Franca Rafael F O, Costa Renata S, Silva Jaqueline R, Peres Raphael S, Mendonça Leila R, Colón David F, Alves-Filho José Carlos, Cunha Fernando Q
Department of Virology and Experimental Therapy LAVITE, Oswaldo Cruz Foundation - FIOCRUZ, Institute Aggeu Magalhães IAM, Av. Professor Moraes Rego, s/n, Recife, PE, 50740-465, Brazil.
Program of Basic and Applied Immunology, University of São Paulo, Ribeirao Preto, SP, 14049-900, Brazil.
J Neuroinflammation. 2016 Jun 22;13(1):159. doi: 10.1186/s12974-016-0628-1.
Viral encephalitis is a common cause of lethal infections in humans, and several different viruses are documented to be responsible. Rocio virus is a flavivirus that causes a severe lethal encephalitis syndrome in humans and also mice, providing an interesting model to study the CNS compartmentalized immune response. Interleukin 33 (IL-33), a member of the IL-1 family, is an immunomodulatory cytokine that is highly expressed in the CNS. However, the role of IL-33 on viral encephalitis remains unclear. Therefore, we aimed to explore how the IL-33/ST2 axis regulates the local immune response during Rocio virus infection.
Wild-type (WT), ST2 (ST2(-/-)), and nitric oxide synthase-deficient mice (iNOS(-/-)) and Stat6 (Stat6(-/-))-deficient mice were infected with different concentrations of the Rocio virus by intraperitoneal route, the cytokine mRNA level in CNS was analyzed by qPCR, and cellular immunophenotyping was performed on infected mice by the flow cytometry of isolated CNS mononuclear cells.
We have shown that the mRNA expression of IL-33 and ST2 receptors is increased in the CNS of Rocio virus-infected WT mice and that ST2(-/-) mice showed increased susceptibility to infection. ST2 deficiency was correlated with increased tissue pathology, cellular infiltration, and tumor necrosis factor alpha (TNF-α) and interferon-gamma (IFN-γ) mRNA levels and higher viral load in the CNS, compared with wild-type mice. The increased Th1 cytokine levels released in the CNS acted on infiltrating macrophages, as evidenced by flow cytometry characterization of cellular infiltrates, inducing the expression of iNOS, contributing to brain injury. Moreover, iNOS(-/-) mice were more resistant to Rocio virus encephalitis, presenting a lower clinical score and reduced mortality rate, despite the increased tissue pathology.
We provide evidences of a specific role for IL-33 receptor signaling in nitric oxide induction through local IFN-γ modulation, suggesting that nitric oxide overproduction might have an important role in the progression of experimental viral encephalitis.
病毒性脑炎是人类致死性感染的常见病因,已证实多种不同病毒与之相关。罗西奥病毒是一种黄病毒,可在人类和小鼠中引发严重的致死性脑炎综合征,为研究中枢神经系统(CNS)的局部免疫反应提供了一个有趣的模型。白细胞介素33(IL-33)是IL-1家族的一员,是一种在中枢神经系统中高表达的免疫调节细胞因子。然而,IL-33在病毒性脑炎中的作用仍不清楚。因此,我们旨在探讨IL-33/ST2轴在罗西奥病毒感染期间如何调节局部免疫反应。
野生型(WT)、ST2(ST2(-/-))、一氧化氮合酶缺陷小鼠(iNOS(-/-))和信号转导和转录激活因子6(Stat6(-/-))缺陷小鼠通过腹腔途径感染不同浓度的罗西奥病毒,采用定量聚合酶链反应(qPCR)分析中枢神经系统中的细胞因子mRNA水平,并通过对分离的中枢神经系统单核细胞进行流式细胞术对感染小鼠进行细胞免疫表型分析。
我们发现,在感染罗西奥病毒的野生型小鼠的中枢神经系统中,IL-33和ST2受体的mRNA表达增加,并且ST2(-/-)小鼠对感染的易感性增加。与野生型小鼠相比,ST2缺陷与组织病理学加重、细胞浸润、肿瘤坏死因子α(TNF-α)和干扰素-γ(IFN-γ)mRNA水平升高以及中枢神经系统中病毒载量增加相关。中枢神经系统中释放的Th1细胞因子水平升高作用于浸润的巨噬细胞,这通过细胞浸润的流式细胞术表征得到证实,诱导诱导型一氧化氮合酶(iNOS)的表达,导致脑损伤。此外,尽管组织病理学加重,iNOS(-/-)小鼠对罗西奥病毒脑炎更具抵抗力,表现出较低的临床评分和死亡率。
我们提供了证据表明IL-33受体信号通过局部IFN-γ调节在一氧化氮诱导中具有特定作用,提示一氧化氮的过度产生可能在实验性病毒性脑炎的进展中起重要作用。