Virology最新文献_第9页

RNAi therapeutics targeting Japanese encephalitis virus: Gene targets, delivery platforms, and translational barriers 针对日本脑炎病毒的RNAi疗法：基因靶标、传递平台和翻译障碍

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-10 DOI: 10.1016/j.virol.2025.110713

Maneesh Kumar , Pratima Gupta , Suman Kumar , Shyam Kishor Kumar , Shashank Nand Tiwari , Ashok Aspatwar

Japanese encephalitis virus (JEV), a neurotropic flavivirus, is a leading cause of viral encephalitis in Asia, particularly affecting children and causing significant morbidity and mortality. Despite the availability of vaccines and vector control strategies, their limited efficacy against emerging genotypes and incomplete coverage necessitate alternative antiviral approaches. RNA interference (RNAi) has emerged as a promising therapeutic modality by exploiting post-transcriptional gene silencing to inhibit viral replication. Notably, conserved viral genes—such as capsid (C), envelope (E), NS3, and NS5—represent optimal RNAi targets across JEV genotypes due to their essential roles in the viral life cycle and minimal sequence variability. Recent advances in delivery platforms, including lipid nanoparticles, lentiviral vectors, and artificial microRNAs, have significantly improved RNAi stability, blood-brain barrier (BBB) penetration, and cell-specific targeting. Comparative insights from related flaviviruses such as Dengue and Zika have further informed effective RNAi design. Preclinical studies have demonstrated potent antiviral effects, with >90 % suppression of viral RNA, substantial reductions in brain viral load, and enhanced survival in murine models. However, clinical translation remains challenged by delivery efficiency, immune activation, off-target effects, and the potential for viral escape mutations. This review summarizes the current landscape of RNAi-based therapeutics for JEV, emphasizing conserved gene targets, delivery innovations, and translational hurdles, and outlines future directions for integrating RNAi into next-generation antiviral strategies.

日本脑炎病毒（JEV）是一种嗜神经黄病毒，是亚洲病毒性脑炎的主要病因，尤其影响儿童，并导致大量发病率和死亡率。尽管已有疫苗和病媒控制战略，但它们对新出现的基因型的效力有限，而且覆盖范围不全，因此需要采用其他抗病毒方法。RNA干扰（RNAi）已成为一种有前途的治疗方式，利用转录后基因沉默来抑制病毒复制。值得注意的是，保守的病毒基因，如衣壳(C)、包膜(E)、NS3和ns5，由于它们在病毒生命周期中的重要作用和最小的序列变异性，在所有JEV基因型中都是最佳的RNAi靶点。包括脂质纳米颗粒、慢病毒载体和人工microrna在内的递送平台的最新进展，显著提高了RNAi的稳定性、血脑屏障（BBB）渗透和细胞特异性靶向。来自相关黄病毒（如登革热和寨卡病毒）的比较见解进一步为有效的RNAi设计提供了信息。临床前研究已经证明了有效的抗病毒作用，在小鼠模型中抑制了90%的病毒RNA，大幅降低了脑病毒载量，提高了生存率。然而，临床翻译仍然受到递送效率、免疫激活、脱靶效应和病毒逃逸突变潜力的挑战。本文综述了基于RNAi的乙脑病毒治疗方法的现状，强调了保守的基因靶点、递送创新和翻译障碍，并概述了将RNAi整合到下一代抗病毒策略中的未来方向。

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引用次数: 0

Impact of baseline humoral immunity on treatment outcomes with molnupiravir in the MOVe-OUT randomized, controlled trial 在MOVe-OUT随机对照试验中，基线体液免疫对molnupiravir治疗结果的影响。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-09 DOI: 10.1016/j.virol.2025.110710

Matthew G. Johnson, Julie M. Strizki, David W. Hilbert, Ying Zhang, Patricia Carmelitano, Michelle L. Brown, Dominik J. Wolf, Amanda Paschke, Carisa S. De Anda

Background

In the large, randomized, placebo-controlled MOVe-OUT trial, molnupiravir reduced hospitalization/death in unvaccinated, high-risk adults with mild-to-moderate COVID-19. We investigated humoral immune responses and the impact of anti-SARS-CoV-2 antibodies in MOVe-OUT.

