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Analysis of the surface topology of respiratory syncytial virus particles that form on the surface of virus-infected cells 在病毒感染细胞表面形成的呼吸道合胞病毒颗粒的表面拓扑结构分析

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-31 DOI: 10.1016/j.virol.2025.110732

Chris E. Jeffree , Thong Beng Lu , Boon-Huan Tan , Richard J. Sugrue

The surface topology of virus filaments on respiratory syncytial virus (RSV)-infected cells was examined using field emission gun-scanning electron microscopy (FEG-SEM) and atomic force microscopy (AFM). FEG-SEM analysis of the surface of RSV-infected cells labelled with anti-F and anti-G protein antibodies revealed the presence of virus filaments and clusters of the F and G proteins distributed intermittently along their surface of the virus filaments. RSV-infected cells thinly coated with chromium were imaged using FEG-SEM and revealed a distinct structured surface topology consisting of closely packed surface domains. The G protein clusters were only associated within a subset of these domains which suggested that this structured topology was mainly derived from the host cell. Imaging of RSV-infected cells using AFM was undertaken as a different but complementary approach to the FEG-SEM analysis. Imaging using AFM revealed a similar structured surface topology on the virus filaments to that observed in the FEG-SEM analysis, indicating the consistency in the appearance of the virus surface topology using these different methods. Collectively, this study provides the first detailed imaging of the surface topology of the virus filaments as they form on RSV-infected cells. The imaging data is consistent with the envelopment of the virus filaments by cellular membrane microdomains and the concentration of the virus glycoproteins in other microdomains at specific locations on the virus filaments. Collectively, these data further highlight the complexity of the spatial organisation within the viral envelope during virus assembly.

采用场发射枪扫描电镜（FEG-SEM）和原子力显微镜（AFM）观察了呼吸道合胞病毒（RSV）感染细胞上病毒丝的表面拓扑结构。用抗F和抗G蛋白抗体标记的rsv感染细胞表面的FEG-SEM分析显示存在病毒丝和沿病毒丝表面间歇性分布的F和G蛋白簇。rsv感染的细胞被薄镀铬，使用fg - sem成像，显示出由紧密排列的表面域组成的独特结构表面拓扑结构。G蛋白簇仅在这些结构域的一个子集内相关，这表明这种结构拓扑主要来自宿主细胞。使用AFM对rsv感染的细胞进行成像，作为一种不同但补充的方法，以进行FEG-SEM分析。利用AFM成像显示，病毒细丝上的结构表面拓扑结构与在fg - sem分析中观察到的结构表面拓扑结构相似，表明使用这些不同方法的病毒表面拓扑结构外观的一致性。总的来说，这项研究首次提供了病毒丝在rsv感染细胞上形成时的表面拓扑结构的详细成像。成像数据与病毒细丝被细胞膜微结构域包膜和病毒细丝上特定位置的其他微结构域的病毒糖蛋白浓度一致。总的来说，这些数据进一步强调了病毒组装过程中病毒包膜内空间组织的复杂性。

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引用次数: 0

Molecular biology of Cotton Leafroll Dwarf Virus (CLRDV) and potential application of CRISPR-Cas technology for developing virus-resistant cotton 棉花叶卷矮病毒（CLRDV）的分子生物学特性及CRISPR-Cas技术在棉花抗病育种中的潜在应用

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-30 DOI: 10.1016/j.virol.2025.110730

Sriharsha V. Lankireddy , Saikrishna Lekkala , Archana Khadgi , Venkateswara R. Sripathi , Madhusudhana R. Janga

