Zhou Xiangyu, Li Xiaoyong, Feng Meina, Zhang Qi, Yang Zhendong
Department of Neurology, The Affiliated First Hospital of Shizuishan, The Ningxia Medical University, Shizuishan, Ningxia 753200, P.R. China.
Department of Pathology, Medical College, Hubei Polytechnic University, Huangshi, Hubei 435003, P.R. China.
Mol Med Rep. 2015 Nov;12(5):7425-32. doi: 10.3892/mmr.2015.4358. Epub 2015 Sep 24.
The aim of the present study was to investigate the possible role of cytotoxic T lymphocytes (CTL) and mononuclear macrophages in the pathogenic processes of experimental animals. To construct a chronic experimental allergic encephalomyelitis (EAE) model, an artificially synthesized myelin oligodendrocyte glycoprotein (MOG)35‑55 peptide was used to induce C57BL/6 mice. Subsequently, the experimental animals were investigated at the level of their nervous function, and histopathological, immunohistochemical and fluorescence immunohistochemical experiments were performed at different time points following immunization. The expression of immune molecules and cytokines associated with the activation of the mononuclear macrophages and CTL during the different stages was assessed by western blotting and reverse transcription‑quantitative polymerase chain reaction. As a result, the MOG35‑55 peptide was identified as being successful at inducing C57BL/6 mice for the development of the EAE model. A modest level of mononuclear macrophage and lymphocyte infiltration was observed in the central nervous system (CNS), although no infiltration of neutrophils was observed. A sporadic flaky deletion of the myelin sheath was also identified. The activation and proliferation of mononuclear macrophages, including microglia cells, was clearly demonstrated. Furthermore, the expression levels of major histocompatibility complex class I and II molecules and interleukin‑12 in the brain, which is associated with the activation and proliferation of mononuclear macrophages, increased over the duration of the experiment compared with less pronounced changes in the expression levels of interferon (IFN)‑γ, Fas and perforin in the CNS, which are associated with the function of CTL. The secretion of IFN‑γ in the spleen increased during the morbidity peak, however, any noticeable activation and proliferation of CD8+ T cells was absent. These results demonstrated that the induced immune response mediated by mononuclear macrophages made a more important contribution compared with CTL towards the pathological process of myelin sheath injury. Mononuclear macrophages are therefore, identified as being one of the most significant effector cell types to directly injure the myelin sheath in the CNS.
本研究的目的是探讨细胞毒性T淋巴细胞(CTL)和单核巨噬细胞在实验动物致病过程中的可能作用。为构建慢性实验性变应性脑脊髓炎(EAE)模型,使用人工合成的髓鞘少突胶质细胞糖蛋白(MOG)35-55肽诱导C57BL/6小鼠。随后,对实验动物的神经功能水平进行研究,并在免疫后的不同时间点进行组织病理学、免疫组织化学和荧光免疫组织化学实验。通过蛋白质印迹法和逆转录-定量聚合酶链反应评估不同阶段与单核巨噬细胞和CTL激活相关的免疫分子和细胞因子表达。结果表明,MOG35-55肽成功诱导C57BL/6小鼠发生EAE模型。在中枢神经系统(CNS)中观察到适度水平的单核巨噬细胞和淋巴细胞浸润,尽管未观察到中性粒细胞浸润。还发现髓鞘有散在片状缺失。明显证实了包括小胶质细胞在内的单核巨噬细胞的激活和增殖。此外,与单核巨噬细胞激活和增殖相关的脑内主要组织相容性复合体I类和II类分子以及白细胞介素-12的表达水平在实验过程中升高,而与CTL功能相关的中枢神经系统中干扰素(IFN)-γ、Fas和穿孔素的表达水平变化不明显。发病高峰期脾脏中IFN-γ的分泌增加,然而,未观察到CD8+T细胞有明显的激活和增殖。这些结果表明,与CTL相比,单核巨噬细胞介导的诱导免疫反应对髓鞘损伤的病理过程贡献更大。因此,单核巨噬细胞被确定为直接损伤中枢神经系统髓鞘的最重要效应细胞类型之一。