Akasu F, Kasuga Y, Matsubayashi S, Carayon P, Volpé R
Endocrinology Research Laboratory, Wellesley Hospital, University of Toronto, Ontario, Canada.
Thyroid. 1991 Summer;1(3):215-22. doi: 10.1089/thy.1991.1.215.
We have studied by flow cytometric analysis the antigen specific activation of CD4+ (helper/inducer) T lymphocytes by purified human thyroid peroxidase (TPO). Peripheral blood mononuclear cells were obtained from 26 patients with Graves' disease (GD), 16 with Hashimoto's thyroiditis (HT), 7 with nontoxic nodular goiter (NG), and 14 normal subjects (N). Cells were cultured for 7 days in the presence or absence of TPO at final concentrations of 3, 30, and 300 ng/mL. When harvested, cells were reacted with an FITC-conjugated anti-CD4 and a PE-conjugated anti-HLA-DR murine monoclonal antibodies. The percentage of HLA-DR+ CD4+ cells (activated CD4+ cells) was determined by a flow cytometer. In the absence of TPO, CD4+ cells had been activated without any specific stimulant. This is known as the autologous mixed lymphocyte reaction (AMLR). In the AMLR, CD4+ cells from GD and HT were less activated compared to those from NG and N. Results of TPO-specific activation were expressed as an incremental increase of activated CD4+ cells (II) (percentage of activated CD4+ cells cultured with TPO minus percentage of activated CD4+ cells cultured without TPO). II of N, GD, HT, and NG were 0.37 +/- 0.21, 2.20 +/- 0.45,** 2.0 +/- 0.66,* and 0.35 +/- 0.27 (mean +/- SEM), respectively (**p less than 0.01; *p less than 0.05 vs N). When patients were further subdivided, the highest mean II was found in patients with hyperthyroid GD (p less than 0.01), followed by euthyroid HT (p less than 0.05) and euthyroid GD (p less than 0.05), however there was no significant difference between hypothyroid HT and N. In conclusion (1) AMLR reactivity of CD4+ cells from GD and HT was impaired, (2) however, CD4+ cells from both GD and HT were significantly more induced by TPO compared to N, and (3) this induction depends, in part, on the in vivo thyroid status.
我们通过流式细胞术分析研究了纯化的人甲状腺过氧化物酶（TPO）对CD4 +（辅助/诱导）T淋巴细胞的抗原特异性激活作用。从26例格雷夫斯病（GD）患者、16例桥本甲状腺炎（HT）患者、7例非毒性结节性甲状腺肿（NG）患者和14名正常受试者（N）获取外周血单个核细胞。细胞在终浓度为3、30和300 ng/mL的TPO存在或不存在的情况下培养7天。收获细胞时，使其与异硫氰酸荧光素（FITC）偶联的抗CD4和藻红蛋白（PE）偶联的抗HLA-DR鼠单克隆抗体反应。通过流式细胞仪测定HLA-DR + CD4 +细胞（活化的CD4 +细胞）的百分比。在没有TPO的情况下，CD4 +细胞在没有任何特异性刺激物的情况下被激活。这被称为自体混合淋巴细胞反应（AMLR）。在AMLR中，与来自NG和N的细胞相比，来自GD和HT的CD4 +细胞活化程度较低。TPO特异性激活的结果表示为活化的CD4 +细胞的增量增加（II）（用TPO培养的活化的CD4 +细胞百分比减去不用TPO培养的活化的CD4 +细胞百分比）。N、GD、HT和NG的II分别为0.37±0.21、2.20±0.45**、2.0±0.66*和0.35±0.27（平均值±标准误）（**p<0.01；*p<0.05，与N相比）。当患者进一步细分时，甲状腺功能亢进的GD患者的平均II最高（p<0.01），其次是甲状腺功能正常的HT患者（p<0.05）和甲状腺功能正常的GD患者（p<0.05），然而甲状腺功能减退的HT患者与N之间没有显著差异。总之，（1）来自GD和HT的CD4 +细胞的AMLR反应性受损，（2）然而，与N相比，来自GD和HT的CD4 +细胞被TPO诱导的程度明显更高，并且（3）这种诱导部分取决于体内甲状腺状态。