Cotton leafroll dwarf virus (CLRDV) poses an increasing threat to global cotton production. Transmitted by the cotton aphid (Aphis gossypii) in a persistent, circulative manner, CLRDV exhibits a wide geographical distribution, with documented presence in South America, Africa, Asia, and the USA. Infection can result in either cotton blue disease (CBD) in South America or cotton leafroll dwarf disease (CLRDD) in the USA, both of which are associated with CLRDV. The considerable genetic diversity and frequent recombination events within CLRDV populations contribute to this symptom variability and complicate both diagnosis and management. While resistant cultivars have reduced disease impact in South America, these lines remain susceptible to emerging US strains, underscoring the urgent need for region-specific resistance breeding. Current molecular diagnostics rely on RT-PCR, but there is a need for rapid, field-deployable detection tools. Recent advances, such as CRISPR-Cas13a based SHERLOCK assays, offer sensitive and specific detection of CLRDV, with potential for on-site applications. Efficient screening techniques, supported by next-generation sequencing and transcriptomics, are essential for identifying novel resistance sources and elucidating virus-host interactions. CRISPR-based genome editing holds significant promise, as demonstrated in other crops. Targeted disruption of host susceptibility genes using CRISPR-Cas9, or direct degradation of viral genomes with RNA-targeting systems such as Cas12/Cas13, could offer durable, broad-spectrum resistance. By integrating molecular virology, high-throughput genomics, and precision gene editing, this review outlines a roadmap for translating these advances into sustainable, field-level solutions for CLRDV management and long-term cotton productivity.

棉花卷叶矮缩病毒（CLRDV）对全球棉花生产的威胁日益严重。CLRDV由棉蚜（棉蚜）传播，以持久、循环的方式传播，具有广泛的地理分布，在南美洲、非洲、亚洲和美国都有记录。感染可导致南美的棉花蓝病（CBD）或美国的棉花叶卷侏儒病（CLRDD），两者都与CLRDV有关。在CLRDV人群中，相当大的遗传多样性和频繁的重组事件导致了这种症状的变异性，并使诊断和管理复杂化。虽然抗性品种减少了南美的疾病影响，但这些品系仍然容易受到美国新出现的菌株的影响，这强调了迫切需要进行区域特异性抗性育种。目前的分子诊断依赖于RT-PCR，但需要快速的、可现场部署的检测工具。最近的进展，如基于CRISPR-Cas13a的SHERLOCK检测，提供了对CLRDV的敏感和特异性检测，具有现场应用的潜力。在下一代测序和转录组学的支持下，高效的筛选技术对于鉴定新的耐药性来源和阐明病毒与宿主的相互作用至关重要。基于crispr的基因组编辑具有重大的前景，正如在其他作物中所证明的那样。使用CRISPR-Cas9靶向破坏宿主易感基因，或使用rna靶向系统（如Cas12/Cas13）直接降解病毒基因组，可以提供持久的广谱抗性。通过整合分子病毒学、高通量基因组学和精确基因编辑，本文概述了将这些进展转化为CLRDV管理和棉花长期生产力可持续的田间解决方案的路线图。

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引用次数: 0

Differential induction of PD-L1 expression in cells infected with feline infectious peritonitis virus and feline enteric coronavirus 猫传染性腹膜炎病毒和猫肠道冠状病毒感染细胞PD-L1表达的差异诱导

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-30 DOI: 10.1016/j.virol.2025.110734

Alexandria Zabiegala, Yunjeong Kim, Kyeong-Ok Chang

Feline infectious peritonitis (FIP) is a fatal systemic disease of cats, which is caused by FIP virus (FIPV), a virulent biotype of feline coronavirus (FCoV). In a small number of cats infected with feline enteric coronavirus (FECV), virus mutation may emerge that enable infection of monocytes and macrophages, leading to the development of FIP. This cell tropism shift, along with impaired T cell response, plays a critical role in FIP development. Programmed cell death protein (PD-1) and its ligands (PD-L1 and PD-L2) play crucial roles in immune responses as an immune checkpoint system. The PD-1 axis downregulates T cell response and is reported to be involved in immune evading mechanisms of various viruses. In this study, we used two virus strains FIPV-1146 and FECV-1683 corresponding to FIPV and FECV biotypes, respectively, to study the role of PD-L1 expression as a potential mechanism for FIP development. While both viruses demonstrated robust viral replication at comparable levels, FIPV-1146, but not FECV-1683, significantly upregulated PD-L1 expression in CRFK and and Fcwf-4 cells. Infection of Fcwf-4 cells with FIPV-Black strain also increased PD-L1 levels. Co-culture studies using Jurkat cells and Fcwf-4 cells infected with FIPV-1146 or FECV-1683 showed that PD-L1 induction by FIPV-1146 attenuated T cell activation. The cell signaling profiling assays showed that FIPV-1146, but not FECV-1683, induced the type I IFN synthesis, which is a well-known regulator for PD-L1 expression. The elevated PD-L1 levels by FIPV may lead to dampened T cell response, allowing persistent infection in macrophages and development of the FIP clinical disease.