Methods

All participants prospectively underwent serum testing for total antibodies (combined IgA, IgG, and/or IgM) to the SARS-CoV-2 nucleocapsid protein and neutralizing antibodies to the spike protein. For participants with a positive baseline anti-nucleocapsid test, samples were retrospectively tested for subtype-specific IgM and IgG anti-nucleocapsid antibodies. Residual baseline samples from all participants were also retrospectively tested for anti-spike IgG antibodies. Clinical and virologic outcomes according to baseline antibody status were determined.

Results

Fewer participants in the molnupiravir (239/709, 33.7 %) than the placebo arm (259/699, 37.1 %) were positive for any baseline anti–SARS-CoV-2 antibody; anti-spike antibodies only, the specific pattern potentially indicating prior vaccination, were detected in 13.8 % molnupiravir-treated and 15.2 % placebo participants. Molnupiravir reduced the risk of hospitalization/death through day 29 in participants who were seronegative at baseline (molnupiravir 40/470 [8.5 %], placebo 64/440 [14.5 %], 95 % confidence interval for the risk difference: −10.3 % to −1.9 %). Post-baseline, most participants exhibited anti–SARS-CoV-2 antibodies by day 10; by day 29, anti-nucleocapsid antibodies were detected in 93.2 % of molnupiravir-treated and 94.9 % of placebo participants and neutralizing anti-SARS-CoV-2 antibodies in 80.8 % and 82.8 %, respectively.

Conclusion

Anti–SARS-CoV-2 antibodies at baseline, more common with placebo, provided protection against poor clinical outcomes. Potentially undisclosed vaccination unlikely impacted the overall trial outcomes. Molnupiravir treatment did not significantly affect development of humoral immunity to SARS-CoV-2 infection. ClinicalTrials.gov: NCT04575597.

背景：在一项大型、随机、安慰剂对照的MOVe-OUT试验中，molnupiravir降低了未接种疫苗、患有轻中度COVID-19的高危成人的住院/死亡。我们在MOVe-OUT中研究了体液免疫反应和抗sars - cov -2抗体的影响。方法：所有参与者前瞻性地进行了SARS-CoV-2核衣壳蛋白总抗体（IgA、IgG和/或IgM联合抗体）和刺突蛋白中和抗体的血清检测。对于基线抗核衣壳试验阳性的参与者，回顾性检测样品的亚型特异性IgM和IgG抗核衣壳抗体。所有参与者的剩余基线样本也进行了抗刺突IgG抗体的回顾性检测。根据基线抗体状态确定临床和病毒学结果。结果：monupiravir组（239/709,33.7%）比安慰剂组（259/699,37.1%）的任何基线抗sars - cov -2抗体阳性的参与者较少；仅抗刺突抗体（可能表明先前接种过疫苗的特定模式）在13.8%的molnupirvir治疗组和15.2%的安慰剂组中被检测到。在基线时血清阴性的受试者中，Molnupiravir降低了住院/死亡风险至第29天（Molnupiravir 40/470[8.5%]，安慰剂64/440[14.5%]，95%可信区间的风险差异：- 10.3%至- 1.9%）。基线后，大多数参与者在第10天表现出抗sars - cov -2抗体；到第29天，接受molnupirvir治疗的93.2%和安慰剂的94.9%的参与者检测到抗核衣壳抗体，分别在80.8%和82.8%的参与者中检测到抗sars - cov -2抗体。结论：抗sars - cov -2抗体在基线时（在安慰剂组更常见）提供了对不良临床结果的保护。可能未公开的疫苗接种不太可能影响总体试验结果。莫努匹拉韦治疗对SARS-CoV-2感染体液免疫的发展无显著影响。Clinicaltrials: gov: NCT04575597。

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引用次数: 0

Decoding extracellular vesicles-mediated encapsulation of enveloped and nonenveloped gut viruses through phosphatidylserine affinity profiling 通过磷脂酰丝氨酸亲和分析解码细胞外囊泡介导的包膜和非包膜肠道病毒的包封。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-08 DOI: 10.1016/j.virol.2025.110712

