J Virol. 2015 Jan;89(1):312-22. doi

J Virol. 2015 Jan;89(1):312-22. doi: 10.1128/JVI.01572-14. Epub 2014 Oct 15.
Expression of interferon gamma by a recombinant rabies virus strongly attenuates the pathogenicity of the virus via induction of type I interferon.
Barkhouse DA1, Garcia SA1, Bongiorno EK1, Lebrun A1, Faber M2, Hooper DC3.
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Abstract
Previous animal model experiments have shown a correlation between interferon gamma (IFN-γ) expression and both survival from infection with attenuated rabies virus (RABV) and reduction of neurological sequelae. Therefore, we hypothesized that rapid production of murine IFN-γ by the rabies virus itself would induce a more robust antiviral response than would occur naturally in mice. To test this hypothesis, we used reverse engineering to clone the mouse IFN-γ gene into a pathogenic rabies virus backbone, SPBN, to produce the recombinant rabies virus designated SPBNγ. Morbidity and mortality were monitored in mice infected intranasally with SPBNγ or SPBN(-) control virus to determine the degree of attenuation caused by the expression of IFN-γ. Incorporation of IFN-γ into the rabies virus genome highly attenuated the virus. SPBNγ has a 50% lethal dose (LD50) more than 100-fold greater than SPBN(-). In vitro and in vivo mouse experiments show that SPBNγ infection enhances the production of type I interferons. Furthermore, knockout mice lacking the ability to signal through the type I interferon receptor (IFNAR(-/-)) cannot control the SPBNγ infection and rapidly die. These data suggest that IFN-γ production has antiviral effects in rabies, largely due to the induction of type I interferons.
IMPORTANCE:
Survival from rabies is dependent upon the early control of virus replication and spread. Once the virus reaches the central nervous system (CNS), this becomes highly problematic. Studies of CNS immunity to RABV have shown that control of replication begins at the onset of T cell entry and IFN-γ production in the CNS prior to the appearance of virus-neutralizing antibodies. Moreover, antibody-deficient mice are able to control but not clear attenuated RABV from the CNS. We find here that IFN-γ triggers the early production of type I interferons with the expected antiviral effects. We also show that engineering a lethal rabies virus to express IFN-γ directly in the infected tissue reduces rabies virus replication and spread, limiting its pathogenicity in normal and immunocompromised mice. Therefore, vector delivery of IFN-γ to the brain may have the potential to treat individuals who would otherwise succumb to infection with rabies virus.

結果 (中文) 1:

J Virol。2015 Jan; 1:312-22。doi: 10.1128/JVI.01572-14。Epub 2014 Oct 15。通过重组狂犬病病毒干扰素 γ 表达强烈衰减致病性的病毒通过诱导 I 型干扰素。Barkhouse 达 1，加西亚 SA1，开灯会 EK1、 特利尔 A1，费伯 M2，胡珀 DC3。作者信息摘要Previous animal model experiments have shown a correlation between interferon gamma (IFN-γ) expression and both survival from infection with attenuated rabies virus (RABV) and reduction of neurological sequelae. Therefore, we hypothesized that rapid production of murine IFN-γ by the rabies virus itself would induce a more robust antiviral response than would occur naturally in mice. To test this hypothesis, we used reverse engineering to clone the mouse IFN-γ gene into a pathogenic rabies virus backbone, SPBN, to produce the recombinant rabies virus designated SPBNγ. Morbidity and mortality were monitored in mice infected intranasally with SPBNγ or SPBN(-) control virus to determine the degree of attenuation caused by the expression of IFN-γ. Incorporation of IFN-γ into the rabies virus genome highly attenuated the virus. SPBNγ has a 50% lethal dose (LD50) more than 100-fold greater than SPBN(-). In vitro and in vivo mouse experiments show that SPBNγ infection enhances the production of type I interferons. Furthermore, knockout mice lacking the ability to signal through the type I interferon receptor (IFNAR(-/-)) cannot control the SPBNγ infection and rapidly die. These data suggest that IFN-γ production has antiviral effects in rabies, largely due to the induction of type I interferons.IMPORTANCE:Survival from rabies is dependent upon the early control of virus replication and spread. Once the virus reaches the central nervous system (CNS), this becomes highly problematic. Studies of CNS immunity to RABV have shown that control of replication begins at the onset of T cell entry and IFN-γ production in the CNS prior to the appearance of virus-neutralizing antibodies. Moreover, antibody-deficient mice are able to control but not clear attenuated RABV from the CNS. We find here that IFN-γ triggers the early production of type I interferons with the expected antiviral effects. We also show that engineering a lethal rabies virus to express IFN-γ directly in the infected tissue reduces rabies virus replication and spread, limiting its pathogenicity in normal and immunocompromised mice. Therefore, vector delivery of IFN-γ to the brain may have the potential to treat individuals who would otherwise succumb to infection with rabies virus.

結果 (中文) 3:

杂志。2015 Jan；89（1）：312-22。关键词：10.1128/jvi.01572-14。EPUB 2014 Oct 15。
通过重组狂犬病病毒干扰素γ的表达强烈抑制病毒的致病性通过I型干扰素的诱导。
巴克豪斯DA1，加西亚SA1，Bongiorno EK1，勒布伦A1，麦嘉华平方米，Hooper DC3。
作者信息

摘要
以前的动物模型实验表明之间的相关性（IFN-γ干扰素γ）表达与生存率从减毒狂犬病毒感染（病毒）和减少神经系统后遗症。因此，我们推测，快速生产的小鼠干扰素-γ的狂犬病病毒本身会引起一个更强大的抗病毒反应比自然发生在小鼠体内。为了检验这一假设，我们使用逆向工程克隆小鼠IFN-γ基因与致病病毒骨干，SPBN，产生重组狂犬病毒指定SPBNγ。发病率和死亡率进行了监测小鼠滴鼻感染SPBNγ或SPBN（-）控制病毒确定程度的IFN-γ的表达引起的衰减γ。IFN病毒基因组的掺入到狂犬病病毒的高度减毒。γSPBN有50%的致死剂量（LD50）的100倍以上SPBN（-）。在体外和体内小鼠实验表明SPBNγ感染增强I型干扰素的生产。此外，基因敲除小鼠缺乏信号通过I型干扰素受体的能力（IFNAR（-/-））无法控制SPBNγ感染和快速模具。这些数据表明，IFN-γ生产狂犬病抗病毒效果，很大程度上是由于I型干扰素的诱导。
重要性：
生存依赖于从狂犬病病毒的复制和传播的早期控制。一旦病毒到达中枢神经系统（中枢神经系统），这就变得非常麻烦。中枢神经系统免疫狂犬病的研究表明，控制复制的T细胞进入和IFN-γ生产在中枢神经系统中的病毒中和抗体出现之前开始发生。此外，抗体缺陷小鼠能够控制但不清楚减毒狂犬病从中枢神经系统。我们在此发现IFN-γ触发的I型干扰素的抗病毒效果与预期的早期生产。我们也在感染组织减少狂犬病病毒的复制和传播表达IFN-γ直接表明工程致命的狂犬病毒，在正常限制其致病性和免疫功能低下小鼠。因此，干扰素-γ-干扰素的载体传递可能有可能治疗的人，否则会屈服于感染狂犬病病毒。