Pub Date : 2025-02-01DOI: 10.1016/j.virol.2025.110392
Pei-Yu Han , Zhen-Ni Yin , Jing-Jing Yang , Meng-Si Zhang , Ying Huang , Jian Zhang , Quan-You Lu
Mulberry crinkle leaf virus (MCLV) is a representative species of the genus Mulcrilevirus in the family Geminiviridae. Here, we identified an additional V6 ORF which embedded within the V4 ORF in the MCLV virion-sense strand. The expression of V6 was confirmed by analyzing the promoter activity of V6 ORF upstream sequences and quantifying the viral DNA accumulation in V6-mutant MCLV-infected tomato plants. Infectious clones of V6-mutant MCLV vII (pCA-1.1MCLVmv6) and V4 and V6 double-mutant MCLV vII (pCA-1.1MCLVmV4mV6) were constructed, and experiments transfecting protoplasts isolated from Nicotiana benthamiana and infecting tomato using the constructs were performed. The results indicate that V6 plays a critical role in the multiplication of MCLV in protoplasts, although it has minimal, if any, impact on the systemic infection of MCLV in plants. Additionally, V4 and V6 positively regulate the MCLV DNA replication in synergistic form.
桑树皱叶病毒（Mulberry crcrickleaf virus， MCLV）是双病毒科mulcrcrilvirus属的代表种。在这里，我们发现了一个额外的V6 ORF，它嵌入在MCLV病毒粒子传感链的V4 ORF中。通过分析V6 ORF上游序列的启动子活性和对V6突变体mclv感染番茄植株的病毒DNA积累量进行定量分析，证实了V6的表达。构建了V6突变体MCLV vII （pCA-1.1MCLVmv6）和V4、V6双突变体MCLV vII （pCA-1.1MCLVmV4mV6）的侵染克隆，并利用构建体转染本烟原生质体侵染番茄进行了实验。结果表明，V6在原生质体中MCLV的增殖中起着关键作用，尽管它对MCLV在植物体内的全身感染的影响很小。此外，V4和V6以协同形式正向调节MCLV DNA复制。
{"title":"V6 encoded by mulberry crinkle leaf virus is important for viral DNA replication","authors":"Pei-Yu Han , Zhen-Ni Yin , Jing-Jing Yang , Meng-Si Zhang , Ying Huang , Jian Zhang , Quan-You Lu","doi":"10.1016/j.virol.2025.110392","DOIUrl":"10.1016/j.virol.2025.110392","url":null,"abstract":"<div><div>Mulberry crinkle leaf virus (MCLV) is a representative species of the genus <em>Mulcrilevirus</em> in the family <em>Geminiviridae</em>. Here, we identified an additional V6 ORF which embedded within the V4 ORF in the MCLV virion-sense strand. The expression of V6 was confirmed by analyzing the promoter activity of V6 ORF upstream sequences and quantifying the viral DNA accumulation in V6-mutant MCLV-infected tomato plants. Infectious clones of V6-mutant MCLV vII (pCA-1.1MCLV<sup>mv6</sup>) and V4 and V6 double-mutant MCLV vII (pCA-1.1MCLV<sup>mV4mV6</sup>) were constructed, and experiments transfecting protoplasts isolated from <em>Nicotiana benthamiana</em> and infecting tomato using the constructs were performed. The results indicate that V6 plays a critical role in the multiplication of MCLV in protoplasts, although it has minimal, if any, impact on the systemic infection of MCLV in plants. Additionally, V4 and V6 positively regulate the MCLV DNA replication in synergistic form.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110392"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142967259","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Porcine reproductive and respiratory syndrome virus (PRRSV) is a viral infectious disease that can cause infection in pigs of different ages. The condition known as porcine reproductive and respiratory syndrome poses a serious risk to the world's pig business and results in significant financial losses. Fuzhengjiedu San (FZJDS) is a traditional Chinese medicine compound, the main components include:Radix Isatidis, Radix Astragali and Herba Epimedii. It has been widely used in clinical and experimental studies, showing a wide range of biological activity. However, it is not clear whether FZJDS has anti-PRRSV activity. We observed that FZJDS had significant antiviral activity in Marc-145 cells. And FZJDS could inhibit viral infection in the stages of viral internalization and replication. Furthermore, FZJDS can inhibit PRRSV replication by inhibiting the p53 signaling pathway to affect autophagy, and FZJDS can also inhibit PRRSV replication by inhibiting the PI3K/Akt pathway.We showed in this work that FZJDS inhibits PRRSV replication in vitro and offers a novel therapeutic approach for PRRSV infection.
