Manipulating adenovirus hexon hypervariable loops dictates immune neutralisation and coagulation factor X-dependent cell interaction in vitro and in vivo.
Manipulating adenovirus hexon hypervariable loops dictates immune neutralisation and coagulation factor X-dependent cell interaction in vitro and in vivo.
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DOI：
10.1371/journal.ppat.1004673
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发表时间：
2015-02
期刊：
影响因子：
6.7
通讯作者：
Baker AH
中科院分区：
文献类型：
--
作者：
Ma J;Duffy MR;Deng L;Dakin RS;Uil T;Custers J;Kelly SM;McVey JH;Nicklin SA;Baker AH
腺病毒是常见的病原体，主要针对眼、胃肠道和呼吸道细胞，但在某些情况下感染会传播，导致严重的临床结果。在传播和与血液接触后，凝血因子X （FX）直接与腺病毒5型（Ad5）六邻体相互作用。FX可以作为结合硫酸肝素蛋白聚糖的桥梁，导致Ad5肝细胞大量摄取。FX“涂层”也保护病毒免受宿主IgM和补体介导的中和。然而，FX在确定Ad -肝转导同时屏蔽病毒免受免疫攻击方面的作用尚不清楚。在这项研究中，我们证明了FX保护机制在Ad类型中并不保守，并确定了Ad5的六邻体高变区（HVR）是这种宿主防御途径的衣壳蛋白。使用遗传和药理学方法，我们操纵Ad5 HVR相互作用来询问病毒细胞转导和免疫中和之间的相互作用。我们发现FX和抑制性血清成分可以共同竞争，病毒中和受到Ad5 hvr修饰的位置和程度的影响。我们将ad5衍生的hvr植入罕见的原生非fx结合Ad26中，从而创建Ad26. hvr5c。通过表面等离子体共振、FX介导的细胞结合和转导试验，这使得病毒能够以高亲和力与FX相互作用。与此同时,Ad26。HVR5C在缺乏FX的情况下也对免疫攻击敏感，这是来自Ad5的工程化HVRs的直接后果。在免疫能力强和免疫缺陷的动物中，Ad26。静脉给药后FX介导HVR5C肝基因转移。这项研究为Ad5 HVRs在IgM和经典补体介导的攻击中赋予病毒中和敏感性的关键作用提供了机制见解。此外，通过这种功能增益方法，我们展示了FX在保护Ad26方面的双重功能。HVR5C对抗先天免疫因子，同时确定肝脏靶向性。腺病毒通常被认为是与呼吸道、胃肠道和眼部感染相关的自限性病原体；然而，在免疫功能低下的受试者中，播散性Ad感染可发生危及生命的后果。许多人的ad能够与凝血因子X （FX）结合。在动物模型中静脉给药后，FX直接与主要的Ad衣壳蛋白六元结合，随后导致病毒在肝脏中积聚。FX涂层Ad5还可以通过天然IgM抗体和经典补体系统屏蔽免疫中和。在这里，我们证明FX保护在ad中不是一个保守的机制，并确定Ad5六元高变区（HVR）是这种宿主防御途径的衣壳蛋白靶向。此外，研究人员发现，将Ad5 hvr基因包被在原生非fx结合物Ad26上，足以在体外和体内赋予对免疫攻击的敏感性。通过静脉给药，我们确定FX与ad5衍生的hvr结合在保护病毒免受中和的同时介导病毒趋向性方面的重要性。我们的研究为病毒HVRs和FX在病毒和宿主防御机制之间的界面中的作用提供了新的见解。
Adenoviruses are common pathogens, mostly targeting ocular, gastrointestinal and respiratory cells, but in some cases infection disseminates, presenting in severe clinical outcomes. Upon dissemination and contact with blood, coagulation factor X (FX) interacts directly with the adenovirus type 5 (Ad5) hexon. FX can act as a bridge to bind heparan sulphate proteoglycans, leading to substantial Ad5 hepatocyte uptake. FX “coating” also protects the virus from host IgM and complement-mediated neutralisation. However, the contribution of FX in determining Ad liver transduction whilst simultaneously shielding the virus from immune attack remains unclear. In this study, we demonstrate that the FX protection mechanism is not conserved amongst Ad types, and identify the hexon hypervariable regions (HVR) of Ad5 as the capsid proteins targeted by this host defense pathway. Using genetic and pharmacological approaches, we manipulate Ad5 HVR interactions to interrogate the interplay between viral cell transduction and immune neutralisation. We show that FX and inhibitory serum components can co-compete and virus neutralisation is influenced by both the location and extent of modifications to the Ad5 HVRs. We engineered Ad5-derived HVRs into the rare, native non FX-binding Ad26 to create Ad26.HVR5C. This enabled the virus to interact with FX at high affinity, as quantified by surface plasmon resonance, FX-mediated cell binding and transduction assays. Concomitantly, Ad26.HVR5C was also sensitised to immune attack in the absence of FX, a direct consequence of the engineered HVRs from Ad5. In both immune competent and deficient animals, Ad26.HVR5C hepatic gene transfer was mediated by FX following intravenous delivery. This study gives mechanistic insight into the pivotal role of the Ad5 HVRs in conferring sensitivity to virus neutralisation by IgM and classical complement-mediated attack. Furthermore, through this gain-of-function approach we demonstrate the dual functionality of FX in protecting Ad26.HVR5C against innate immune factors whilst determining liver targeting. Adenoviruses are mostly considered self-limiting pathogens associated with respiratory, gastrointestinal and ocular infections; however, in immunocompromised subjects disseminated Ad infection can occur with life-threatening consequences. Many human Ads are capable of binding to coagulation factor X (FX). Following intravenous administration in animal models, FX binds directly to the major Ad capsid protein, the hexon, which subsequently results in virus accumulation in the liver. FX coating Ad5 also acts to shield against immune neutralisation via natural IgM antibodies and the classical complement system. Here we show that FX protection is not a conserved mechanism amongst Ads and identify the Ad5 hexon hypervariable regions (HVR) as the capsid proteins targeted by this host defense pathway. Furthermore, we show that genetic inclusion of Ad5 HVRs onto a native non-FX binder Ad26 to be sufficient to confer sensitivity to immune attack in vitro and in vivo. Using intravenous administration, we determine the significance of FX binding to the Ad5-derived HVRs with respect to defending the virus from neutralisation whilst mediating virus tropism. Our study gives new insight into the role of the viral HVRs and of FX at the interface between virus and host defense mechanisms.
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影响因子：
5.4
作者：
Irons, Eric E.;Flatt, Justin W.;Shayakhmetov, Dmitry M.
通讯作者：
Shayakhmetov, Dmitry M.
影响因子：
5.4
作者：
Chen, H.;Xiang, Z. Q.;Ertl, H. C. J.
通讯作者：
Ertl, H. C. J.
影响因子：
5.4
作者：
Duffy, Margaret R.;Bradshaw, Angela C.;Baker, Andrew H.
通讯作者：
Baker, Andrew H.
DOI：
10.1126/science.1226625
发表时间：
2012-11-09
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Doronin K;Flatt JW;Di Paolo NC;Khare R;Kalyuzhniy O;Acchione M;Sumida JP;Ohto U;Shimizu T;Akashi-Takamura S;Miyake K;MacDonald JW;Bammler TK;Beyer RP;Farin FM;Stewart PL;Shayakhmetov DM
通讯作者：
Shayakhmetov DM
影响因子：
4.8
作者：
Chakrabarti, S;Collingham, KE;Milligan, DW
通讯作者：
Milligan, DW