To Eunice E, Erlich Jonathan R, Liong Felicia, Liong Stella, Luong Raymond, Oseghale Osezua, Miles Mark A, Papagianis Paris C, Quinn Kylie M, Bozinovski Steven, Vlahos Ross, Brooks Robert D, O'Leary John J, Brooks Doug A, Selemidis Stavros
School of Health and Biomedical Sciences, RMIT University, Bundoora, VIC, Australia.
F.M Kirby Neurobiology Centre, Boston Children's Hospital, Harvard Medical School, Boston, MA, United States.
Front Pharmacol. 2022 Mar 23;13:870156. doi: 10.3389/fphar.2022.870156. eCollection 2022.
There is an urgent need to develop effective therapeutic strategies including immunomodulators to combat influenza A virus (IAV) infection. Influenza A viruses increase ROS production, which suppress anti-viral responses and contribute to pathological inflammation and morbidity. Two major cellular sites of ROS production are endosomes the NOX2-oxidase enzyme and the electron transport chain in mitochondria. Here we examined the effect of administration of Cgp91ds-TAT, an endosome-targeted NOX2 oxidase inhibitor, in combination with mitoTEMPO, a mitochondrial ROS scavenger and compared it to monotherapy treatment during an established IAV infection. Mice were infected with IAV (Hkx31 strain; 10PFU/mouse) and 24 h post infection were treated with Cgp91ds-TAT (0.2 mg/kg), mitoTEMPO (100 μg) or with a combination of these inhibitors [Cgp91ds-TAT (0.2 mg/kg)/mitoTEMPO (100 μg)] intranasally every day for up to 2 days post infection (pi). Mice were euthanized on Days 3 or 6 post infection for analyses of disease severity. A combination of Cgp91ds-TAT and mitoTEMPO treatment was more effective than the ROS inhibitors alone at reducing airway and neutrophilic inflammation, bodyweight loss, lung oedema and improved the lung pathology with a reduction in alveolitis following IAV infection. Dual ROS inhibition also caused a significant elevation in Type I IFN expression at the early phase of infection (day 3 pi), however, this response was suppressed at the later phase of infection (day 6 pi). Furthermore, combined treatment with Cgp91ds-TAT and mitoTEMPO resulted in an increase in IAV-specific CD8 T cells in the lungs. In conclusion, this study demonstrates that the reduction of ROS production in two major subcellular sites, i.e. endosomes and mitochondria, by intranasal delivery of a combination of Cgp91ds-TAT and mitoTEMPO, suppresses the severity of influenza infection and highlights a novel immunomodulatory approach for IAV disease management.
迫切需要制定有效的治疗策略，包括使用免疫调节剂来对抗甲型流感病毒（IAV）感染。甲型流感病毒会增加活性氧（ROS）的产生，这会抑制抗病毒反应，并导致病理性炎症和发病。ROS产生的两个主要细胞部位是内体中的NOX2氧化酶和线粒体中的电子传递链。在这里，我们研究了给予Cgp91ds-TAT（一种靶向内体的NOX2氧化酶抑制剂）与线粒体靶向抗氧化剂mitoTEMPO联合使用的效果，并将其与IAV感染确诊后的单一疗法进行比较。小鼠感染IAV（Hkx31株；10PFU/小鼠），感染后24小时开始每天经鼻给予Cgp91ds-TAT（0.2mg/kg）、mitoTEMPO（100μg）或这些抑制剂的组合[Cgp91ds-TAT（0.2mg/kg）/mitoTEMPO（100μg）]，持续至感染后2天（pi）。感染后第3天或第6天对小鼠实施安乐死，以分析疾病严重程度。Cgp91ds-TAT和mitoTEMPO联合治疗在减轻气道和中性粒细胞炎症、体重减轻、肺水肿方面比单独使用ROS抑制剂更有效，并改善了肺病理学，减少了IAV感染后的肺泡炎。双重ROS抑制还在感染早期（感染后第3天）导致I型干扰素表达显著升高，然而，这种反应在感染后期（感染后第6天）受到抑制。此外，Cgp91ds-TAT和mitoTEMPO联合治疗导致肺中IAV特异性CD8 T细胞增加。总之，本研究表明，通过经鼻递送Cgp91ds-TAT和mitoTEMPO的组合来减少两个主要亚细胞部位（即内体和线粒体）中的ROS产生，可抑制流感感染的严重程度，并突出了一种用于IAV疾病管理的新型免疫调节方法。
