Role of macrophages in control of ocular HSV

HSV-1感染在美国非常频繁，是病毒致盲的主要原因。角膜 HSV-1引起的损伤似乎主要是由免疫反应介导的。我们发现巨噬细胞 在眼部感染HSV-1的小鼠的角膜中形成早期和主要的浸润物。我们还展示了 通过注射集落刺激因子-1(CSF-1)优先扩增M2巨噬细胞亚群。 1)减少了眼部病毒的复制和潜伏期重新激活，而M1亚群的激活是促进的 炎症性和加重的眼病。眼部极早期浸润性病变的自然历史分析 感染的小鼠提示M1和M2巨噬细胞的角膜渗透模式存在二分法，即 在时间上与随后的其他免疫反应有关，病毒从 角膜，并建立潜伏期。这些结果为区分免疫反应提供了一个框架。 急性和潜伏性HSV-1感染的介导性加重与免疫介导性控制。基于我们的 根据已公布的初步数据，我们的主要假设是巨噬细胞激活的自然变异 对于M1或M2疾病相关的表型在决定是否诱发炎症方面起着关键作用 疾病和病毒复制。因此，可以使用对M1和M2巨噬细胞隔室的操纵 以保护眼球前段的完整性，包括角膜，以应对感染。 具体地说，我们将测试巨噬细胞亚群中自噬的操纵是否可以用来控制 眼部单纯疱疹病毒1型感染。拟议研究的可行性植根于我们的战略，利用有条件的 我们已经建立的基因敲除小鼠直接评估体内M1和M2功能与病毒有关 眼部复制、眼部疾病和眼部感染小鼠潜伏期再激活的建立。我们会： (1)检测眼部感染小鼠角膜巨噬细胞对M2的激活表型是否发生改变 将导致减少原发感染、炎症反应和眼病，并缩短潜伏期- 重新激活；以及(2)测试抑制M2巨噬细胞的自噬是否会增加眼病和潜伏期- 重新激活，而在M1巨噬细胞中抑制它可以减少炎症、眼病和潜伏期- 重新激活。我们将确定在转基因小鼠中阻断自噬的影响 单纯疱疹病毒1型(即γ34.5)在M1(NOS2)或M2(Arg1)启动子下的自噬基因 WT型单纯疱疹病毒McKrae株或单纯疱疹病毒1型McKrae株的γ34.5缺失突变体。在这两个目标上，我们将进一步测试 抗病毒药物定量方面改善疾病进程的相关机制 巨噬细胞的反应、吞噬和/或自噬以及对其他免疫浸润物和 细胞因子的释放。

HSV-1 infections are very frequent in the U.S. and are a major cause of viral-induced blindness. The corneal damage induced by HSV-1 appears to be mediated primarily by immune responses. We found that macrophages form early and predominant infiltrates in the corneas of mice ocularly infected with HSV-1. We have also shown that preferential expansion of the M2 macrophage subpopulation by injection of colony stimulating factor-1 (CSF- 1) reduced ocular virus replication and latency-reactivation, whereas activation of the M1 subpopulation was pro- inflammatory and exacerbated eye disease. Analyses of the natural history of very early infiltrates in ocularly infected mice suggest a dichotomy in the patterns of corneal infiltration by M1 and M2 macrophages that is temporally associated with subsequent involvement of other immune responses, clearance of the virus from the cornea, and establishment of latency. These results provide a framework for differentiating immune response- mediated exacerbation vs. immune-mediated control of acute and latent HSV-1 infections. Based on our published and preliminary data, our main hypothesis is that the natural variation in the activation of macrophages towards the M1 or M2 disease-relevant phenotype plays a key role in determining induction of inflammation, eye disease, and virus replication. Therefore, manipulation of M1 and M2 macrophage compartments can be used to safeguard the integrity of the anterior segment of the eye including the cornea in response to infection. Specifically, we will test whether manipulation of autophagy in the macrophage subsets can be used to control ocular HSV-1 infection. The feasibility of the proposed studies is rooted in our strategy that utilizes conditional knockout mice that we have generated to directly evaluate the M1 and M2 functions in vivo with regards to virus replication in the eye, eye disease and establishment of latency-reactivation in ocularly infected mice. We will: (1) Test if altering the phenotype of macrophage activation towards M2 in the cornea of ocularly infected mice will lead to a reduction in primary infection, inflammatory responses and eye disease, and a reduction in latency- reactivation; and (2) Test if inhibition of autophagy in M2 macrophages enhances eye disease and latency- reactivation, while its inhibition in M1 macrophages decreases inflammation, eye disease and latency- reactivation. We will determine the impact of blocking autophagy in transgenic mice expressing the anti- autophagy gene of HSV-1 (i.e., γ34.5) under the M1 (NOS2) or the M2 (Arg1) promoters following infection with WT HSV-1 strain McKrae or a γ34.5 deletion mutant of HSV-1 strain McKrae. In both Aims, we will further test the mechanisms associated with amelioration of the disease process in terms of quantification of antiviral responses, phagocytosis and/or autophagy in the macrophages and the impact on other immune infiltrates and cytokine release.