Molecular biological approach to allergy disease medical treatment

目的：本研究的目的是探讨CD25^<高> CD4+Tregs的功能，包括调节CD25^-T细胞活化和Ca^<2+>对T细胞受体（TCR）刺激的反应，作为一种能量标志，在哮喘中是否发生改变。方法：采集昭和大学藤冈医院和昭和大学门诊患者的血液样本。用荧光显像技术分析Ca^<2+>的响应。流式细胞术检测细胞增殖情况。结果：非哮喘患者CD25^<高> CD4+ treg对TCR刺激无反应，导致细胞增殖不足和Ca^<2+>反应。然而，与非哮喘患者相比，哮喘患者的CD25^<高> CD4+ treg对刺激有反应，且Ca^<2+>的反应模式可分为两组，1型和2型。1型细胞大多数细胞表现出正常的Ca^<2+>反应，并且与正常细胞相比，表现出强增殖和较弱的调节功能。相比之下，2型细胞反应性较好，但细胞内Ca^<2+>略有增加，并部分发挥调节细胞的功能。结论：我们发现非哮喘患者CD25^<高>的CD4+ treg细胞中的Ca^<2+>反应受损，而哮喘患者细胞中的Ca^<2+>反应在TCR作用下增加。这些观察结果表明，哮喘患者的CD25^<高>CD4+ Tregs异常与气道慢性炎症有关。

Objective : The aim of this study was to investigate whether the functions of the CD25^<high> CD4+Tregs including regulation of CD25^-T cells activation and the Ca^<2+> response to T cell receptor (TCR) stimulation as a marker of anergy are changed in asthma. Methods: Blood samples were collected from the outpatients of Showa University Fujigaoka Hospital and Showa University. Ca^<2+> response were analyzed with fluoroimaging technique. Proliferation was analyzed by flow cytometry.Results : The CD25^<high> CD4+ Tregs from non-asthmatics were unresponsive to TCR stimulation, resulting in paucity of proliferation and Ca^<2+> response. However, in contrast to non-asthmatics, the CD25^<high> CD4+ Tregs from asthmatics were responsive to the stimulation, and the Ca^<2+> response pattern could be classified into two groups, Type 1 and Type 2. Type 1 cells exhibited normal Ca^<2+> response in most of cells, and showed enforced proliferation and weaker regulatory functions compared to the normal ones. In contrast, Type 2 cells were responsive but had a slight increase in intra-cellular Ca^<2+>, and partially functioned as regulatory cells.Conclusion : We identified impaired Ca^<2+> responses in CD25^<high>CD4+ Tregs from non-asthmatics but increasing Ca^<2+> responses in the cells from asthmatics upon TCR engagement. These observations suggested that abnormalities of CD25^<high>CD4+ Tregs in asthmatics are implicated in the chronic inflammation in the airway.