Alex Ujong Obeten , Ricardo David Avellán-Llaguno , Haining Huang , Yu-Han Yin , Yue Zhu , Xue-Li Xu , Jing-Yu Chen , Yanzi Wang , GuoZhu Ye , Zhizhen Pan , Li-Ting Zhu , Qiansheng Huang

Viruses are generally classified as enveloped viruses (EnVs) or nonenveloped viruses (non-EnVs), based on the presence of a lipid membrane, with membrane-mediated transmission traditionally attributed to EnVs. However, the composition and characteristics of viral populations encapsulated within extracellular vesicles (EVs) which are phospholipid bilayer nanoparticles released by all living organisms remain poorly understood. Here, we applied a phosphatidylserine (PS)-affinity enrichment strategy to isolate EV-encapsulated viral populations from human stool-derived extracellular viral-like particles (VLPs). Quantitative particle analysis revealed that EnVs exhibited an 11-fold higher PS affinity compared to free non-EnVs (fold change 2.79 vs 0.25). Metagenomic analysis revealed significant enrichment of non-EnVs within PS-positive fractions, including DNA viruses Salasmaviridae (3.84 ± 6.44 %) and RNA bacteriophage Fiersviridae (44.99 ± 32.80 %). Predicted Host-virus correlation analysis highlighted strong correlations between viral families Autographiviridae, Microviridae and host family Enterobacteriaceae. Functional annotation further showed enrichment of structural and replication-related genes in the EV-associated virome. These findings provide evidence for EVs-mediated encapsulation of non-EnVs, challenging the traditional dichotomy of viral classification. This noteworthy observation positions EVs encapsulation as a critical determinant in viral life cycles and underscores the need to revisit current viral taxonomy systems.

基于脂质膜的存在，病毒通常分为包膜病毒（env）和非包膜病毒（non- env），膜介导传播传统上归因于包膜病毒。然而，包裹在细胞外囊泡（EVs）内的病毒群的组成和特征仍然知之甚少，EVs是所有生物体释放的磷脂双层纳米颗粒。在这里，我们应用磷脂酰丝氨酸（PS）亲和富集策略从人粪便来源的细胞外病毒样颗粒（VLPs）中分离出ev封装的病毒群。定量粒子分析显示，与自由的非env相比，env的PS亲和力高11倍（倍差为2.79比0.25）。宏基因组分析显示，非env在ps阳性组分中显著富集，包括DNA病毒Salasmaviridae（3.84±6.44%）和RNA噬菌体Fiersviridae（44.99±32.80%）。预测的宿主-病毒相关性分析强调了病毒科自噬病毒科、微病毒科和宿主科肠杆菌科之间的强相关性。功能注释进一步表明，在ev相关病毒体中存在结构和复制相关基因的富集。这些发现为ev介导的非env包封提供了证据，挑战了传统的病毒分类二分法。这一值得注意的观察表明，ev的封装是病毒生命周期的关键决定因素，并强调需要重新审视当前的病毒分类系统。

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引用次数: 0

Inhibitory effects of berberine hydrochloride on porcine epidemic diarrhea virus in vitro and in vivo 盐酸小檗碱对猪流行性腹泻病毒的体内体外抑制作用。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-07 DOI: 10.1016/j.virol.2025.110711

Mengkai Cai , Zhi Wei , Zhu Wu , Zaizhong Zhang , Weilin Xu , Guihong Zhang , Zezhong Zheng , Lang Gong , Meidi Li

Porcine epidemic diarrhea virus (PEDV) causes severe economic losses in swine, necessitating the development of novel antiviral strategies. In this study, we evaluated the inhibitory effects of berberine hydrochloride (BBH) on PEDV in vitro and in vivo. PEDV-infected Vero-E6 cells were treated with BBH, followed by assays for cell viability, viral RNA levels, and protein expression. PEDV-infected piglets were administered BBH, and clinical signs, intestinal morphology, and viral load were assessed. In vitro, BBH (≤120 μM) exhibited low cytotoxicity and inhibited PEDV replication in a dose-dependent manner, markedly reducing viral titers and nucleocapsid (N) protein expression. Time-of-addition assays showed inhibition across multiple stages of the viral life cycle, with pronounced effects on viral attachment and internalization. Mechanistically, BBH suppressed the autophagy marker LC3-II in a dose- and time-dependent manner, and its antiviral activity was partially reversed by the autophagy activator rapamycin, indicating that autophagy suppression contributes to its effect. In PEDV-challenged suckling piglets, oral BBH (40 mg/kg/day) notably improved survival (66.7 % vs. 0 % in controls), alleviated diarrhea, reduced viral shedding in anal swabs, lowered viral load in intestinal tissues (duodenum, jejunum, ileum), and mitigated histopathological damage. Collectively, these results demonstrate that BBH exerts potent anti-PEDV activity through inhibition of viral entry/replication and modulation of autophagy modulation, supporting its potential as an adjunctive therapy against PEDV.

