Ariza Maria Eugenia, Glaser Ronald, Williams Marshall V
Department of Molecular Virology, Immunology, and Medical Genetics, The Ohio State University College of Medicine Columbus, OH, USA.
Department of Molecular Virology, Immunology, and Medical Genetics, The Ohio State University College of Medicine Columbus, OH, USA ; Institute for Behavioral Medicine Research, The Ohio State University College of Medicine Columbus, OH, USA.
Front Microbiol. 2014 Sep 26;5:504. doi: 10.3389/fmicb.2014.00504. eCollection 2014.
We have previously shown that Epstein-Barr virus (EBV)-encoded dUTPase can modulate innate immune responses through the activation of TLR2 and NF-κB signaling. However, whether this novel immune function of the dUTPase is specific for EBV or a common property of the Herpesviridae family is not known. In this study, we demonstrate that the purified viral dUTPases encoded by herpes simplex virus type 2 (HSV-2), human herpesvirus-6A (HHV-6A), human herpesvirus-8 (HHV-8) and varicella-zoster virus (VZV) differentially activate NF-κB through ligation of TLR2/TLR1 heterodimers. Furthermore, activation of NF-κB by the viral dUTPases was inhibited by anti-TLR2 blocking antibodies (Abs) and the over-expression of dominant-negative constructs of TLR2, lacking the TIR domain, and MyD88 in human embryonic kidney 293 cells expressing TLR2/TLR1. In addition, treatment of human dendritic cells and PBMCs with the herpesviruses-encoded dUTPases from HSV-2, HHV-6A, HHV-8, and VZV resulted in the secretion of the inflammatory cytokines IL-1β, IL-6, IL-8, IL-12, TNF-α, IL-10, and IFN-γ. Interestingly, blocking experiments revealed that the anti-TLR2 Ab significantly reduced the secretion of cytokines by the various herpesviruses-encoded dUTPases (p < 0.05). To our knowledge, this is the first report demonstrating that a non-structural protein encoded by herpesviruses HHV-6A, HHV-8, VZV and to a lesser extent HSV-2 is a pathogen-associated molecular pattern. Our results reveal a novel function of the virus-encoded dUTPases, which may be important to the pathophysiology of diseases caused by these viruses. More importantly, this study demonstrates that the immunomodulatory functions of dUTPases are a common property of the Herpesviridae family and thus, the dUTPase could be a potential target for the development of novel therapeutic agents against infections caused by these herpesviruses.
我们之前已经表明,爱泼斯坦-巴尔病毒(EBV)编码的脱氧尿苷三磷酸酶(dUTPase)可通过激活Toll样受体2(TLR2)和核因子κB(NF-κB)信号传导来调节先天性免疫反应。然而,dUTPase的这种新免疫功能是EBV特有的,还是疱疹病毒科的共同特性,目前尚不清楚。在本研究中,我们证明单纯疱疹病毒2型(HSV-2)、人类疱疹病毒6A型(HHV-6A)、人类疱疹病毒8型(HHV-8)和水痘-带状疱疹病毒(VZV)编码的纯化病毒dUTPase通过TLR2/TLR1异二聚体的连接差异激活NF-κB。此外,在表达TLR2/TLR1的人胚肾293细胞中,病毒dUTPase对NF-κB的激活被抗TLR2阻断抗体(Abs)以及缺乏TIR结构域的TLR2显性负性构建体和髓样分化因子88(MyD88)的过表达所抑制。此外,用HSV-2、HHV-6A、HHV-8和VZV的疱疹病毒编码的dUTPase处理人树突状细胞和外周血单个核细胞(PBMCs)会导致炎性细胞因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-12(IL-12)、肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)和干扰素-γ(IFN-γ)的分泌。有趣的是,阻断实验表明,抗TLR2抗体显著降低了各种疱疹病毒编码的dUTPase分泌的细胞因子(p<0.05)。据我们所知,这是第一份证明HHV-6A、HHV-8、VZV以及程度较轻的HSV-2编码的非结构蛋白是一种病原体相关分子模式的报告。我们的结果揭示了病毒编码dUTPase的一种新功能,这可能对这些病毒引起的疾病的病理生理学很重要。更重要的是,本研究表明dUTPase的免疫调节功能是疱疹病毒科的共同特性,因此,dUTPase可能是开发针对这些疱疹病毒感染的新型治疗药物的潜在靶点。