差异]。甲状腺CD8 +细胞的非特异性抑制活性低于CD8 +（PB）细胞[B（PB）和CD4 +（PB）细胞混合物对IgG产生的抑制百分比，12.5%对57.0%（P＜0.01）；B（TG）和CD +（TG）细胞混合物，5.8%对38.9%（P＜0.01）]。与CD4 +（PB）细胞相比，CD4 +（TG）细胞的抑制诱导功能也降低。这些结果表明，抑制细胞活性受损可能导致格雷夫斯病患者甲状腺中自身抗体过度产生。
