Department of Medicine Huddinge, Center for Infectious Medicine, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden.
PLoS Pathog. 2020 Feb 7;16(2):e1008297. doi: 10.1371/journal.ppat.1008297. eCollection 2020 Feb.
Hantaviruses, zoonotic RNA viruses belonging to the order Bunyavirales, cause two severe acute diseases in humans, hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). Hantavirus-infected patients show strong cytotoxic lymphocyte responses and hyperinflammation; however, infected cells remain mostly intact. Hantaviruses were recently shown to inhibit apoptosis in infected cells. By inhibiting granzyme B- and TRAIL-mediated apoptosis, hantaviruses specifically and efficiently inhibit cytotoxic lymphocyte-mediated killing of infected cells. Hantaviruses also strongly inhibit apoptosis triggered intrinsically; i.e., initiated through intracellular activation pathways different from those used by cytotoxic lymphocytes. However, insights into the latter mechanisms are currently largely unknown. Here, we dissected the mechanism behind how hantavirus infection, represented by the HFRS-causing Hantaan virus and the HPS-causing Andes virus, results in resistance to staurosporine-induced apoptosis. Less active caspase-8 and caspase-9, and consequently less active caspase-3, was observed in infected compared to uninfected staurosporine-exposed cells. While staurosporine-exposed uninfected cells showed massive release of pro-apoptotic cytochrome C into the cytosol, this was not observed in infected cells. Further, hantaviruses prevented activation of BAX and mitochondrial outer membrane permeabilization (MOMP). In parallel, a significant increase in levels of the pro-survival factor BCL-2 was observed in hantavirus-infected cells. Importantly, direct inhibition of BCL-2 by the inhibitor ABT-737, as well as silencing of BCL-2 by siRNA, resulted in apoptosis in staurosporine-exposed hantavirus-infected cells. Overall, we here provide a tentative mechanism by which hantaviruses protect infected cells from intrinsic apoptosis at the mitochondrial level by inducing an increased expression of the pro-survival factor BCL-2, thereby preventing MOMPs and subsequent activation of caspases. The variety of mechanisms used by hantaviruses to ensure survival of infected cells likely contribute to the persistent infection in natural hosts and may play a role in immunopathogenesis of HFRS and HPS in humans.
汉坦病毒是布尼亚病毒目(order Bunyavirales)中的一种正链 RNA 病毒,会导致人类罹患两种严重的急性疾病,即肾综合征出血热(HFRS)和汉坦病毒肺综合征(HPS)。汉坦病毒感染患者会表现出强烈的细胞毒性淋巴细胞反应和过度炎症;然而,受感染的细胞大多保持完整。最近发现汉坦病毒可以抑制感染细胞的细胞凋亡。汉坦病毒通过抑制颗粒酶 B 和 TRAIL 介导的细胞凋亡,特异性且有效地抑制细胞毒性淋巴细胞介导的感染细胞杀伤。汉坦病毒还强烈抑制内在触发的凋亡;即,通过与细胞毒性淋巴细胞不同的细胞内激活途径触发凋亡。然而,目前对后者机制的了解还很有限。在这里,我们剖析了汉坦病毒感染(以引起 HFRS 的汉坦病毒和引起 HPS 的安第斯病毒为例)导致对 staurosporine 诱导的细胞凋亡产生抗性的机制。与未感染 staurosporine 暴露的细胞相比,感染细胞中观察到较少的活性 caspase-8 和 caspase-9,因此活性 caspase-3 也较少。虽然 staurosporine 暴露的未感染细胞会将大量促凋亡的细胞色素 C 释放到细胞质中,但在感染细胞中未观察到这种情况。此外,汉坦病毒还阻止了 BAX 的激活和线粒体外膜通透性(MOMP)。同时,在汉坦病毒感染的细胞中观察到促生存因子 BCL-2 的水平显著增加。重要的是,通过抑制剂 ABT-737 直接抑制 BCL-2,以及通过 siRNA 沉默 BCL-2,会导致 staurosporine 暴露的汉坦病毒感染细胞发生凋亡。总的来说,我们在这里提供了一个初步的机制,即汉坦病毒通过诱导促生存因子 BCL-2 的表达增加,从而在线粒体水平保护感染细胞免受内在凋亡,从而阻止 MOMPs 及其随后的 caspase 激活,以此来保护感染细胞免受内在凋亡。汉坦病毒用来确保感染细胞存活的多种机制可能有助于其自然宿主的持续感染,并可能在 HFRS 和 HPS 患者的免疫发病机制中发挥作用。