TNF-alpha After Anterior Chamber Inoculation of HSV-1

描述(由申请人提供)：前房(AC)接种Kos株HSV-1的BALB/c小鼠只会导致未注射的眼睛发生视网膜炎。病毒进入下丘脑视交叉上核(SCN)的时间似乎对确定小鼠是否发展为双侧视网膜炎至关重要。将HSV-1 AC接种到一只眼内后，病毒通过同侧SCN传播进入对侧的视神经和视网膜。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。如果病毒能够被控制在SCN中，这种遏制很可能会防止病毒感染未注射的眼睛的视网膜。我的初步研究表明，在SCN中产生的肿瘤坏死因子-α(TNF-α)是一种促炎细胞因子，对于限制病毒传播以及可能对保留注射眼的视网膜至关重要。利用免疫组织化学、免疫荧光、空斑分析和分子克隆方法，我计划：(1)通过阻断肿瘤坏死因子-α，确定肿瘤坏死因子-α在单纯疱疹病毒1型感染的BALB/c小鼠眼和脑中的作用；(2)鉴定脊髓核内和注射眼内产生肿瘤坏死因子-α的细胞；(3)构建能够诱导表达肿瘤坏死因子-α的无缺陷的单纯疱疹病毒1型重组体。这些研究结果将进一步了解肿瘤坏死因子-α在病毒感染过程中在中枢神经系统中的作用，并可能为以细胞因子为基础的抗病毒治疗的发展打开大门，以帮助免疫系统早期清除病毒。

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.