{"title":"Antiviral mechanism of Fuzhengjiedu San against porcine reproductive and respiratory syndrome virus","authors":"Xin-Qi Song, Xin-Yi Zhao, Wen-Shuang Chen, Li Yang, Dong-Yu Liu, Ya-Ping Chen","doi":"10.1016/j.virol.2024.110382","DOIUrl":"10.1016/j.virol.2024.110382","url":null,"abstract":"<div><div>Porcine reproductive and respiratory syndrome virus (PRRSV) is a viral infectious disease that can cause infection in pigs of different ages. The condition known as porcine reproductive and respiratory syndrome poses a serious risk to the world's pig business and results in significant financial losses. Fuzhengjiedu San (FZJDS) is a traditional Chinese medicine compound, the main components include:<em>Radix Isatidis</em>, <em>Radix Astragali</em> and <em>Herba Epimedii</em>. It has been widely used in clinical and experimental studies, showing a wide range of biological activity. However, it is not clear whether FZJDS has anti-PRRSV activity. We observed that FZJDS had significant antiviral activity in Marc-145 cells. And FZJDS could inhibit viral infection in the stages of viral internalization and replication. Furthermore, FZJDS can inhibit PRRSV replication by inhibiting the p53 signaling pathway to affect autophagy, and FZJDS can also inhibit PRRSV replication by inhibiting the PI3K/Akt pathway.We showed in this work that FZJDS inhibits PRRSV replication <em>in vitro</em> and offers a novel therapeutic approach for PRRSV infection.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110382"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142974009","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.virol.2024.110340
Marija Rozman , Laura Prtorić , Ante Šokota , Kristian Bodulić , Goran Tešović , Snjezana Zidovec-Lepej
The molecular diversity of Epstein-Barr virus (EBV) is defined by mutations in specific EBV genes and has been insufficiently studied in infectious mononucleosis (IM). The aim of this study was to determine all variations of the EBV latency genes EBNA-1, EBNA-2 and LMP-1 in pediatric patients with EBV-associated IM in Croatia, including previously defined SNPs and indels as well as previously undocumented polymorphisms. The vast majority of EBV isolates (71/72) were determined as EBV type 1 while EBNA-1 genes were classified exclusively as previously defined EBNA-1 prototypes, with 22/72 sequences categorized as P-Ala and 50/72 sequences as P-Thr. The most common LMP-1 variants included wild type (B95-8, 20/72), China1 (19/72) and recombinants (10/72). This study also described a previously undocumented polymorphism in the Arg594Lys substitution that is present in all EBNA-1 sequences examined. In addition, we found a Leu212 insertion in the EBNA-2 sequences of 50/72 isolates compared to the wild type. These polymorphisms were described for the first time in this geographic region and were not mentioned in previous studies on EBV diversity in IM. We also concluded mutual variant association between the variants using a chi-square test, in which the LMP-1 North Carolina variant was significantly more likely to appear with the EBNA-1 P-Ala prototype, while the B95-8 LMP-1 variant was significantly more likely to appear with the EBNA-1 P-Thr prototype (p < 0.05). Furthermore, leucine addition in EBNA-2 sequences is more likely to appear with LMP-1 wild type and EBNA-1 P-Thr prototype while EBV type 1 identical to the reference sequence is more likely to appear with North Carolina LMP-1 variant and EBNA-1 P-Ala prototype (p < 0.05).
