Elsevier

Brain, Behavior, and Immunity

Volume 88, August 2020, Pages 353-362
Brain, Behavior, and Immunity

Full-length Article
S100A9 plays a pivotal role in a mouse model of herpetic neuralgia via TLR4/TNF pathway

https://doi.org/10.1016/j.bbi.2020.03.033Get rights and content

Highlights

  • S100A9 is overexpressed in DRGs after HSV-1 infection.

  • S100A9 positively modulate herpetic neuralgia.

  • Neutrophils are the main source of S100A9 in DRGs after HSV-1 infection.

  • S100A9 mediates herpetic neuralgia via the TLR4/TNF pathway.

Abstract

Herpetic neuralgia is a painful condition following herpes zoster disease, which results from Varicella-zoster virus reactivation in the dorsal or trigeminal sensory ganglia. Nevertheless, the pathophysiological mechanisms involved in herpetic neuralgia are not well understood. Recently, we identified, that neuroimmune-glia interactions in the sensory ganglion is a critical mechanism for the development of herpetic neuralgia. Here, we investigate the contribution of S100A9, a well-known pro-inflammatory molecule produced by myeloid cells, for the development of herpetic neuralgia using a murine model of HSV-1 infection. We found that cutaneous HSV-1 infection results in an increase of S100A9 expression in the Dorsal Root Ganglia (DRGs). Infiltrating neutrophils into the DRGs were the main source of S100A9 post HSV-1 infection. Functionally, genetic or pharmacological inhibition of S100A9 impairs the development of HSV-1 infection-induced mechanical pain hypersensitivity. Finally, we found that the pronociceptive role of S100A9 in herpetic neuralgia depends on the TLR4/TNF pathway. These results unraveled previously unknown mechanisms involved in the pathophysiology of herpetic neuralgia and indicate that S100A9 might be an important target for novel therapies aiming acute herpetic neuralgia.

Introduction

Herpetic neuralgia is the most important symptom of herpes zoster disease, which is caused by Varicella zoster Virus (VZV) reactivation in the dorsal root (DRGs) or trigeminal ganglia post primary infection (Nagel and Gilden, 2014, Opstelten et al., 2010). Herpes zoster affects approximately 25% of the population and the resulting neuralgia is clinically characterized by a painful blistering skin rash which negatively impacts the quality of life of the individuals and the disease management is challenging and often unsatisfactory (Johnson, 2010, Opstelten et al., 2010). Mice are resistant to VZV infection, however herpes simplex virus type-1 (HSV-1) has been employed to infect animals. HSV-1 is a neurotropic virus that infect the neurons and induces a zosteriform-like skin lesion and herpetic pain, allowing important advances in the understanding of the pathophysiological mechanisms involved in the development of herpes zoster and herpetic neuralgia (Silva et al., 2017, Takasaki et al., 2000a, Takasaki et al., 2000b, Wilson and Mohr, 2012).

Recently, we have shown that neuroimmune-glia interactions in sensory ganglia play a critical role in the development of herpetic neuralgia (Silva et al., 2017). For instance, we have shown that infiltrating leukocytes, mostly neutrophils and macrophages, reach the infected DRGs and contributes to the development of herpetic neuralgia. These leukocytes are mediating herpetic neuralgia mainly through the production of inflammatory mediators such as tumor necrosis factor (TNF) (Silva et al., 2017). Infiltrating myeloid cells, including neutrophils and activated macrophages, can express and secrete a cytoplasmic pro-inflammatory protein, S100 calcium-binding protein A9 (S100A9), which presents a broad range of pro-inflammatory activities including the promotion of immune cells activation and proinflammatory cytokine secretion (Chen et al., 2014, Edgeworth et al., 1991, Mitchell et al., 2008, Tsai et al., 2014, Tsai et al., 2014, Xu et al., 2013). S100A9, also known as myeloid related protein 14 (MRP14), belong to a S100 protein subfamily termed myeloid-related proteins, which are linked to innate immune functions which can trigger the production of reactive oxygen species (ROS) and inflammatory cytokines including TNFα, IL1β, IL6, and IL8 (Cheng et al., 2008, De Veirman et al., 2017, Sunahori et al., 2006). The use of S100A9-deficient mice revealed a pivotal role of this protein in the pathophysiology of several inflammatory diseases such as arthritis, sepsis, multiple sclerosis, inflammatory bowel disease, cancers and neurodegenerative disorders (Holzinger et al., 2014, Kovačić et al., 2018, Lügering et al., 1995, Roth et al., 1992, Schonthaler et al., 2013). Regarding the molecular mechanisms by which S100A9 triggers innate immune response, there are evidences showing it could activate Toll-like receptor 4 (TLR4) resulting in cytokine production and increased inflammation (Holzinger et al., 2014, Nishikawa et al., 2017, Tsai et al., 2014, Zheng et al., 2014).

However, despite the well-known role of S100A9 in the pathophysiology of inflammatory and infection diseases, its participation in herpetic neuralgia has been never investigated. Thus, we aimed to investigate the role of S100A9 and its signaling in an animal model of herpetic neuralgia induced by HSV-1 infection in mice.

Section snippets

Animals

The experiments were performed on male C57BL/6 wild type (WT), S100a9 and Tlr4 deficient (−/−) mice (C57BL/6 background). Their colonies were maintained in the animal facility of Ribeirão Preto Medical School, University of São Paulo, Brazil and used at weight of 20–25 g. They were housed in temperature-controlled rooms (22–25 °C) and given water and food ad libitum at the animal facility at the Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Brazil. Animal

HSV-1 peripheral infection is able to induce acute herpetic neuralgia in mice

Firstly, we analyzed the ability of HSV-1 skin infection to induce skin lesion and mechanical pain hypersensitivity in mice, important characteristics of herpetic neuralgia, using a murine model (Fig. 1A). HSV-1 skin infection induces the development of zosteriform-like lesions in around 60% (n 6/10) of mice, which can be visualized from 3 days post-infection, progressing until 7 dpi (Fig. 1B) and disappearing at 21 dpi, as demonstrated by Silva 2017 (Silva et al., 2017). No zosteriform-like

Discussion

Acute herpes zoster is clinically characterized by the presence of a zosteriform skin lesion accompanied by spontaneous and evoked severe pain as tactile allodynia, starting after the reactivation of a dormant varicella-zoster virus in the sensory ganglia (Loeser, 1986, Opstelten et al., 2010). We have previously shown that neuro-immune interactions at the level of sensory ganglia play a fundamental role in the development of HSV-1 induced acute herpetic neuralgia (Silva et al., 2017). Here, we

Acknowledgments

The authors gratefully acknowledge the technical assistance of Sergio R. Rosa, Ieda Santos, Katia Santos, and Sonia Aparecida Zanon. We also would like to acknowledge to the Active Biotech AB, which provided the paquinimod compound. The research leading to these results has received funding from São Paulo Research Foundation (FAPESP) under grant agreements n°2011/19670-0 (Thematic project) and 2013/08216-2 (Center for Research in Inflammatory Disease) and Brazilian National Council for

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