Role of macrophages in control of ocular HSV

巨噬细胞在控制眼部 HSV 中的作用

• 批准号: 9759926
• 负责人: HOMAYON GHIASI
• 金额: $ 42.5万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9759926
• 关键词: Acute; Anterior eyeball segment structure; Antiviral Agents; Antiviral Response; Apoptosis; Autophagocytosis; Blindness; Cells; Characteristics; Cornea; Data; Development; Disease; Dose; Exhibits; Eye; Eye Infections; Eye diseases; Frequencies; Genes; Herpetic Keratitis; Homeostasis; Immune; Immune response; Immune system; Infection; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Injections; Innate Immune Response; Kinetics; Knockout Mice; Latent Virus; Lead; Leucocytic infiltrate; Macrophage Activation; Macrophage Colony-Stimulating Factor; Maintenance; Mediating; Mononuclear; Mouse Strains; Mus; Mutate; NOS2A gene; Natural History; Pathway interactions; Pattern; Phagocytes; Phagocytosis; Phenotype; Plant Roots; Play; Predisposition; Primary Infection; Process; Publishing; Reagent; Recurrence; Resolution; Role; STAT1 gene; Simplexvirus; Testing; Therapeutic; Time; Tissues; Transgenic Mice; Tumor-infiltrating immune cells; Variant; Viral; Viral Eye Infections; Viral Load result; Virus; Virus Diseases; Virus Latency; Virus Replication; Vision; base; clinically relevant; clinically significant; cytokine; in vivo; inhibition of autophagy; macrophage; mouse model; mutant; novel; preservation; promoter; reactivation from latency; response; transcription factor

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.

HSV-1 感染在美国非常频繁，是病毒性失明的主要原因。 HSV-1 引起的损伤似乎主要是由免疫反应介导的。 我们还表明，在眼部感染 HSV-1 的小鼠的角膜中形成早期和主要的浸润。 通过注射集落刺激因子-1（CSF- 1) 减少眼部病毒复制和潜伏期再激活，而 M1 亚群的激活则促进 眼部早期浸润的自然史分析。 受感染的小鼠表明 M1 和 M2 巨噬细胞的角膜浸润模式存在二分法，即 暂时与随后涉及的其他免疫反应、病毒从机体中的清除相关 角膜和潜伏期的建立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 感染的介导的恶化与免疫介导的控制基于我们的研究。 已发表的初步数据，我们的主要假设是巨噬细胞激活的自然变异 M1 或 M2 疾病相关表型在决定炎症、眼部疾病的诱导中起着关键作用 因此，可以使用 M1 和 M2 巨噬细胞区室的操作。 保护眼前段（包括角膜）的完整性，以应对感染。 具体来说，我们将测试巨噬细胞亚群中自噬的操纵是否可以用于控制 所提出的研究的可行性植根于我们利用条件的策略。 我们培育的基因敲除小鼠可直接评估 M1 和 M2 体内针对病毒的功能 眼部复制、眼部疾病以及眼部感染小鼠潜伏期再激活的建立我们将： (1) 测试眼部感染小鼠角膜中巨噬细胞活化表型是否改变为M2 将导致原发感染、炎症反应和眼部疾病的减少，以及潜伏期的减少- 重新激活；(2) 测试 M2 巨噬细胞自噬的抑制是否会加剧眼部疾病和潜伏期 重新激活，而其对 M1 巨噬细胞的抑制可减少炎症、眼部疾病和潜伏期 - 我们将确定阻断自噬对表达抗-的转基因小鼠的影响。 感染后 M1 (NOS2) 或 M2 (Arg1) 启动子下的 HSV-1 自噬基因（即 γ34.5） WT HSV-1 McKrae 株或 HSV-1 McKrae 株的 γ34.5 缺失突变体 在这两个目标中，我们将进一步测试。 抗病毒药物定量方面与疾病过程改善相关的机制 巨噬细胞中的反应、吞噬作用和/或自噬以及对其他免疫浸润的影响和 细胞因子释放。