eb病毒（EBV）的分子多样性是由特定EBV基因的突变决定的，在感染性单核细胞增多症（IM）中尚未得到充分的研究。本研究的目的是确定克罗地亚EBV相关IM儿科患者中EBV潜伏期基因EBNA-1、EBNA-2和LMP-1的所有变异，包括先前定义的snp和indel以及先前未记录的多态性。绝大多数EBV分离株（71/72）被确定为EBV 1型，而EBNA-1基因被完全归类为先前定义的EBNA-1原型，其中22/72序列被归类为P-Ala， 50/72序列被归类为P-Thr。最常见的LMP-1变异包括野生型（b95 - 8,20 /72）、China1（19/72）和重组型（10/72）。该研究还描述了先前未记载的在所有EBNA-1序列中存在的Arg594Lys替换多态性。此外，与野生型相比，我们在50/72分离株的EBNA-2序列中发现了一个Leu212插入。这些多态性首次在该地理区域被描述，在之前的研究中未被提及。我们还使用卡方检验得出了变体之间的相互变异关联，其中LMP-1北卡罗莱纳变体明显更有可能与EBNA-1 p - ala原型一起出现，而B95-8 LMP-1变体明显更有可能与EBNA-1 p - thr原型一起出现(p
{"title":"Previously unreported Arg594Lys in EBNA-1 and Leu212 in EBNA-2 among patients with EBV-associated infectious mononucleosis in Croatia","authors":"Marija Rozman , Laura Prtorić , Ante Šokota , Kristian Bodulić , Goran Tešović , Snjezana Zidovec-Lepej","doi":"10.1016/j.virol.2024.110340","DOIUrl":"10.1016/j.virol.2024.110340","url":null,"abstract":"<div><div>The molecular diversity of Epstein-Barr virus (EBV) is defined by mutations in specific EBV genes and has been insufficiently studied in infectious mononucleosis (IM). The aim of this study was to determine all variations of the EBV latency genes <em>EBNA-1</em>, <em>EBNA-2</em> and <em>LMP-1</em> in pediatric patients with EBV-associated IM in Croatia, including previously defined SNPs and indels as well as previously undocumented polymorphisms. The vast majority of EBV isolates (71/72) were determined as EBV type 1 while <em>EBNA-1</em> genes were classified exclusively as previously defined <em>EBNA-1</em> prototypes, with 22/72 sequences categorized as P-Ala and 50/72 sequences as P-Thr. The most common <em>LMP-1</em> variants included wild type (B95-8, 20/72), China1 (19/72) and recombinants (10/72). This study also described a previously undocumented polymorphism in the Arg594Lys substitution that is present in all <em>EBNA-1</em> sequences examined. In addition, we found a Leu212 insertion in the <em>EBNA-2</em> sequences of 50/72 isolates compared to the wild type. These polymorphisms were described for the first time in this geographic region and were not mentioned in previous studies on EBV diversity in IM. We also concluded mutual variant association between the variants using a chi-square test, in which the <em>LMP-1</em> North Carolina variant was significantly more likely to appear with the <em>EBNA-1</em> P-Ala prototype, while the B95-8 <em>LMP-1</em> variant was significantly more likely to appear with the <em>EBNA-1</em> P-Thr prototype (p < 0.05). Furthermore, leucine addition in <em>EBNA-2</em> sequences is more likely to appear with <em>LMP-1</em> wild type and <em>EBNA-1</em> P-Thr prototype while EBV type 1 identical to the reference sequence is more likely to appear with North Carolina <em>LMP-1</em> variant and <em>EBNA-1</em> P-Ala prototype (p < 0.05).</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110340"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142796751","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Begomovirus/betasatellite disease complex significantly threatens global crop production. Identifying potential plant antiviral genes is crucial for disease control. Nicotiana benthamiana is susceptible to viruses and contains 266 ethylene response transcription factors (ERFs). This study identified 29 NbERFs that were differentially upregulated in tobacco curly shoot virus and its associated betasatellite (TbCSV/TbCSB) infection, with ERF5 being the most common. Nine NbERF5s cluster phylogenetically and Niben101Scf00163g22002 (NbERF5) responds significantly to exogenous ACC treatment. Further analysis confirms the nuclear localization and transcriptional activation activity of NbERF5. Protein interaction assays demonstrate that NbERF5 has no self-interaction and does not interact with the βC1 protein of TbCSB. Silencing NbERF5 enhances TbCSV/TbCSB infection, and overexpression of NbERF5 inhibits TbCSV/TbCSB infection. Importantly, NbERF5 positively regulates the expression of the pathogenesis-related (PR) genes, NbPR1a and NbNPR1. Our findings suggest that NbERF5 enhances TbCSV/TbCSB resistance by activating the PR genes, indicating that NbERF5 is a potential antiviral gene.