猪流行性腹泻病毒（PEDV）给猪造成了严重的经济损失，需要开发新的抗病毒策略。在本研究中，我们评估了盐酸小檗碱（BBH）对PEDV的体外和体内抑制作用。用BBH处理pedv感染的Vero-E6细胞，然后检测细胞活力、病毒RNA水平和蛋白质表达。对感染pedv的仔猪进行BBH，并对其临床症状、肠道形态和病毒载量进行评估。在体外，BBH（≤120 μM）表现出较低的细胞毒性，并以剂量依赖的方式抑制PEDV的复制，显著降低病毒滴度和核衣壳(N)蛋白的表达。添加时间测试显示，在病毒生命周期的多个阶段抑制，对病毒的附着和内化有明显的影响。在机制上，BBH以剂量和时间依赖的方式抑制自噬标志物LC3-II，其抗病毒活性被自噬激活剂雷帕霉素部分逆转，表明自噬抑制有助于其作用。在pedv感染的哺乳仔猪中，口服BBH （40 mg/kg/天）显著提高了仔猪的存活率（66.7%，对照组为0%），减轻了腹泻，减少了肛门粘膜的病毒脱落，降低了肠道组织（十二指肠、空肠、回肠）的病毒载量，并减轻了组织病理学损伤。总的来说，这些结果表明，BBH通过抑制病毒进入/复制和调节自噬调节来发挥有效的抗PEDV活性，支持其作为抗PEDV辅助治疗的潜力。

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引用次数: 0

Expanding the known nucleorhabdovirus world: the final chapter in a trilogy exploring the hidden diversity of plant-associated rhabdoviruses 扩展已知的核糖体病毒世界：探索植物相关横纹肌病毒隐藏多样性三部曲的最后一章。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-29 DOI: 10.1016/j.virol.2025.110699

Nicolás Bejerman , Ralf Georg Dietzgen , Humberto Debat

Nucleorhabdoviruses, classified into four distinct genera within the family Rhabdoviridae (subfamily Betarhabdovirinae), are plant-infecting viruses, characterized by nucleus-associated, enveloped, bacilliform virions. Nucleorhabdoviruses possess an unsegmented, single-stranded, negative-sense RNA genome (ca. 12–15.2 kb) that encodes six to eight proteins. Here, by exploring large publicly available metatranscriptomics datasets, we report the identification and genomic characterization of 47 novel viruses with genetic and evolutionary hallmarks of nucleorhabdoviruses. These viruses are associated with 45 distinct host plant species and were previously hidden in public database. Our findings significantly broaden the known host range of nucleorhabdoviruses, including the first nucleorhabdovirus associated with ferns and the first gammanucleorhabdoviruses linked to dicot hosts. Genetic divergence and evolutionary analyses suggest that all these novel viruses likely represent members of novel species. Phylogenetic reconstruction indicates that ten novel viruses are related to alphanucleorhabdoviruses, 25 to betanucleorhabdoviruses, eight to deltanucleorhabdoviruses and four to gammanucleorhabdoviruses. This study constitutes the final chapter in a trilogy chronicling a data mining expedition into the cryptic diversity of plant-associated rhabdoviruses, a journey that began with varicosaviruses and continued with cytorhabdoviruses. These new findings yield the most comprehensive phylogeny of nucleorhabdoviruses to date, significantly expanding our genomic understanding and illuminating the phylogenetic relationships and evolutionary dynamics of this virus group. Further, this work underscores the value of large-scale sequence data mining in advancing our understanding of the hidden world of plant rhabdoviruses.

