Kumnok Jirawat, Ulrich Reiner, Wewetzer Konstantin, Rohn Karl, Hansmann Florian, Baumgartner Wolfgang, Alldinger Susanne
Department of Pathology, University of Veterinary Medicine, Hannover, Germany.
J Neurovirol. 2008 May;14(3):205-17. doi: 10.1080/13550280802008305.
The BeAn strain of Theiler's murine encephalomyelitis virus (TMEV) induces demyelinating disease in susceptible mice comparable to human multiple sclerosis. Recent in vivo studies showed that matrix metalloproteinases (MMPs) and their inhibitors (tissue inhibitors of MMPs, TIMPs) are associated with demyelination in Theiler's murine encephalomyelitis. The present study was performed to evaluate the in vitro MMP and TIMP expression in astrocytes and microglia following TMEV infection. Brain cell cultures from SJL/J mice were infected with the BeAn strain of TMEV and the expressions of 11 MMPs and 4 TIMPs were evaluated by reverse-transcription quantitative polymerase chain reaction (RT-qPCR) at different time points post infection (p.i.). In control astrocytes and microglia, a constitutive expression of MMP-2, -3, -9, -10, -12, -13, -14, -15, -24 and TIMP-2 to -4 was detected. In addition, TIMP-1 and MMP-11 was found in astrocytes only, and MMP-7 was absent in both cells cultures. RT-qPCR demonstrated high virus RNA copy numbers in astrocytes and a low amount in microglia. In accordance, TMEV antigen was detected in astrocytes, whereas it was below the limit of detection in microglia. MMP-3, -9, -10, -12, and -13 as well as TIMP-1 were the enzymes most prominently up-regulated in TMEV-infected astrocytes. In contrast, TMEV infection was associated with a down-regulation of MMPs and TIMPs in microglia. Conclusively, in addition to inflammatory infiltrates, TMEV-induced astrocytic MMPs might trigger a proteolysis cascade leading to an opening of the blood-brain barrier and demyelination in vivo.
泰勒鼠脑脊髓炎病毒(TMEV)的BeAn株可在易感小鼠中诱发脱髓鞘疾病,与人类多发性硬化症相似。最近的体内研究表明,基质金属蛋白酶(MMPs)及其抑制剂(MMPs组织抑制剂,TIMPs)与泰勒鼠脑脊髓炎中的脱髓鞘有关。本研究旨在评估TMEV感染后星形胶质细胞和小胶质细胞中MMP和TIMP的体外表达。用TMEV的BeAn株感染SJL/J小鼠的脑细胞培养物,并在感染后(p.i.)的不同时间点通过逆转录定量聚合酶链反应(RT-qPCR)评估11种MMPs和4种TIMPs的表达。在对照星形胶质细胞和小胶质细胞中,检测到MMP-2、-3、-9、-10、-12、-13、-14、-15、-24和TIMP-2至-4的组成性表达。此外,仅在星形胶质细胞中发现TIMP-1和MMP-11,两种细胞培养物中均未发现MMP-7。RT-qPCR显示星形胶质细胞中病毒RNA拷贝数高,小胶质细胞中病毒RNA拷贝数低。相应地,在星形胶质细胞中检测到TMEV抗原,而在小胶质细胞中低于检测限。MMP-3、-9、-10、-12和-13以及TIMP-1是TMEV感染的星形胶质细胞中上调最显著的酶。相反,TMEV感染与小胶质细胞中MMPs和TIMPs的下调有关。总之,除了炎症浸润外,TMEV诱导的星形胶质细胞MMPs可能引发蛋白水解级联反应,导致体内血脑屏障开放和脱髓鞘。