{"title":"Ethylene response transcription factor 5 (ERF5) enhances defense against tobacco curly shoot virus and associated betasatellite (TbCSV/TbCSB) in Nicotiana benthamiana","authors":"Meisheng Zhao , Liping Zhang , Hussein Ghanem , Gentu Wu , Mingjun Li , Ling Qing","doi":"10.1016/j.virol.2024.110309","DOIUrl":"10.1016/j.virol.2024.110309","url":null,"abstract":"<div><div>Begomovirus/betasatellite disease complex significantly threatens global crop production. Identifying potential plant antiviral genes is crucial for disease control. <em>Nicotiana benthamiana</em> is susceptible to viruses and contains 266 ethylene response transcription factors (ERFs). This study identified 29 <em>NbERF</em>s that were differentially upregulated in tobacco curly shoot virus and its associated betasatellite (TbCSV/TbCSB) infection, with ERF5 being the most common. Nine NbERF5s cluster phylogenetically and <em>Niben101Scf00163g22002</em> (<em>NbERF5</em>) responds significantly to exogenous ACC treatment. Further analysis confirms the nuclear localization and transcriptional activation activity of NbERF5. Protein interaction assays demonstrate that NbERF5 has no self-interaction and does not interact with the βC1 protein of TbCSB. Silencing <em>NbERF5</em> enhances TbCSV/TbCSB infection, and overexpression of <em>NbERF5</em> inhibits TbCSV/TbCSB infection. Importantly, <em>NbERF5</em> positively regulates the expression of the pathogenesis-related (<em>PR</em>) genes, <em>NbPR1a</em> and <em>NbNPR1</em>. Our findings suggest that <em>NbERF5</em> enhances TbCSV/TbCSB resistance by activating the <em>PR</em> genes, indicating that <em>NbERF5</em> is a potential antiviral gene.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110309"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142824999","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.virol.2024.110380
Berenice Calderón-Pérez , Leandro Alberto Núñez-Muñoz , Lady Laura Trejo-Ayala , Víctor Hugo Rosales-García , Benjamín Emmanuel Chávez-Álvarez , Brenda Yazmín Vargas-Hernández , José Abrahán Ramírez-Pool , Roberto Ruiz-Medrano , Beatriz Xoconostle-Cázares
COVID-19 infections continue due to accessibility barriers to vaccines and the emergence of SARS-CoV-2 variants. An effective, safe, accessible, and broad-spectrum vaccine is still needed to control the disease. We developed a multivalent protein subunit vaccine comprising antigens designed from a non-N-glycosylated region of the receptor-binding domain of the spike protein of SARS-CoV-2. We combined a previously developed antigen based on the Wuhan original viral strain, and a site-mutated antigen based on several variants including Alpha, Beta, Gamma, Eta, Iota, Theta, Zeta, Mu and Omicron. The recombinant antigens were expressed in a prokaryotic system and the immunogenicity of the multivalent vaccine was tested in a mouse model. The evaluation of the subunit vaccine candidate, incorporating different variant-based multivalent recombinant antigens from non-glycosylated regions of the RBD, demonstrated a favorable safety profile, significant immunogenicity, and potent neutralizing activity, collectively supporting its potential efficacy and safety for further development.