嗜核病毒（Nucleorhabdoviruses）是一种侵染植物的病毒，其特征是与核相关的被包膜的杆菌状病毒体。核habdovirus拥有一个未分割的单链负义RNA基因组（约12-15.2 kb），可编码6 - 8种蛋白质。在这里，通过探索大量公开可用的亚转录组学数据集，我们报告了47种具有核habdov遗传和进化特征的新型病毒的鉴定和基因组特征。这些病毒与45种不同的寄主植物物种有关，以前隐藏在公共数据库中。我们的发现大大拓宽了已知的核状病毒宿主范围，包括与蕨类植物相关的第一个核状病毒和与双科宿主相关的第一个伽玛核状病毒。遗传差异和进化分析表明，所有这些新型病毒可能代表了新物种的成员。系统发育重建结果表明，10种新型病毒与α核或habdov有关，25种与β核或habdov有关，8种与δ核或habdov有关，4种与γ核或habdov有关。这项研究构成了三部曲的最后一章，记录了对植物相关横纹肌病毒隐性多样性的数据挖掘探险，这一旅程始于变异病毒，继续于细胞habdovirus。这些新发现产生了迄今为止最全面的核病毒系统发育，大大扩展了我们对基因组的理解，并阐明了该病毒群的系统发育关系和进化动力学。此外，这项工作强调了大规模序列数据挖掘在促进我们对植物横纹肌病毒隐藏世界的理解方面的价值。

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引用次数: 0

Specificity of Drosophila innubila Nudivirus infection in Drosophila cell culture 果蝇裸病毒在果蝇细胞培养中感染的特异性。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-25 DOI: 10.1016/j.virol.2025.110709

Kent M. Mulkey, Taiye S. Adewumi, Robert L. Unckless

Viral host range is an important aspect of both viral biology but also the pragmatic issue of producing viral stocks for experimentation. Host range is important both in terms of the species a virus can infect (taxonomy), and the types of host cells the virus can infect (tropism). Nudiviruses are large DNA viruses that infect several arthropods and are generally poorly studied. Several nudiviruses infect Drosophila species including the Drosophila innubila Nudivirus (DiNV). We aimed to identify cell lines that support the replication of DiNV both for the sake of understanding host range, and to develop a system for large scale production of the virus for infection. We utilized cell lines from the focal host, D. innubila, as well as available cell lines from D. virilis and D. melanogaster and inoculated with both a wild-collected pool of “naïve” virus and a D. innubila cell culture-adapted isolate. We found that virus from wild caught flies infected cells from the 3 cell lines and replicated its genome, but the passage 1 fluids from these infections were unable to reinfect upon introduction to new cells. In contrast, a cell culture-adapted strain of DiNV infected the same Drosophila cells (though relatively poorly in D. melanogaster cells) and produced infectious progeny that infected new cells. Thus, our cell culture-adapted virus developed the ability to infect broadly and produce infectious virions.

病毒宿主范围是病毒生物学的一个重要方面，也是生产用于实验的病毒库存的实际问题。宿主范围在病毒可感染的物种（分类学）和病毒可感染的宿主细胞类型（趋向性）两方面都很重要。裸病毒是一种大型的DNA病毒，可以感染几种节肢动物，对它的研究通常很少。几种裸病毒感染果蝇物种，包括果蝇nunubila裸病毒（DiNV）。我们的目标是鉴定支持DiNV复制的细胞系，以便了解宿主范围，并开发用于大规模生产感染病毒的系统。我们利用了来自焦点寄主D. innubila的细胞系，以及来自D. virilis和D. melanogaster的细胞系，并接种了野生收集的“naïve”病毒池和D. innubila细胞培养适应的分离物。我们发现来自野生捕获的苍蝇的病毒感染了3种细胞系的细胞并复制了其基因组，但这些感染的传代1流体在引入新细胞时不能再感染。相比之下，细胞培养适应的DiNV菌株感染了相同的果蝇细胞（尽管在黑腹果蝇细胞中相对较差），并产生了感染新细胞的传染性后代。因此，我们的细胞培养适应病毒发展了广泛感染和产生感染性病毒粒子的能力。