猫传染性腹膜炎（FIP）是由猫冠状病毒（FCoV）的一种强毒生物型FIP病毒（FIPV）引起的猫的一种致死性全身性疾病。在少数感染猫肠道冠状病毒（FECV）的猫中，可能出现病毒突变，使单核细胞和巨噬细胞感染，导致FIP的发展。这种细胞向性转变，以及受损的T细胞反应，在FIP的发展中起着关键作用。程序性细胞死亡蛋白（PD-1）及其配体（PD-L1和PD-L2）作为免疫检查点系统在免疫应答中起着至关重要的作用。PD-1轴下调T细胞应答，据报道参与多种病毒的免疫逃避机制。本研究利用FIPV和FECV生物型分别对应的两种病毒株FIPV-1146和FECV-1683，研究PD-L1表达作为FIP发生的潜在机制的作用。虽然这两种病毒在相当水平上表现出强大的病毒复制，但fipv1146，而FECV-1683，在CRFK和Fcwf-4细胞中显著上调PD-L1表达。FIPV-Black菌株感染Fcwf-4细胞后，PD-L1水平升高。用FIPV-1146或FECV-1683感染Jurkat细胞和Fcwf-4细胞共培养研究表明，FIPV-1146诱导PD-L1可减弱T细胞的活化。细胞信号分析分析显示，fiv -1146而非FECV-1683诱导I型IFN合成，而I型IFN是众所周知的PD-L1表达调节剂。FIPV引起的PD-L1水平升高可能导致T细胞反应减弱，从而导致巨噬细胞持续感染和FIP临床疾病的发展。

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引用次数: 0

Viral diversity in vertebrates from Alto Pantanal, Mato Grosso, 2019 马托格罗索州上潘塔纳尔脊椎动物的病毒多样性，2019

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-30 DOI: 10.1016/j.virol.2025.110729

Janeth Aracely Ramirez Pavon , Nilvanei Aparecido da Silva Neves , Anderson de Oliveira Martins , João Batista Pinho , Vilma Juscineide de Souza , Sandro Patroca da Silva , Ana Cecília Ribeiro Cruz , Daniele Barbosa de Almeida Medeiros , Márcio Roberto Teixeira Nunes , Renata Dezengrini Slhessarenko

The Alto Pantanal is a key yet unexplored Brazilian wetland for studying vertebrate viral profiles. This study presents viral taxonomic profiles from vertebrate sera sampled in May, June and October of 2019 in Porto São Luiz and Pirizal, Alto Pantanal. A total of 13 frogs, 19 bats, 23 caimans, 36 equids, 20 domestic hens, 140 birds and 16 humans were sampled. After nucleic acid extraction, individual samples were pooled by species and subjected to a metagenomic approach. Viral reads accounted for less than 0.2 % in each pool, except in domestic hens (35.8 %), yielding an overall viral abundance variation among pools of 48.83 %. In total, twenty-nine viral genomic sequences were retrieved from five pools. In equids, two coding-complete genomes were identified belonging to species Copiparvovirus ungulate8 and Mutorquevirus equid2. In domestic hens, four coding-complete genomes of species Alpharetrovirus avileu were detected, along with partial genomes of three gyroviroviruses of species Gyrovirus homsa1, Gyrovirus galga1 and a putative novel unclassified gyrovirus. In humans, 15 genomes of known human anelloviruses were identified, as well as partial sequences of Orthoflavivirus ilheusense and Erythroparvovirus primate species. In caimans, a partial genome belonging to genus Betadintovirus was detected. In frogs, one partial sequence of a putative novel pegivirus, and a coding-complete sequence of an unclassified retrovirus (Rhinella marina endogenous retrovirus) were found. These findings provide valuable insights into viral circulation within the diverse Pantanal biome, and support viral genomic surveillance efforts in the region.