由于疫苗可及性障碍和SARS-CoV-2变体的出现，COVID-19感染仍在继续。仍然需要一种有效、安全、可获得和广谱的疫苗来控制这种疾病。我们开发了一种多价蛋白亚单位疫苗，包括从SARS-CoV-2刺突蛋白受体结合域的非n-糖基化区域设计的抗原。我们结合了先前基于武汉原始病毒株开发的抗原，以及基于几种变体（包括Alpha， Beta, Gamma, Eta, Iota, Theta, Zeta， Mu和Omicron）的位点突变抗原。重组抗原在原核系统中表达，并在小鼠模型中测试了多价疫苗的免疫原性。对亚单位候选疫苗的评估，包括来自RBD非糖基化区域的不同基于变体的多价重组抗原，显示出良好的安全性，显著的免疫原性和有效的中和活性，共同支持其进一步开发的潜在有效性和安全性。
{"title":"Immunogenicity of a multivalent protein subunit vaccine based on non-glycosylated RBD antigens of SARS-cov-2 and its variants","authors":"Berenice Calderón-Pérez , Leandro Alberto Núñez-Muñoz , Lady Laura Trejo-Ayala , Víctor Hugo Rosales-García , Benjamín Emmanuel Chávez-Álvarez , Brenda Yazmín Vargas-Hernández , José Abrahán Ramírez-Pool , Roberto Ruiz-Medrano , Beatriz Xoconostle-Cázares","doi":"10.1016/j.virol.2024.110380","DOIUrl":"10.1016/j.virol.2024.110380","url":null,"abstract":"<div><div>COVID-19 infections continue due to accessibility barriers to vaccines and the emergence of SARS-CoV-2 variants. An effective, safe, accessible, and broad-spectrum vaccine is still needed to control the disease. We developed a multivalent protein subunit vaccine comprising antigens designed from a non-N-glycosylated region of the receptor-binding domain of the spike protein of SARS-CoV-2. We combined a previously developed antigen based on the Wuhan original viral strain, and a site-mutated antigen based on several variants including Alpha, Beta, Gamma, Eta, Iota, Theta, Zeta, Mu and Omicron. The recombinant antigens were expressed in a prokaryotic system and the immunogenicity of the multivalent vaccine was tested in a mouse model. The evaluation of the subunit vaccine candidate, incorporating different variant-based multivalent recombinant antigens from non-glycosylated regions of the RBD, demonstrated a favorable safety profile, significant immunogenicity, and potent neutralizing activity, collectively supporting its potential efficacy and safety for further development.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110380"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142901393","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.virol.2025.110405
Mohamed S.H. Hassan , Shayan Sharif
Chickens are a key species in both the manifestation of avian influenza and the potential for zoonotic transmission. Avian influenza virus (AIV) infection in chickens can range from asymptomatic or mild disease with low pathogenic AIVs (LPAIVs) to systemic fatal disease with high pathogenic AIVs (HPAIVs). During AIV infection in chickens, Toll-like receptor 7 and melanoma differentiation-associated gene 5 are upregulated to detect the single-stranded ribonucleic acid genomes of AIV, triggering a signaling cascade that produces interferons (IFNs) and pro-inflammatory cytokines. These inflammatory mediators induce the expression of antiviral proteins and recruit immune system cells, such as macrophages and dendritic cells, to the infection site. AIV evades these antiviral responses primarily through its non-structural protein 1, which suppresses type I IFNs, influencing viral pathogenicity. The uncontrolled release of pro-inflammatory cytokines may contribute to the pathogenicity and high mortality associated with HPAIV infections. AIV modulates apoptosis in chicken cells to enhance its replication, with variations in apoptosis pathways influenced by viral strain and host cell type. The presentation of AIV antigens to T and B cells leads to the production of neutralizing antibodies and the targeted destruction of infected cells by CD8+ T cells, respectively, which enhances protection and establishes immunological memory. This review explores the diverse innate and adaptive immune responses in chickens to different AIVs, focusing on the dynamics of these responses relative to protection, susceptibility, and potential immunopathology. By understanding these immune mechanisms, informed strategies for controlling AIV infection and improving chicken health can be developed.
