TNF-alpha After Anterior Chamber Inoculation of HSV-1

描述（由申请方提供）：用HSV-1的科斯菌株对正常胸腺BALB/c小鼠进行前房（AC）接种仅导致未注射眼的视网膜炎。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。在将HSV-1 AC接种到一只眼睛中后，病毒通过从同侧SCN扩散进入对侧眼睛的视神经和视网膜。虽然病毒感染对侧SCN，但它不会从那里传播到注射眼的视神经和视网膜。如果病毒可以包含在SCN中，则这种包含可能会防止未注射眼睛的视网膜的病毒感染。我的初步研究表明，肿瘤坏死因子-α（TNF-α），一种促炎细胞因子，在SCN中的生产是重要的限制病毒传播，并可能保留视网膜的注射眼。使用免疫组织化学、免疫荧光、空斑测定和分子克隆方法，我计划：（1）通过阻断TNF-α来确定TNF-α在HSV-1感染的BALB/c小鼠的眼和脑中的作用，（2）鉴定SCN和注射眼中产生TNF-α的细胞，和（3）构建可以被诱导表达TNF-α的HSV-1的无缺陷重组体。这些研究的结果将进一步了解TNF-α在病毒感染期间在中枢神经系统中的作用，并可能为开发一种基于奎宁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.