单纯疱疹病毒1型(HSV-1)是一种广泛的传染性病原体,大多在粘膜进入侧引起轻微症状。然而,系统分布,特别是在免疫受损患者中病毒重新激活后,可能引发先天免疫反应并诱导器官损伤。在这些条件下,HSV-1可能感染血管内皮细胞,但对这些细胞中HSV-1复制的调节和可能的防御机制知之甚少。当前的研究解决了宿主细胞蛋白AMP激活的蛋白激酶(AMPK)一个重要的代谢传感器,可以控制HSV-1在内皮细胞中的复制。我们表明,通过TCID50滴定监测,AMPKα1和/或AMPKα2的催化亚基下调增加了HSV-1的复制,虽然是一种有效的AMPK激动剂,MK-8722强烈抑制了它。MK-8722诱导AMPK下游靶乙酰辅酶A羧化酶(ACC)和mTOR(Raptor)的雷帕霉素敏感衔接蛋白的持续磷酸化,与此相关,ACC1介导的脂质合成的损害和雷帕霉素复合物-1(mTORC1)途径的机制靶标。由于Torin-2对mTOR的阻断以及siRNA对ACC1的下调也降低了HSV-1的复制,MK-8722可能通过mTORC1和ACC1抑制发挥其抗病毒作用。重要的是,即使在HSV-1之后添加MK-8722也能够减少病毒复制。一起,我们的数据强调了在全身感染时内皮细胞作为HSV-1复制的宿主细胞的重要性,并鉴定了AMPK,代谢宿主细胞蛋白,作为抗HSV-1感染及其严重后果的抗病毒策略的潜在目标。重要性单纯疱疹病毒1型(HSV-1)是一种常见的病原体,可导致人类水疱或唇疱疹。它在受感染的个体中仍然潜伏,可以多次重新激活。在不利条件下,例如,在免疫功能低下的患者中,HSV-1可导致严重的并发症,如脑炎,脑膜炎,或失明。在这些情况下,血管表面内皮细胞感染可能导致疾病的表现。这里,我们描述了AMP激活的蛋白激酶(AMPK)的作用,细胞能量代谢的有效调节剂,在内皮细胞中HSV-1复制。虽然AMPK的下调增强了HSV-1的复制,药理学AMPK激活通过限制所需宿主细胞大分子如蛋白质或脂肪酸的可用性来抑制它。这些数据突出了代谢宿主细胞蛋白作为抗病毒靶标的作用,并揭示了内皮AMPK的激活作为保护免受HSV-1感染严重后果的潜在策略。
Herpes simplex virus type 1 (HSV-1) is a widespread contagious pathogen, mostly causing mild symptoms on the mucosal entry side. However, systemic distribution, in particular upon reactivation of the virus in immunocompromised patients, may trigger an innate immune response and induce damage of organs. In these conditions, HSV-1 may infect vascular endothelial cells, but little is known about the regulation of HSV-1 replication and possible defense mechanisms in these cells. The current study addresses the question of whether the host cell protein AMP-activated protein kinase (AMPK), an important metabolic sensor, can control HSV-1 replication in endothelial cells. We show that downregulation of the catalytic subunits AMPKα1 and/or AMPKα2 increased HSV-1 replication as monitored by TCID50 titrations, while a potent AMPK agonist, MK-8722, strongly inhibited it. MK-8722 induced a persistent phosphorylation of the AMPK downstream targets acetyl-CoA carboxylase (ACC) and the rapamycin-sensitive adaptor protein of mTOR (Raptor) and, related to this, impairment of ACC1-mediated lipid synthesis and the mechanistic target of the rapamycin complex-1 (mTORC1) pathway. Since blockade of mTOR by Torin-2 as well as downregulation of ACC1 by siRNA also decreased HSV-1 replication, MK-8722 is likely to exert its anti-viral effect via mTORC1 and ACC1 inhibition. Importantly, MK-8722 was able to reduce virus replication even when added after HSV-1. Together, our data highlight the importance of endothelial cells as host cells for HSV-1 replication upon systemic infection and identify AMPK, a metabolic host cell protein, as a potential target for antiviral strategies against HSV-1 infection and its severe consequences. IMPORTANCE Herpes simplex virus type 1 (HSV-1) is a common pathogen that causes blisters or cold sores in humans. It remains latent in infected individuals and can be reactivated multiple times. In adverse conditions, for instance, in immunocompromised patients, HSV-1 can lead to serious complications such as encephalitis, meningitis, or blindness. In these situations, infection of endothelial cells lining the surface of blood vessels may contribute to the manifestation of disease. Here, we describe the role of AMP-activated protein kinase (AMPK), a potent regulator of cellular energy metabolism, in HSV-1 replication in endothelial cells. While downregulation of AMPK potentiates HSV-1 replication, pharmacological AMPK activation inhibits it by limiting the availability of required host cell macromolecules such as proteins or fatty acids. These data highlight the role of metabolic host cell proteins as antiviral targets and reveal activation of endothelial AMPK as a potential strategy to protect from severe consequences of HSV-1 infection.