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引用次数: 0

Sequencing of the cDNA variable region of a lumpy skin disease virus ORF123 monoclonal antibody and its cross-neutralizing activity analyses against Capripoxvirus members 块状皮肤病病毒ORF123单克隆抗体cDNA可变区测序及其对卡波病毒成员的交叉中和活性分析

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-24 DOI: 10.1016/j.virol.2025.110700

Fangping Wang , Shasha Wang , Lina Tong , Huibao Wang , Haotai Chen , Xiangwei Wang , Xiangping Yin , Yuefeng Sun , Xiaolong Gao , Shanhui Ren

Monoclonal antibody sequencing is crucial for enhancing our understanding of the relationship between antibody neutralization and broad-spectrum binding. Our previous study systematically described a B-cell monoclonal antibody (mAb) derived from lumpy skin disease virus (LSDV) ORF123, which exhibits cross-reactivity with goatpoxvirus (GTPV) and sheeppoxvirus (SPPV). Here, the LSDV ORF123 mAb was sequenced for the first time using the HybSeq HT™ next-generation sequencing technique. Sequencing analyses showed that the variable heavy (V_H) and light (V_L) chains of LSDV ORF123 mAb had significant sequence similarity. Subsequently, the top 1 V_H and V_L of this LSDV ORF123 mAb were synthesized and expressed. Western blotting analyses further identified that the complementary determinant regions (CDR)1, CDR2, and CDR3 are indispensable for antigen-antibody recognition. Crucially, the co-expression of V_H + V_L of this LSDV ORF123 mAb retained its cross-neutralizing activity against LSDV, GTPV, and SPPV. Variable region sequencing of monoclonal antibodies provides a reference for improving the specificity and affinity of potential therapeutic antibody design. The cross-binding and neutralizing activity of the co-expression of V_H + V_L of the LSDV ORF123 mAb strengthens our understanding of the antigenic similarity among different members of the genus Capripoxviruses.

单克隆抗体测序对于增强我们对抗体中和和广谱结合之间关系的理解至关重要。我们之前的研究系统地描述了从疙瘩皮肤病病毒（LSDV） ORF123中提取的b细胞单克隆抗体（mAb），该抗体与山羊痘病毒（GTPV）和绵羊痘病毒（SPPV）表现出交叉反应性。本研究首次使用HybSeq HT™新一代测序技术对LSDV ORF123单抗进行测序。测序分析显示，LSDV ORF123单抗的可变重链（VH）和可变轻链（VL）具有显著的序列相似性。随后，合成并表达该LSDV ORF123单抗的前1位VH和VL。Western blotting分析进一步发现，互补决定区（CDR）1、CDR2和CDR3在抗原抗体识别中是不可或缺的。至关重要的是，该LSDV ORF123单抗的VH + VL共表达保留了其对LSDV、GTPV和SPPV的交叉中和活性。单克隆抗体的可变区测序为提高潜在治疗性抗体设计的特异性和亲和力提供了参考。LSDV ORF123单抗的VH + VL共表达的交叉结合和中和活性加强了我们对capripoxvirus属不同成员之间抗原相似性的认识。

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引用次数: 0

Corrigendum to“Neutralizing monoclonal antibodies against dengue virus: a scoping review of preclinical and clinical development” [Virology 612 (2025) 110677] “中和登革热病毒单克隆抗体：临床前和临床开发的范围审查”[病毒学612(2025)110677]的勘误表。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-22 DOI: 10.1016/j.virol.2025.110696

Irene Terzi , Dimitrios Dimitriadis , Melina Ntoga , Vasilios Petrakis , Ioulia Dragoumani , Filothei Markatou , Petros Rafailidis

引用次数: 0

A chiropteran factor of innate immunity can be recruited for rapid rescue and amplification of recombinant modified vaccinia Ankara (MVA) 重组修饰安卡拉牛苗（MVA）的快速抢救和扩增可获得一种先天性免疫翼类因子

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-19 DOI: 10.1016/j.virol.2025.110698

Aurora Elhazaz Fernandez , Sirine Abidi , Karoline Mähl, Dominique Scheffler , Volker Sandig, Ingo Jordan