{"title":"Immune responses to avian influenza viruses in chickens","authors":"Mohamed S.H. Hassan , Shayan Sharif","doi":"10.1016/j.virol.2025.110405","DOIUrl":"10.1016/j.virol.2025.110405","url":null,"abstract":"<div><div>Chickens are a key species in both the manifestation of avian influenza and the potential for zoonotic transmission. Avian influenza virus (AIV) infection in chickens can range from asymptomatic or mild disease with low pathogenic AIVs (LPAIVs) to systemic fatal disease with high pathogenic AIVs (HPAIVs). During AIV infection in chickens, Toll-like receptor 7 and melanoma differentiation-associated gene 5 are upregulated to detect the single-stranded ribonucleic acid genomes of AIV, triggering a signaling cascade that produces interferons (IFNs) and pro-inflammatory cytokines. These inflammatory mediators induce the expression of antiviral proteins and recruit immune system cells, such as macrophages and dendritic cells, to the infection site. AIV evades these antiviral responses primarily through its non-structural protein 1, which suppresses type I IFNs, influencing viral pathogenicity. The uncontrolled release of pro-inflammatory cytokines may contribute to the pathogenicity and high mortality associated with HPAIV infections. AIV modulates apoptosis in chicken cells to enhance its replication, with variations in apoptosis pathways influenced by viral strain and host cell type. The presentation of AIV antigens to T and B cells leads to the production of neutralizing antibodies and the targeted destruction of infected cells by CD8<sup>+</sup> T cells, respectively, which enhances protection and establishes immunological memory. This review explores the diverse innate and adaptive immune responses in chickens to different AIVs, focusing on the dynamics of these responses relative to protection, susceptibility, and potential immunopathology. By understanding these immune mechanisms, informed strategies for controlling AIV infection and improving chicken health can be developed.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110405"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143019336","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The replication and mortality caused by the viral hemorrhagic septicemia virus (VHSV) in fish vary depending on temperature. VHSV causes mortality at the temperatures below 15 °C, while infection is not established in olive flounder at temperatures above 25 °C. However, how VHSV infection manifests at the cellular level under different temperature conditions is not understood. In this study, we aimed to elucidate the mechanism by which VHSV infection is controlled by comparing viral replication and immune responses in vitro and in vivo. In the in vitro experiment, viral mRNA levels on day 5 post-challenge differed more than 10-fold between temperatures, highest at 15 °C, followed by 20 °C, and lowest at 25 °C, with replication gradually increasing over 5 days. In vivo, replication peaked by day 2 and then declined at all temperatures. VHSV infection showed a controlled tendency in the fish, whereas the virus exhibited a continuously increasing infection pattern for up to 5 days in olive flounder spleen cell cultures at a temperature of 25 °C. The level of viral infection within the cells (25 °C) was lower compared to conditions below 20 °C; however, the expression levels of interferon-related genes and pro-inflammatory cytokine genes were relatively low. The expression of ISG15 and Mx genes in spleen (25 °C) was significantly high at 24 h post challenge in the in vivo experiment.
{"title":"Temperature-dependent viral hemorrhagic septicemia virus (VHSV) infection kinetics and immune response in primary olive flounder spleen cell culture and the host","authors":"Yo-Seb Jang , Su-Young Yoon , Rahul Krishnan , Myung-Joo Oh","doi":"10.1016/j.virol.2025.110390","DOIUrl":"10.1016/j.virol.2025.110390","url":null,"abstract":"<div><div>The replication and mortality caused by the viral hemorrhagic septicemia virus (VHSV) in fish vary depending on temperature. VHSV causes mortality at the temperatures below 15 °C, while infection is not established in olive flounder at temperatures above 25 °C. However, how VHSV infection manifests at the cellular level under different temperature conditions is not understood. In this study, we aimed to elucidate the mechanism by which VHSV infection is controlled by comparing viral replication and immune responses <em>in vitro</em> and <em>in vivo</em>. In the <em>in vitro</em> experiment, viral mRNA levels on day 5 post-challenge differed more than 10-fold between temperatures, highest at 15 °C, followed by 20 °C, and lowest at 25 °C, with replication gradually increasing over 5 days. <em>In vivo</em>, replication peaked by day 2 and then declined at all temperatures. VHSV infection showed a controlled tendency in the fish, whereas the virus exhibited a continuously increasing infection pattern for up to 5 days in olive flounder spleen cell cultures at a temperature of 25 °C. The level of viral infection within the cells (25 °C) was lower compared to conditions below 20 °C; however, the expression levels of interferon-related genes and pro-inflammatory cytokine genes were relatively low. The expression of ISG15 and Mx genes in spleen (25 °C) was significantly high at 24 h post challenge in the <em>in vivo</em> experiment.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110390"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143082894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.virol.2024.110358
Da-Hye Lee , Hyo Jung Ju , Yoojin Lee , Young-Kyung Bae
Norovirus is a highly virulent pathogen that causes enteritis in all age groups worldwide. Owing to the diversity of noroviruses, the development of vaccines and treatments is challenging, and an early and accurate diagnosis is crucial. Reference materials (RMs) developed previously for norovirus genotypes I (GI) and II (GII) were quantified using reverse transcription quantitative PCR. In this study, we developed norovirus GI and GII RMs as in vitro transcribed RNA forms. These RMs were then assigned reference values for the RNA copy number concentration. The concentrations of GI and GII RMs determined using in-house reverse transcription digital PCR assays were and copy/mL, respectively. The homogeneity and stability of the RMs were evaluated, and their compatibility with commercial diagnostic kits was validated. These RMs can be used for the development of detection assays, as calibrants for various molecular measurement techniques, and as test materials for internal and external quality assurance.