上潘塔纳尔是研究脊椎动物病毒概况的一个尚未开发的巴西湿地。本研究展示了2019年5月、6月和10月在上潘塔纳尔港的波尔图s o Luiz和Pirizal采集的脊椎动物血清的病毒分类特征。共采集了13只青蛙、19只蝙蝠、23只凯门鳄、36只马科动物、20只家母鸡、140只鸟类和16名人类。核酸提取后，个体样本按物种汇总，并进行宏基因组分析。除家母鸡（35.8%）外，每个池的病毒读取率均低于0.2%，因此池之间的总体病毒丰度差异为48.83%。总共从五个库中检索了29个病毒基因组序列。在马科动物中，鉴定出两个编码完整的基因组，分别属于有蹄的Copiparvovirus 8和马科的mutorquvirus 2。在家鸡中检测到4个编码完整的甲型逆转录病毒（Alpharetrovirus avileu）基因组，以及3个旋转病毒（Gyrovirus homsa1、Gyrovirus galga1和一种推测为新型未分类的旋转病毒）的部分基因组。在人类中，已鉴定出15个已知的人类无绒病毒基因组，以及灵长类物种正黄病毒和红细小病毒的部分序列。在凯门鳄中，检测到一个属于betadinvirus属的部分基因组。在蛙类中，发现了一种假定的新型反转录病毒的部分序列和一种未分类的逆转录病毒（滨海鼻杆菌内源性逆转录病毒）的编码完整序列。这些发现为了解潘塔纳尔不同生物群落中的病毒循环提供了有价值的见解，并支持该地区的病毒基因组监测工作。

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引用次数: 0

The antiviral activity of interferon-stimulated genes (ISGs) in influenza A virus infection 干扰素刺激基因（ISGs）在甲型流感病毒感染中的抗病毒活性

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-28 DOI: 10.1016/j.virol.2025.110728

Aoxue Wang , Jiashu Hu , Qianqian Zhang , Yuying Zhang , Fanhua Wei

This review systematically delineates the antiviral activity of interferon-stimulated genes (ISGs) against influenza A virus (IAV) infection and their underlying mechanisms. IAV, a major respiratory pathogen, evades immune surveillance through antigenic drift and shift, leading to seasonal epidemics and pandemics. The host innate immune system recognizes IAV via pattern recognition receptors (PRRs), activating interferon (IFN) signaling pathways to induce ISG expression, thereby suppressing viral replication. ISGs target multiple stages of the IAV replication cycle through direct or indirect mechanisms. Additionally, IFN-induced non-coding RNAs reinforce antiviral defenses by modulating host genes or directly targeting viral genomes. Despite their critical roles, some viral proteins counteract ISGs, reflecting host-pathogen conflict. Further research is needed to fully elucidate ISG mechanisms, offering insights for novel antiviral strategies.

本文综述了干扰素刺激基因（ISGs）对甲型流感病毒（IAV）感染的抗病毒活性及其潜在机制。IAV是一种主要的呼吸道病原体，通过抗原漂移和转移逃避免疫监测，导致季节性流行和大流行。宿主先天免疫系统通过模式识别受体（PRRs）识别IAV，激活干扰素（IFN）信号通路诱导ISG表达，从而抑制病毒复制。isg通过直接或间接机制靶向IAV复制周期的多个阶段。此外，ifn诱导的非编码rna通过调节宿主基因或直接靶向病毒基因组来增强抗病毒防御。尽管它们具有关键作用，但一些病毒蛋白抵消isg，反映了宿主-病原体的冲突。需要进一步的研究来充分阐明ISG的机制，为新的抗病毒策略提供见解。

