Tirumuru Nagaraja, Pretto Carla D, Castro Jorge Luiza A, Spindler Katherine R
Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan, USA.
mSphere. 2016 Apr 20;1(2). doi: 10.1128/mSphere.00079-16. eCollection 2016 Mar-Apr.
Mouse adenovirus type 1 (MAV-1) infects endothelial cells and disrupts the blood-brain barrier (BBB), causing encephalitis in inbred and outbred mice. Using a virus mutant that does not produce the early region 1A protein E1A, we investigated whether the activity of this known viral transcriptional regulator is needed for BBB disruption and other phenotypes associated with encephalitis. The wild-type (wt) virus and E1A mutant virus caused similar levels of permeability of sodium fluorescein in brains of infected mice. In an in vitro assay of BBB integrity, wt and mutant virus caused similar decreases in transendothelial electrical resistance in primary mouse brain endothelial cell monolayers. These results indicate that E1A protein does not contribute to disruption of BBB integrity in animals or cultured cells. Both wt and E1A mutant virus infection of mice led to similar increases in the activity of two matrix metalloproteinases known to correlate with BBB disruption, MMP2 and MMP9, while causing no increase in the steady-state expression of MMP2 or MMP9 mRNA. In contrast, the amount of MMP3 transcripts increased upon infection by both viruses and to a higher level in infections by the mutant virus lacking E1A protein production. There was no difference in the levels of steady-state expression of mRNA for tight junction proteins among mock virus, wt virus, and mutant virus infections. Thus, the MAV-1 E1A protein does not measurably affect BBB integrity in the parameters assayed, although it reduces the amount of MMP3 mRNA steady-state expression induced in brains upon infection. IMPORTANCE Encephalitis can be caused by viruses, and it is potentially life-threatening because of the vital nature of the brain and the lack of treatment options. MAV-1 produces viral encephalitis in its natural host, providing a model for investigating factors involved in development of encephalitis. MAV-1 infection disrupts the BBB and increases activity of matrix metalloproteinases in brains of infected mice. We investigated whether the major transcriptional regulator of adenoviruses, E1A protein, is responsible for any of the specific phenotypes that result from MAV-1 infection. For some of the functions assayed, an E1A mutant virus behaved like wild-type virus. However, expression of mRNA for one matrix metalloproteinase was higher in the virus lacking E1A protein production. This highlights the complex nature of encephalitis and suggests that E1A may have transcriptional effects on host genes important for the development of encephalitis.
1型小鼠腺病毒(MAV-1)感染内皮细胞并破坏血脑屏障(BBB),在近交系和远交系小鼠中引发脑炎。我们使用一种不产生早期区域1A蛋白E1A的病毒突变体,研究了这种已知的病毒转录调节因子的活性对于破坏血脑屏障以及与脑炎相关的其他表型是否必要。野生型(wt)病毒和E1A突变病毒在感染小鼠的脑中引起相似水平的荧光素钠通透性。在一项血脑屏障完整性的体外测定中,wt病毒和突变病毒在原代小鼠脑内皮细胞单层中引起相似程度的跨内皮电阻降低。这些结果表明,E1A蛋白对动物或培养细胞中血脑屏障完整性的破坏没有作用。wt病毒和E1A突变病毒感染小鼠均导致已知与血脑屏障破坏相关的两种基质金属蛋白酶MMP2和MMP9的活性出现相似程度的增加,而MMP2或MMP9 mRNA的稳态表达没有增加。相反,两种病毒感染后MMP3转录本的量均增加,且在缺乏E1A蛋白产生的突变病毒感染中增加到更高水平。在模拟病毒、wt病毒和突变病毒感染之间,紧密连接蛋白mRNA的稳态表达水平没有差异。因此,MAV-1 E1A蛋白在所检测的参数中对血脑屏障完整性没有可测量的影响,尽管它降低了感染后脑内诱导的MMP3 mRNA稳态表达量。重要性 脑炎可由病毒引起,由于脑的重要性质和缺乏治疗选择,它具有潜在的生命威胁性。MAV-1在其自然宿主中产生病毒性脑炎,为研究参与脑炎发展的因素提供了一个模型。MAV-1感染破坏血脑屏障并增加感染小鼠脑中基质金属蛋白酶的活性。我们研究了腺病毒的主要转录调节因子E1A蛋白是否对MAV-1感染导致的任何特定表型负责。对于所检测的一些功能,E1A突变病毒的表现与野生型病毒相似。然而,在缺乏E1A蛋白产生的病毒中,一种基质金属蛋白酶的mRNA表达更高。这突出了脑炎的复杂性,并表明E1A可能对脑炎发展重要的宿主基因具有转录作用。