Bossowski Artur, Stasiak-Barmuta Anna, Urban Mirosława
2nd Department of Children's Diseases, Medical University of Białystok, Białystok, Poland.
Horm Res. 2005;64(4):189-97. doi: 10.1159/000088875. Epub 2005 Oct 10.
The present study was performed to elucidate the relationship between CTLA-4/CD28 molecules and stimulating (TSAb) and blocking (TBAb) antibodies to the TSH-receptor (TSH-R) in Graves' disease. CD28 and CD152 (CTLA-4) are glycoprotein molecules which provide a potent costimulatory signal for T-cell activation and proliferation via interactions with their ligands, B7.1/B7.2 molecules, which are present on the surface of antigen-presenting cells. The aim of the study was to estimate the expression of cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4, CD152), CD28, B7.1 (CD80), and B7.2 (CD86) molecules on peripheral blood cells in patients with Graves' disease (GD) (n = 55, mean age 15.5 +/- 5.1 years) and nontoxic nodular goiter (NTNG) (n = 55, mean age 15.2 +/- 4.5 years), in comparison with sex and age-matched healthy control subjects (n = 55, mean age 15.2 +/- 3.9 years). The expression of the costimulatory molecules on mononuclear cells was analyzed by three-color flow cytometry using a Coulter EPICS XL cytometer. Detection of TSAb and TBAb to the TSH-R using JPO9 CHO cells in unfractionated serum was measured by a highly sensitive commercial radioimmunoassay. When compared with healthy control subjects and euthyroid patients with GD, untreated patients with GD showed a significant increase of CD152+ (p < 0.001, p < 0.001) and CD28+ (p < 0.01, NS) T lymphocytes, respectively. After 6-12 months of methimazole therapy, the percentage of these cells in the peripheral blood of hyperthyroid patients returned to normal values. In addition, patients with GD showed an increase in the percentage of both B7.1 (3.8%) and B7.2 (18.4%) molecules on activated monocytes, compared to patients with NTNG (0.5% p < 0.05, 2.5% p < 0.01, respectively) and healthy control subjects (0.2% p < 0.05, 0.8% p < 0.003, respectively). In patients with untreated GD there was a statistically significant positive correlation between the expression of CTLA-4 on the surface of peripheral blood T cells and the index of TSAb antibodies (R = 0.54, p < 0.001) as well as a negative correlation with TBAb antibody titer (R = -0.58, p < 0.001). However, no such correlations were noted with regard to CD28 and anti-TPO, anti-TG, and TRAb antibodies. We conclude that changes in the expression of costimulatory molecules on the surface of peripheral blood T cells and their significant relationship with the level of antithyroid antibodies indicate an involvement of these molecules in the pathogenesis of GD.
本研究旨在阐明格雷夫斯病中CTLA-4/CD28分子与促甲状腺素受体（TSH-R）刺激抗体（TSAb）和阻断抗体（TBAb）之间的关系。CD28和CD152（CTLA-4）是糖蛋白分子，它们通过与抗原呈递细胞表面存在的配体B7.1/B7.2分子相互作用，为T细胞活化和增殖提供强大的共刺激信号。本研究的目的是评估格雷夫斯病（GD）患者（n = 55，平均年龄15.5 +/- 5.1岁）和非毒性结节性甲状腺肿（NTNG）患者（n = 55，平均年龄15.2 +/- 4.5岁）外周血细胞上细胞毒性T淋巴细胞相关抗原-4（CTLA-4，CD152）、CD28、B7.1（CD80）和B7.2（CD86）分子的表达，并与性别和年龄匹配的健康对照受试者（n = 55，平均年龄15.2 +/- 3.9岁）进行比较。使用库尔特EPICS XL细胞仪通过三色流式细胞术分析单核细胞上共刺激分子的表达。通过高灵敏度商业放射免疫测定法，使用JPO9 CHO细胞在未分离的血清中检测TSH-R的TSAb和TBAb。与健康对照受试者和GD甲状腺功能正常的患者相比，未经治疗的GD患者的CD152+（p < 0.001，p < 0.001）和CD28+（p < 0.01，无显著性差异）T淋巴细胞分别显著增加。甲巯咪唑治疗6 - 12个月后，甲状腺功能亢进患者外周血中这些细胞的百分比恢复到正常水平。此外，与NTNG患者（分别为0.5% p < 0.05，2.5% p < 0.01）和健康对照受试者（分别为0.2% p < 0.05，0.8% p < 0.003）相比，GD患者活化单核细胞上B7.1（3.8%）和B7.2（18.4%）分子的百分比均增加。在未经治疗的GD患者中，外周血T细胞表面CTLA-4的表达与TSAb抗体指数之间存在统计学显著正相关（R = 0.54，p < 0.001），与TBAb抗体滴度呈负相关（R = -0.58，p < 0.001）。然而，未发现CD28与抗TPO、抗TG和TRAb抗体之间存在此类相关性。我们得出结论，外周血T细胞表面共刺激分子表达的变化及其与抗甲状腺抗体水平的显著关系表明这些分子参与了GD的发病机制。