引用次数: 0

SARS-CoV-2 Spike displays multiple adaptive changes in addition to the furin cleavage site SARS-CoV-2 Spike除了具有furin切割位点外，还表现出多种适应性变化

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-24 DOI: 10.1016/j.virol.2025.110720

Robert F. Garry

Evolution of SARS-CoV-2 from bat sarbecoviruses involved multiple changes in Spike in addition to insertion of the furin cleavage site (FCS). Analysis of the closely related Spike of BANAL-20-52 reveals key adaptations in the SARS-CoV-2 Spike beyond the FCS that occurred prior to the spillover of SARS-CoV-2's immediate progenitor to humans. Bat sarbecoviruses have enteric tropism and spread mostly by the gastrointestinal route. Their Spike proteins predominantly assume the locked form, which is able to resist the low pH of the bat gastrointestinal tract. Initial changes during the SARS-CoV-2 evolutionary pathway included substitutions that expanded the host range of the sarbecovirus progenitor and allowed circulation in nonbat mammals. Adaptation of the SARS-CoV-2 progenitors also involved remodeling of the amino-terminal domain. Respiratory adaptation occurred during circulation in nonbat animals and resulted in greater propensity for Spike to assume open forms that are less compact and more metastable than the locked or closed forms. Substitutions at monomer interfaces in the Spike trimer facilitate the open shift. Like FCS insertion, these substitutions make Spike more susceptible to low pH degradation and could not have occurred in bats. After SARS-CoV-2 spilled over to humans Spikes of the dominant lineage acquired an aspartic acid to glycine substitution at position 614 that further decreases interaction between monomers and promotes opening of the Spike trimer. A multi-stage evolutionary trajectory is also evident during cross-species transmissions of bat sarbecoviruses to pangolins and the first known spillovers of SARS-CoV via the wildlife trade.

除了插入furin切割位点（FCS）外，蝙蝠sarbecovirus的SARS-CoV-2进化还涉及Spike的多种变化。对密切相关的BANAL-20-52刺突的分析揭示了SARS-CoV-2刺突在SARS-CoV-2的直接祖细胞向人类扩散之前发生的FCS以外的关键适应。蝙蝠sarbecovirus具有肠性，主要通过胃肠道传播。它们的刺突蛋白主要呈锁定形式，能够抵抗蝙蝠胃肠道的低pH值。在SARS-CoV-2进化途径中的初始变化包括替换，这些替换扩大了sarbecovirus祖病毒的宿主范围，并允许在非蝙蝠哺乳动物中传播。SARS-CoV-2祖细胞的适应还涉及氨基末端结构域的重塑。呼吸适应发生在非蝙蝠动物的循环过程中，导致Spike更倾向于采取开放形式，这种形式比锁定或封闭形式更不紧密，更稳定。在Spike三聚体的单体界面上的取代促进了开位移。与FCS插入一样，这些替换使Spike更容易受到低pH降解的影响，并且不可能发生在蝙蝠身上。在SARS-CoV-2扩散到人类之后，优势谱系的Spike在614位获得了天冬氨酸到甘氨酸的取代，这进一步减少了单体之间的相互作用，并促进了Spike三聚体的开放。在蝙蝠sarbecov向穿山甲的跨物种传播以及已知的sars冠状病毒通过野生动物贸易的首次溢出过程中，也明显存在多阶段的进化轨迹。

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引用次数: 0

Forsythoside A inhibits avian infectious bronchitis virus infection by binding the S1 subunit 连翘苷A通过结合S1亚基抑制禽传染性支气管炎病毒感染。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-22 DOI: 10.1016/j.virol.2025.110715

Ruiting Shen , Jinwei Guo , Xuewei Liu , Peng Yin , Xiaolin Hou

Objectives

Avian infectious bronchitis virus (IBV) causes acute, highly contagious bronchitis inflammation and severe economic losses in the poultry industry. Such infections are difficult to treat as the virus has high mutation rates. Forsythiaside A (FTA) has an apparent inhibitory effect on IBV. This article describes evidence for the activity and the exact mechanism against IBV.