Poxviruses are investigated as vectors for prophylactic and therapeutic vaccination. Transgenes are usually inserted via homologous recombination, and the desired vectors are isolated through sequential steps of plaque purification or limiting dilution. However, obtaining recombinants from the vast background of parental viruses is challenging for the ambitious timelines in personalised medicine and pandemic preparedness. We have developed a selection system based on tetherin from bats that eliminates parental viruses within the first passage after recombination. Tetherins are late-acting defensive factors that crosslink budding viruses to cellular membranes. Poxvirus morphogenesis is completed in the cytoplasm and only a subset of mature infectious particles acquires additional membranes from the trans-Golgi network. The outer of these membranes fuses with the plasma membrane for egress and is left behind. Tetherins have therefore not been suspected to interfere with release of poxviruses. We now describe that tetherin from the common vampire bat is as effective as tetherin from the Egyptian fruit bat and compare our results to conventional marker restoration based on a transiently deleted E3L gene. We demonstrate highly efficient simultaneous insertion of two transgenes into different positions of the genome of modified vaccinia Ankara (MVA) using the two tetherins. We furthermore show that recombination, rescue and amplification of vectors in suspension cultures in chemically-defined media is possible, a potential benefit for GMP campaigns. Observations with an EGFP fusion protein that is localised in the perinuclear space suggest that the bat tetherins may be active as a crosslinker during early steps of morphogenesis.

研究痘病毒作为预防和治疗性疫苗接种的载体。转基因通常通过同源重组插入，所需载体通过空斑纯化或限制稀释的连续步骤分离。然而，从庞大的亲本病毒背景中获得重组体，对于个性化医疗和大流行防范的雄心勃勃的时间表来说是一项挑战。我们已经开发了一种基于蝙蝠tetherin的选择系统，可以在重组后的第一次传代内消除亲本病毒。连接蛋白是将出芽病毒交联到细胞膜上的后效防御因子。痘病毒的形态发生在细胞质中完成，只有一小部分成熟的感染性颗粒从反式高尔基网络中获得额外的膜。这些膜的外层与质膜融合以出口并留下。因此，没有人怀疑tetherin会干扰痘病毒的释放。我们现在描述了来自普通吸血蝙蝠的tetherin与来自埃及果蝠的tetherin一样有效，并将我们的结果与基于瞬时删除的E3L基因的传统标记恢复进行了比较。我们证明了使用这两种系链蛋白将两个转基因高效地同时插入到修饰安卡拉牛苗（MVA）基因组的不同位置。我们进一步表明，在化学定义的培养基中，悬浮培养中的载体重组、拯救和扩增是可能的，这对GMP运动有潜在的好处。对位于核周空间的EGFP融合蛋白的观察表明，蝙蝠系链蛋白可能在形态发生的早期阶段作为交联剂活跃。

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引用次数: 0

The molecular mechanism and therapeutic potential of autophagy subtypes in PRRS virus infection PRRS病毒感染中自噬亚型的分子机制及治疗潜力

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-18 DOI: 10.1016/j.virol.2025.110697

Guang-Wei Zhao , Xin-Feng Li , Guo-Jia Yao , Qiu-Liang Xu

Porcine reproductive and respiratory syndrome (PRRS) virus is an RNA virus that causes reproductive failure in pregnant sows and respiratory diseases in pigs of all ages. Autophagy, a lysosome-dependent degradation process, consists of macroautophagy, microautophagy, and chaperone-mediated autophagy, which collectively maintain cellular homeostasis and influence disease progression. Although autophagy has the potential to eliminate viruses, certain viruses can manipulate this process to enhance their own replication. This review discusses the dual role of autophagy in PRRS virus infection, elucidates mechanisms underlying PRRS pathogenesis, and proposes autophagy-targeted strategies for the development of anti-PRRS virus interventions.

猪繁殖与呼吸综合征（PRRS）病毒是一种RNA病毒，可导致怀孕母猪繁殖失败和所有年龄猪的呼吸系统疾病。自噬是一种依赖溶酶体的降解过程，包括巨噬、微自噬和伴侣介导的自噬，它们共同维持细胞稳态并影响疾病进展。虽然自噬具有消灭病毒的潜力，但某些病毒可以操纵这一过程来增强自身的复制。本文综述了自噬在PRRS病毒感染中的双重作用，阐明了PRRS发病机制，并提出了以自噬为靶点的抗PRRS病毒干预策略。

引用次数: 0