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首页> 外文期刊>Autoimmunity >Application of mouse monoclonal antibodies for identification of antigen regions of human thyroid peroxidase in adolescents with Graves' disease and non-toxic multinodular goiter by flow cytometry.
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Application of mouse monoclonal antibodies for identification of antigen regions of human thyroid peroxidase in adolescents with Graves' disease and non-toxic multinodular goiter by flow cytometry.

机译：小鼠单克隆抗体在流式细胞仪中鉴定患有Graves病和无毒多结节性甲状腺肿的青少年中人甲状腺过氧化物酶抗原区域的应用。

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Thyroid peroxidase (TPO) is the major thyroid autoantigen recognized by serum autoantibodies from patients with Graves' disease (GD) or Hashimoto's thyroiditis directed to two immunodominant conformational regions termed A and B. The epitopes of human TPO have been defined using a panel of mouse monoclonal antibodies (mAbs). The aim of this study was to estimate the expression of chosen surface antigen regions of TPO (1, 18, 30, 64 epitopes) on thyroid cells in 15 patients with non-toxic multinodular goiter (NTMG) and 15 patients with GD. The thyrocytes were identified by indirect method: in the first stage we added mouse monoclonal autoantibodies specific for TPO regions and in the second stage we conjugated this complex with rabbit anti-mouse antibodies IgG (Fab')(2) with FITC. All investigations were performed by flow cytometry using Coulter EPICS XL apparatus. The percentages of thyrocytes with expression of epitopes 1, 18, 30, 64 TPO were measured in relation to the respective anti-TPO concentrations: 50-1600 microg/ml. The analysis of epitopes located in immunodominant regions (IDR) of TPO revealed higher percentages of thyrocytes in cases with GD in comparison to NTNG. The most predominant difference was observed for mAb 64 epitope (48 vs 7%, p < 0.019; 39 vs 5%, p < 0.017) at the concentration of 100-200 microg/ml mAbs. The expression of 18 epitope on thyrocytes was also statistically higher in Graves' patients than in the NTMG (14 vs 6%, p < 0.025) at concentration of 400 microg/ml mAbs. However, this expression was much less pronounced. In all the cases, the percentages of thyrocytes with epitopes 1 and 30 were in low detection (8-15% of positive cells). In conclusion, our findings suggest that the elevated expression of TPO epitopes 18 and 64 in young patients with thyroid autoimmune diseases increase stimulation and activation of thyroid cells during inflammatory reaction within the thyroid gland. In addition, predominant expression of 64 TPO epitope that recognizes B domain in GD patients could be a useful marker of the immune process in the thyroid gland.
机译：甲状腺过氧化物酶（TPO）是被Graves病（GD）或Hashimoto甲状腺炎患者的血清自身抗体识别的主要甲状腺自身抗原，这些抗原针对被称为A和B的两个免疫优势构象区。已使用一组小鼠定义了人类TPO的表位单克隆抗体（mAbs）。这项研究的目的是评估15例无毒多结节性甲状腺肿（NTMG）和15例GD患者的甲状腺细胞中TPO选定表面抗原区域（1、18、30、64个表位）的表达。甲状腺细胞通过间接方法鉴定：在第一阶段，我们添加了对TPO区具有特异性的小鼠单克隆自身抗体，在第二阶段，我们将该复合物与兔抗小鼠抗体IgG（Fab'）（2）与FITC偶联。所有研究均使用Coulter EPICS XL仪器通过流式细胞仪进行。相对于各自的抗-TPO浓度：50-1600μg/ ml，测量了具有表位1、18、30、64 TPO表达的甲状腺细胞的百分比。对TPO免疫优势区（IDR）中抗原决定簇的分析表明，与NTNG相比，GD患者的甲状腺细胞百分比更高。在100-200 microg / ml mAb的浓度下，观察到mAb 64表位的最主要差异（48对7％，p <0.019; 39对5％，p <0.017）。在浓度为400微克/毫升mAbs的情况下，格雷夫斯病患者的甲状腺细胞中18个表位的表达在统计学上也高于NTMG（14 vs 6％，p <0.025）。但是，这种表达不太明显。在所有情况下，具有表位1和30的甲状腺细胞百分比均为低检测值（占阳性细胞的8-15％）。总之，我们的研究结果表明，在甲状腺自身免疫性疾病的年轻患者中，TPO表位18和64的表达升高，在甲状腺内炎症反应过程中增加了对甲状腺细胞的刺激和激活。此外，GD患者中识别B结构域的64个TPO表位的主要表达可能是甲状腺免疫过程的有用标志。

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