Methods

We observed the inhibitory effect of FTA on IBV infection lifecycle stages using plaque counts, immunofluorescence assay, and RT‒qPCR; then synthesized FTA-OVA, and expressed the recombination protein of IBV spike protein; thirdly, constructed an enzyme-linked immunosorbent assay (ELISA) based on the interaction assay between FTA and IBV particles or IBV spike protein. The interaction between FTA and the S protein was further confirmed using an isothermal titration calorimetry (ITC) system. We predicted the bond modes between FTA and S protein by molecular Docking. Moreover, changed the potential binding amino acids. Then we compared the affinity between FTA and the S1 mutant or the S1 wild-type.

Results

FTA inhibited IBV infection, especially on the viral attachment stage at non-cytotoxic concentrations, and FTA bound to virus particles or IBV S1 protein, rather than the S2 subunit. Molecular Docking via Autodock software revealed that FTA could form hydrogen bonds with eight amino acids in the S1 subunit, and these amino acids were then mutated. The affinity between FTA and the S1 mutant was much lower than that of the S1 wild-type.

Conclusion

FTA prevents IBV infection by blocking viral absorption and contact with the S1 subunit of IBV but not with S2. We further identified the binding sites by comparing wild-type and mutant S1 affinities.

目的：禽传染性支气管炎病毒（IBV）引起急性、高度传染性支气管炎炎症，给家禽业造成严重的经济损失。这种感染很难治疗，因为病毒的突变率很高。连翘苷A （FTA）对IBV有明显的抑制作用。本文描述了该活性的证据和对抗IBV的确切机制。方法：采用菌斑计数、免疫荧光法和RT-qPCR观察FTA对IBV感染生命周期阶段的抑制作用；然后合成FTA-OVA，表达IBV刺突蛋白重组蛋白；第三，在FTA与IBV颗粒或IBV刺突蛋白相互作用的基础上，构建酶联免疫吸附试验（ELISA）。用等温滴定量热法（ITC）进一步证实了FTA与S蛋白的相互作用。通过分子对接预测了FTA与S蛋白之间的键合模式。此外，改变了潜在的结合氨基酸。然后比较了FTA与S1突变体和S1野生型的亲和性。结果：FTA对IBV感染有抑制作用，特别是在无细胞毒性浓度的病毒附着阶段，FTA与病毒颗粒或IBV S1蛋白结合，而不是与S2亚基结合。通过Autodock软件进行分子对接，发现FTA可以与S1亚基上的8个氨基酸形成氢键，这些氨基酸随后发生突变。FTA与S1突变体的亲和性远低于S1野生型。结论：FTA通过阻断病毒吸收和与IBV S1亚基的接触而不是与S2亚基的接触来阻止IBV感染。我们通过比较野生型和突变型S1的亲和力进一步确定了结合位点。

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引用次数: 0

Carica papaya genome-encoded microRNAs as post-transcriptional silencers of Papaya ringspot virus: A genome-wide mining and RT-qPCR-based functional validation 番木瓜基因组编码的microrna作为番木瓜环斑病毒转录后沉默子：全基因组挖掘和基于rt - qpcr的功能验证。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-18 DOI: 10.1016/j.virol.2025.110718

Rajkumari Jyotika , Sankarasubramanian Harish , Mannu Jayakanthan , Gandhi Karthikeyan , K.K. Kumar , Marimuthu Murugan , Santosh Ganapati Patil

MicroRNAs (miRNAs) play a crucial role in RNA interference (RNAi)-mediated antiviral defence, offering potential applications in virus detection and breeding for resistance. The present study aimed to isolate total RNA from PRSV-infected papaya plants and investigate the expression profiles of conserved miRNAs involved in papaya–PRSV interactions. Potential miRNA target sites were predicted using four complementary bioinformatics tools—miRanda, RNA22, plant small RNA (psRNA) Target, and RNAhybrid—and conserved targets were identified across PRSV isolates from Taiwan, Hawaii, and Delhi. To validate the in-silico predictions, specific stem-loop primers were designed for miRNA reverse transcription, followed by pulse RT-based quantitative real-time PCR (RT-qPCR) for precise expression profiling. Differential expression analysis revealed significant upregulation of cpa-miR159a, cpa-miR394a, cpa-miR394b, and cpa-miR319, whereas cpa-miR8140 was markedly downregulated in infected plants. These findings indicate that miRNAs may be involved in modulating host-defense responses during PRSV infection. Overall, this study provides valuable insights into the miRNA–mRNA interaction network and establishes a foundational resource that can support future research aimed at understanding the molecular mechanisms underlying PRSV–papaya interactions.

