Kaneyama Tomoki, Tomiki Hiroki, Tsugane Sayaka, Inaba Yuji, Ichikawa Motoki, Akiba Hisaya, Yagita Hideo, Kim Byung S, Koh Chang-Sung
Department of Biomedical Laboratory Sciences, Graduate School of Medicine, Shinshu University, Matsumoto, Nagano 390-8621, Japan Research Fellow of the Japan Society for the Promotion of Science, Chiyoda-ku, Tokyo 102-0083, Japan.
Department of Biomedical Laboratory Sciences, Graduate School of Medicine, Shinshu University, Matsumoto, Nagano 390-8621, Japan.
Int Immunol. 2014 Jul;26(7):369-81. doi: 10.1093/intimm/dxt056. Epub 2014 Jan 31.
Infection by Theiler's murine encephalomyelitis virus (TMEV) in the central nervous system (CNS) induces an immune-mediated demyelinating disease in susceptible mouse strains and serves as a relevant infection model for human multiple sclerosis. T-cell immunoglobulin and mucin domain-3 (TIM-3) has been demonstrated to play a crucial role in the maintenance of peripheral tolerance. In this study, we examined the regulatory role of the TIM-3 pathway in the development of TMEV-induced demyelinating disease (TMEV-IDD). The expression of TIM-3 was increased at both protein and mRNA levels in the spinal cords of mice with TMEV-IDD compared with naive controls. In addition, by utilizing a blocking mAb, we demonstrate that TIM-3 negatively regulates TMEV-specific ex vivo production of IFN-γ and IL-10 by CD4(+) T cells and IFN-γ by CD8(+) T cells from the CNS of mice with TMEV-IDD at 36 days post-infection (dpi). In vivo blockade of TIM-3 by using the anti-TIM-3 mAb resulted in significant exacerbation of the development of TMEV-IDD both clinically and histologically. The number of infiltrating mononuclear cells in the CNS was also increased in mice administered with anti-TIM-3 mAb both at the induction phase (10 dpi) and at the effector phase (36 dpi). Flow cytometric analysis of intracellular cytokines revealed that the number of CD4(+) T cells producing TNF, IL-4, IL-10 and IL-17 was significantly increased at the effector phase in the CNS of anti-TIM-3 mAb-treated mice. These results suggest that the TIM-3 pathway plays a critical role in the regulation of TMEV-IDD.
在中枢神经系统(CNS)中,泰勒氏鼠脑脊髓炎病毒(TMEV)感染会在易感小鼠品系中引发免疫介导的脱髓鞘疾病,并作为人类多发性硬化症的相关感染模型。T细胞免疫球蛋白和粘蛋白结构域3(TIM-3)已被证明在维持外周免疫耐受中起关键作用。在本研究中,我们研究了TIM-3通路在TMEV诱导的脱髓鞘疾病(TMEV-IDD)发展中的调节作用。与未感染的对照组相比,TMEV-IDD小鼠脊髓中TIM-3的蛋白和mRNA水平均升高。此外,通过使用阻断性单克隆抗体,我们证明在感染后36天(dpi),TIM-3负向调节TMEV特异性的体外IFN-γ和IL-10的产生,以及TMEV-IDD小鼠中枢神经系统中CD8(+)T细胞产生的IFN-γ。使用抗TIM-3单克隆抗体在体内阻断TIM-3会导致TMEV-IDD在临床和组织学上的发展显著加剧。在诱导期(10 dpi)和效应期(36 dpi)给予抗TIM-3单克隆抗体的小鼠中枢神经系统中浸润的单核细胞数量也增加。细胞内细胞因子的流式细胞术分析显示,在效应期,抗TIM-3单克隆抗体处理的小鼠中枢神经系统中产生TNF、IL-4、IL-10和IL-17的CD4(+)T细胞数量显著增加。这些结果表明,TIM-3通路在TMEV-IDD的调节中起关键作用。