Cheng Xiang, Chen Yong, Xie Jiang-Jiao, Yao Rui, Yu Xian, Liao Meng-Yang, Ding Ying-Jun, Tang Ting-Ting, Liao Yu-Hua, Cheng Yan
Laboratory of Cardiovascular Immunology, Institute of Cardiology, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430022, China.
J Mol Cell Cardiol. 2008 Aug;45(2):168-75. doi: 10.1016/j.yjmcc.2008.04.003. Epub 2008 Apr 18.
Atherosclerosis is a chronic inflammatory disease, which is positively and negatively regulated by T helper (Th) 1 and Th2 lymphocytes, respectively. Recent findings indicate that suppressive oligodeoxynucleotides (ODNs) expressing TTAGGG motifs selectively reduce Th1 cytokine production and have been proven effective at blocking the development of organ-specific autoimmune diseases. In the current research, we hypothesized that suppressive ODNs may alter the development of atherosclerosis. Eight-week-old homozygous ApoE(-/-) male mice were injected with 300 mug ODNs A151 (TTAGGG) or nonspecific ODNs 1612. Atherosclerotic lesion sizes were dramatically reduced by ODNs A151, but not by nonspecific ODNs. MCP-1 and VCAM-1, which are the key inflammatory factors in atherogenesis, were significantly attenuated by the suppressive ODNs A151. In the splenic lymphocytes, FACS analysis showed ODNs A151 reduced the percentage of IFN-gamma-producing Th1 cells and slightly increased the percentage of IL-4-producing Th2 cells, indicating that suppressive ODNs skewed the Th1/Th2 balance toward Th2 inflammation in vivo. Furthermore, ODNs A151 down-regulated the phosphorylation of STAT1 and STAT4 and suppressed up-regulation of T-bet, a signal modulator for Th1, and didn't impact GATA-3 and STAT6, which are associated with a Th2 phenotype. Consistent with this in vivo observation, ELISA analysis demonstrated that ODNs A151 suppressed Th1 cytokines IFN-gamma and TNF-alpha, and augmented Th2 cytokines IL-4 and IL-10 in vitro. This study provides the first experimental evidence that suppressive ODNs inhibit the development of atherosclerosis through inhibition of the STAT1/4 and T-bet pathways, which further modulate the Th1/Th2 balance in vivo.
动脉粥样硬化是一种慢性炎症性疾病，分别受到辅助性T（Th）1和Th2淋巴细胞的正向和负向调节。最近的研究结果表明，表达TTAGGG基序的抑制性寡脱氧核苷酸（ODN）可选择性降低Th1细胞因子的产生，并已被证明在阻断器官特异性自身免疫性疾病的发展方面有效。在当前的研究中，我们假设抑制性ODN可能会改变动脉粥样硬化的发展。将300微克的ODN A151（TTAGGG）或非特异性ODN 1612注射到8周龄的纯合ApoE（-/-）雄性小鼠体内。ODN A151可显著减小动脉粥样硬化病变大小，而非特异性ODN则无此作用。抑制性ODN A151可显著减弱动脉粥样硬化形成过程中的关键炎症因子MCP-1和VCAM-1。在脾淋巴细胞中，流式细胞术分析显示ODN A151降低了产生IFN-γ的Th1细胞百分比，并略微增加了产生IL-4的Th2细胞百分比，表明抑制性ODN在体内使Th1/Th2平衡向Th2炎症倾斜。此外，ODN A151下调了STAT1和STAT4的磷酸化，并抑制了Th1的信号调节剂T-bet的上调，且不影响与Th2表型相关的GATA-3和STAT6。与这一体内观察结果一致，ELISA分析表明ODN A151在体外抑制了Th1细胞因子IFN-γ和TNF-α，并增强了Th2细胞因子IL-4和IL-10。本研究提供了首个实验证据，即抑制性ODN通过抑制STAT1/4和T-bet途径来抑制动脉粥样硬化的发展，这进一步调节了体内的Th1/Th2平衡。