MicroRNAs （miRNAs）在RNA干扰（RNAi）介导的抗病毒防御中起着至关重要的作用，在病毒检测和抗性育种中具有潜在的应用前景。本研究旨在从prsv感染的木瓜植株中分离总RNA，并研究参与木瓜- prsv相互作用的保守mirna的表达谱。利用四种互补的生物信息学工具（miranda、RNA22、植物小RNA (psRNA) target和rnahybrid）预测潜在的miRNA靶点，并在来自台湾、夏威夷和德里的PRSV分离株中鉴定出保守的靶点。为了验证计算机预测，设计了特定的茎环引物用于miRNA反转录，然后使用基于脉冲rt的定量实时PCR （RT-qPCR）进行精确的表达谱分析。差异表达分析显示，在感染植株中，pa- mir159a、pa- mir394a、pa- mir394b和pa- mir319表达显著上调，而pa- mir8140表达显著下调。这些发现表明，在PRSV感染期间，mirna可能参与调节宿主防御反应。总的来说，这项研究为miRNA-mRNA相互作用网络提供了有价值的见解，并建立了基础资源，可以支持未来旨在了解prv -木瓜相互作用的分子机制的研究。

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引用次数: 0

Identification of immunodominant epitope on African swine fever virus K145R protein using monoclonal antibodies 非洲猪瘟病毒K145R蛋白免疫优势表位的单克隆抗体鉴定

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-17 DOI: 10.1016/j.virol.2025.110719

Weldu Tesfagaber , Lezi Hou , Wan Wang , Yuanmao Zhu , Encheng Sun , Jingli Liu , Taijie Guo , Renqiang Liu , Zhigao Bu , Fang Li , Dongming Zhao

African swine fever (ASF) is a lethal viral disease that affects domestic pigs and wild boars, resulting in significant economic consequences. The causative agent, which is a large, double-stranded DNA virus, encodes over 150 proteins, many of which have unknown functions. Limited knowledge of the antigenic properties of these proteins has hindered the development of vaccines against ASFV. Prior research has indicated that the K145R protein is immunogenic and expressed abundantly in ASFV-infected cells. In this study, we generated and characterized six monoclonal antibodies (mAbs) against the E. coli expressed K145R protein. Analysis through ELISA, Western blot, and immunofluorescent assay (IFA) demonstrated that the mAbs specifically and strongly react to the K145R protein and ASFV-infected cells. To identify the key linear B cell epitopes of the mAbs, a series of truncated recombinant K145R proteins with maltose-binding protein (MBP) tags were constructed, expressed in E. coli, and analysed. All mAbs specifically recognized a linear epitope located at the C-terminal region of K145R, spanning the sequence ¹³³TWAKIVEEG¹⁴¹. Further investigation using alanine-scanning mutagenesis revealed that tryptophan (W¹³⁴) and Alanine (A¹³⁵) are critical amino acid residues for epitope-antibody interactions. The linear epitope exhibited specific reactions with ASFV-positive serum in an indirect ELISA and was able to distinguish between ASFV positive and negative pig serum. These mAbs and their defined epitopes will provide additional information to understand the structure and immunological characteristics of K145R and lay the foundation for the development of epitope-based diagnostic tools for ASFV.

