TNF-alpha After Anterior Chamber Inoculation of HSV-1

描述（由申请人提供）：前房（AC）接种无菌BALB/c小鼠与HSV-1的KOS株只会导致未注射眼睛的视网膜炎。病毒进入下丘脑视交叉上核（SCN）的时间似乎是决定小鼠是否发生双侧视网膜炎的关键。单眼AC接种HSV-1后，病毒从同侧SCN扩散进入对侧眼视神经和视网膜。虽然病毒感染了对侧SCN，但它不会从那里扩散到被注射眼睛的视神经和视网膜。如果病毒可以被控制在SCN中，这种控制可能会防止病毒感染未注射眼睛的视网膜。我的初步研究表明，在SCN中产生肿瘤坏死因子- α (tnf - α)，一种促炎细胞因子，对于限制病毒传播和可能保留注射眼睛的视网膜很重要。利用免疫组织化学、免疫荧光、斑块测定和分子克隆方法，我计划：(1)通过阻断TNF-a来确定tnf - α在HSV-1感染的BALB/c小鼠的眼睛和大脑中的作用，(2)鉴定SCN和注射的眼睛中产生tnf - α的细胞，(3)构建一个可以诱导表达tnf - α的无缺陷重组HSV-1。这些研究结果将进一步了解病毒感染期间中枢神经系统中tnf - α的作用，并可能为开发基于细胞因子的抗病毒疗法以帮助免疫系统早期清除病毒打开大门。

DESCRIPTION (provided by applicant): Anterior chamber (AC) inoculation of euthymic BALB/c mice with the KOS strain of HSV-1 results in retinitis of the uninjected eye only. Timing of virus entry into the suprachiasmatic nucleus (SCN) of the hypothalamus appears to be crucial in determining whether mice develop bilateral retinitis. After AC inoculation of HSV-1 into one eye, virus enters the optic nerve and retina of the contralateral eye by spreading from the ipsilateral SCN. Although virus infects the contralateral SCN, it does not spread from there to the optic nerve and retina of the injected eye. If virus can be contained in the SCN, such containment would likely prevent virus infection of the retina of the uninjected eye. My preliminary studies suggest that production of tumor necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine, in the SCN is important for limitation of virus spread and possibly for sparing of the retina of the injected eye. Using immunohistochemistry, immunofluorescence, plaque assays, and molecular cloning methods, I plan to: (1) determine the role of TNF-alpha in the eyes and brain of HSV-1 infected BALB/c mice by blocking TNF-a, (2) identify the cells in the SCN and in the injected eye that produce TNF-alpha, and (3) construct a non-defective recombinant of HSV-1 that can be induced to express TNF-alpha. The results of these studies will provide a further understanding of the role of TNF-alpha in the central nervous system during viral infection, and perhaps open the door for the development of a cytokine-based anti-viral therapy to assist the immune system in early clearance of virus.