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中文摘要：
目的观察高迁移率族蛋白B1（HMGB1）对调节性T细胞（Treg）与CD4^＋CD25^-T细胞相互作用的影响，并初步探讨其影响Treg抑制功能的机制。方法免疫磁珠法分离正常BALB／c小鼠脾脏CD4^＋CD25^-Treg及CD4^＋CD25^-T细胞。采用固相包被抗CD3／可溶性抗CD28进行辅助活化，以不同时间及浓度HMGB1刺激Treg，ELISA法分析HMGB1刺激对Treg分泌IL-10的影响。将HMGB1（1000μg／L）刺激后的Treg与CD4^＋CD25^-T细胞共培养，MTT法观察其对Treg抑制CD4^＋CD25^-T细胞反应的作用，并分析HMGB1刺激后的Treg对CD4^＋CD25^-T细胞IL-2生成及细胞功能极化的影响。结果经抗CD3／CD28辅助活化的CD4^＋CD25^-Treg在HMGB1作用下IL-2生成无显著差异（P〉0．05），但随时间延长及剂量增加，IL-10生成明显减少（P〈0．05）。经HMGB1刺激的Treg对CD4^＋CD25^-T细胞增殖的抑制反应减弱，同时诱导CD4^＋CD25^-T细胞IL-2产生及细胞功能极化的能力均下降（P〈0．05）。结论HMGB1可通过诱导Treg抑制功能的下调，从而影响CD4^＋CD25^-T细胞功能，进而调节炎症反应过程。
英文摘要：
Objective To investigate the influence of high mobility group box-1 protein （ HMGB1 ） on the immunosuppression function of splenic regulatory T cells （Tregs） and its potential regulatory mechanism underlying the effect on CD4^＋ CD25 ^- T cells in mice. Methods CD4^＋ CD25^- Tregs isolated from the spleens of male BALB/c mice by magnetic beads were seeded on 96-well （ 1 × 10^5 cells/well） cell culture plates coated with 1 μg/ml anti-CD3 and soluble CD28. After being stimulated with HMGB1 for different time and concentrations, the secretions of IL-2 and IL-10 were analyzed by ELISA. Tregs stimulated for 72 hours were cultured with CD4^＋ CD25 - T cells together. The suppressive activity of CD4^＋ CD25^- Treg to CD4^＋ CD25 ^- T cells was analyzed by MTT test. IL-2, IL-10, IL-4, and interferon （IFN）-γin the cell suspensions were determined by ELISA. Results After stimulation with HMGB1, the suppressive activity of splenic Tregs in mice were significantly down-regulated at 72 hours, when the proportion of Tregs to CD4^＋ CD25 ^- T cells was 1 ： 1. The secretion of IL-2 of Tregs stimulated by HMGB1 was not markedly changed （ P 〉 0. 05 ）, while a dose-dependent decrease between IL-10 induction and HMGB1 concentration was obviously （ P 〈 0.05 ）. When CD4^＋ CD25^- T cells were cultured with stimulated Tregs, comparing with unstimulated-Treg group, levels of IL-2 and IFN-γ were elevated following the increased concentration of HMGB1 （P 〈 0.05 or P 〈 0. 01 ）. Meanwhile the secretion of IL-4 and IL-10 significantly decreased when cultured with stimulated Tregs （ P 〈 0. 05 ）. Condusions These data suggested that HMGB1 stimulation can result in significant down-regulation of immunosuppression of splenic Tregs in mice. HMGB1 might be a potential immunoregnlatory signal that influences the proliferation of effector T cells, secretion of IL-2 and cells-polarization by inhibiting CD4^＋ CD25^- Tregs activity.
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