非洲猪瘟（ASF）是一种影响家猪和野猪的致命病毒性疾病，造成严重的经济后果。这种病原体是一种大型的双链DNA病毒，可以编码150多种蛋白质，其中许多蛋白质的功能未知。对这些蛋白抗原特性的有限了解阻碍了非洲猪瘟疫苗的开发。先前的研究表明，K145R蛋白具有免疫原性，并在asfv感染的细胞中大量表达。在本研究中，我们制备并鉴定了6种针对大肠杆菌表达K145R蛋白的单克隆抗体（mab）。ELISA、Western blot和免疫荧光（IFA）分析表明，单克隆抗体对K145R蛋白和asfv感染细胞具有特异性和强反应。为了鉴定单克隆抗体的关键线性B细胞表位，构建了一系列带麦芽糖结合蛋白（maltose-binding protein， MBP）标签的截断重组K145R蛋白，在大肠杆菌中表达并分析。所有单克隆抗体特异性识别位于K145R c端区域的线性表位，跨越序列133TWAKIVEEG141。进一步研究发现，色氨酸（W134）和丙氨酸（A135）是表位-抗体相互作用的关键氨基酸残基。在间接ELISA中，线性表位与ASFV阳性血清表现出特异性反应，能够区分猪ASFV阳性和阴性血清。这些单克隆抗体及其确定的表位将为了解K145R的结构和免疫学特性提供额外的信息，并为开发基于表位的ASFV诊断工具奠定基础。

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引用次数: 0

Transcriptome analysis of Crandell Rees Feline Kidney (CRFK) cells infected with Feline calicivirus strain 023 (FCV 023) 猫纹状病毒023株（FCV 023）感染CRFK细胞的转录组分析。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-10-15 DOI: 10.1016/j.virol.2025.110716

Ruiming Zhang , Hongwei Zhu , Guangrong Zhao , Xiu Xue , Xin Yu , Yang Liu , Jiayu Yu , Linlin Jiang , Jianlong Zhang , Xingxiao Zhang

Vesivirus felis (Feline calicivirus, FCV) is a widely prevalent viral pathogen in domestic cats, commonly associated with upper respiratory tract infections. However, the transcriptomic responses of host cells to FCV infection remain largely uncharacterized. In this study, an in vitro FCV infection model was established using Crandell-Rees Feline Kidney (CRFK) cells. High-throughput RNA sequencing (RNA-Seq) was performed at 4, 8, and 12 h post-infection (hpi) to investigate the dynamics of host gene expression. Differentially expressed genes (DEGs) were identified and subjected to Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein-protein interaction (PPI) network analyses. A total of 829, 5,432, and 6342 DEGs were detected at 4, 8, and 12 hpi, respectively. Key enriched pathways were associated with metabolic processes, endoplasmic reticulum stress, cytoskeletal remodeling, apoptosis, and immune responses, including the activation of Ubiquitin-mediated proteolysis, Apoptosis signaling, Adherens junctions, and the MAPK, NF-κB, and Toll-like receptor signaling pathways. This study provides a comprehensive transcriptomic landscape of FCV-infected CRFK cells and identifies key molecular mechanisms involved in virus-host interactions, offering potential targets for antiviral therapy.

猫膀胱病毒（Feline calicivirus， FCV）是一种在家猫中广泛流行的病毒性病原体，通常与上呼吸道感染有关。然而，宿主细胞对FCV感染的转录组反应在很大程度上仍未被表征。本研究采用CRFK （crandel - rees猫肾）细胞建立体外FCV感染模型。在感染后4、8和12小时进行高通量RNA测序（RNA- seq），研究宿主基因表达的动态。对差异表达基因（DEGs）进行鉴定，并进行基因本体（GO）、京都基因与基因组百科全书（KEGG）和蛋白质-蛋白质相互作用（PPI）网络分析。在4、8和12 hpi处分别检测到829、5432和6342个deg。关键富集通路与代谢过程、内质网应激、细胞骨架重塑、细胞凋亡和免疫应答相关，包括泛素介导的蛋白水解、细胞凋亡信号、粘附连接以及MAPK、NF-κB和toll样受体信号通路的激活。本研究提供了fcv感染CRFK细胞的全面转录组学景观，并确定了参与病毒-宿主相互作用的关键分子机制，为抗病毒治疗提供了潜在的靶点。

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