The Induction of Pattern-Recognition Receptor Expression against Influenza A Virus through Duox2-Derived Reactive Oxygen Species in Nasal Mucosa

Abstract

We studied the relative roles of Duox2-derived reactive oxygen species (ROS) in host defense against influenza A virus (IAV) infection in normal human nasal epithelial cells and mouse nasal mucosa. We found that Duox2 primarily generated ROS rapidly after IAV infection in normal human nasal epithelial cells and that knockdown of Duox2 aggravated IAV infection. In addition, Duox2-derived ROS enhancement significantly suppressed IAV infection in nasal epithelium. In particular, Duox2-derived ROS were required for the induction of retinoic acid-inducible gene (RIG)-I and melanoma differentiation-associated protein 5 (MDA5) transcription. After intranasal IAV inoculation into mice, viral infection was significantly aggravated from 3 days postinoculation (dpi) in the nasal mucosa, and the IAV viral titer was highest at 7 dpi. Both RIG-I and MDA5 messenger RNA levels increased dominantly in mouse nasal mucosa from 3 dpi; consistent with this, RIG-I and MDA5 proteins were also induced after IAV infection. RIG-I and MDA5 messenger RNA levels were induced to a lower extent in the nasal mucosa of the mice that were inoculated with Duox2 short hairpin RNA, and the IAV viral titer was significantly higher in nasal lavage. Taken together, Duox2-derived ROS are necessary for the innate immune response and trigger the induction of RIG-I and MDA5 to resist IAV infection in human nasal epithelium and mouse nasal mucosa.

Chemiluminescence imaging of Duox2-derived hydrogen peroxide for longitudinal visualization of biological response to viral infection in nasal mucosa

“…Among isoforms of NADPH oxidase, dual oxidase 2 (Duox2) has been shown to be a major one involved in the H 2 O 2 generation in the respiratory tract 1 - 3 , 18 . Interestingly, a recent work has described Duox2-generated H 2 O 2 as an integral part of the host defense system at mucosal surfaces 11 , 12 , 18 , 19 , which was verified by the demonstration that a lack of Duox2-derived H 2 O 2 in respiratory epithelial cells causes fatal viral infection in animal disease models 13 . According to this view, there has been growing interest in longitudinal evaluation of Duox2 activity in order to gain a better insight into the innate immunity in the respiratory tract.…”

Mini review: immunologic functions of dual oxidases in mucosal systems of vertebrates

“…In a perinatal lamb model, after RSV infection, DUOX expression levels were increased under bronchiolitis (Derscheid and Ackermann, 2012). Retinoic acid inducible gene 1 and melanoma differentiation-associated protein 5 also can induce DUOX2-mediated antiviral responses (Kim et al, 2015). In chronic rhinosinusitis (CRS), DUOX mRNA levels were significantly elevated.…”

Innate Immunity of the Lung: From Basic Mechanisms to Translational Medicine

“…These studies using nasal biopsies and primary human nasal epithelial cells stimulated with artificial TLR and RLR agonists demonstrated how these receptors generate robust IL-6, IL-8, and interferon (IFN)-β responses and act as a first line of defense against viruses invading the respiratory tract. In another study, reactive oxygen species produced via Duox2 in influenza A virus-infected nasal epithelial cells were shown to increase RIG-I and MDA-5 mRNA in a process believed to resist IAV infection [13]. Kim et al [14] also evaluated inflammatory and antiviral responses to human virus infection in human nasal epithelial cells from chronic rhinosinusitis patients, wherein decreased MDA-5 and IFN-β expressions were evident and human rhinovirus clearance was slightly impaired.…”