{"title":"Development of RNA reference materials for norovirus GI and GII using digital PCR","authors":"Da-Hye Lee , Hyo Jung Ju , Yoojin Lee , Young-Kyung Bae","doi":"10.1016/j.virol.2024.110358","DOIUrl":"10.1016/j.virol.2024.110358","url":null,"abstract":"<div><div>Norovirus is a highly virulent pathogen that causes enteritis in all age groups worldwide. Owing to the diversity of noroviruses, the development of vaccines and treatments is challenging, and an early and accurate diagnosis is crucial. Reference materials (RMs) developed previously for norovirus genotypes I (GI) and II (GII) were quantified using reverse transcription quantitative PCR. In this study, we developed norovirus GI and GII RMs as <em>in vitro</em> transcribed RNA forms. These RMs were then assigned reference values for the RNA copy number concentration. The concentrations of GI and GII RMs determined using in-house reverse transcription digital PCR assays were <span><math><mrow><mrow><mo>(</mo><mrow><mn>1.92</mn><mspace></mspace><mo>±</mo><mspace></mspace><mn>0.37</mn></mrow><mo>)</mo></mrow><mo>×</mo><msup><mn>10</mn><mn>7</mn></msup></mrow></math></span> and <span><math><mrow><mrow><mo>(</mo><mrow><mn>1.20</mn><mo>±</mo><mspace></mspace><mn>0.27</mn></mrow><mo>)</mo></mrow><mo>×</mo><msup><mn>10</mn><mn>7</mn></msup></mrow></math></span> copy/mL, respectively. The homogeneity and stability of the RMs were evaluated, and their compatibility with commercial diagnostic kits was validated. These RMs can be used for the development of detection assays, as calibrants for various molecular measurement techniques, and as test materials for internal and external quality assurance.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110358"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857415","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
More than 95% of adult humans worldwide are latently infected with Epstein-Barr virus (EBV). Recent studies indicated that different EBV strains colonize different regions of Asia, where nasopharyngeal carcinoma (NPC) is endemic (southern China) or non-endemic (Japan/Korea). We searched for viral single nucleotide variant markers throughout the EBV genome by comparing the coding sequences of Japanese/Korean and NPC-endemic Chinese strains. We identified BamHI D fragment leftward reading frame 1 (BDLF1), BDLF2, and BDLF3 genes as viral geographical markers for distinguishing Japanese/Korean EBV strains from NPC-endemic Chinese EBV strains. Most significantly, BDLF-based EBV genotyping indicated that NPC-non-endemic Chinese and Mongolian EBV strains belong to the same group as Japanese/Korean EBV strains. We conclude that a particular type of EBV, designated as Transeurasian EBV, is prevalent among Transeurasian language speakers (Japanese, Korean, and Mongolian populations) and NPC-non-endemic Chinese populations.
{"title":"Distinct genomic features of Transeurasian strains of Epstein-Barr virus in East Asia","authors":"Hiroshi Kitamura , Iwao Kukimoto , Misako Yajima , Kazufumi Ikuta , Kenroh Sasaki , Teru Kanda","doi":"10.1016/j.virol.2024.110359","DOIUrl":"10.1016/j.virol.2024.110359","url":null,"abstract":"<div><div>More than 95% of adult humans worldwide are latently infected with Epstein-Barr virus (EBV). Recent studies indicated that different EBV strains colonize different regions of Asia, where nasopharyngeal carcinoma (NPC) is endemic (southern China) or non-endemic (Japan/Korea). We searched for viral single nucleotide variant markers throughout the EBV genome by comparing the coding sequences of Japanese/Korean and NPC-endemic Chinese strains. We identified BamHI D fragment leftward reading frame 1 (BDLF1)<em>,</em> BDLF2, and BDLF3 genes as viral geographical markers for distinguishing Japanese/Korean EBV strains from NPC-endemic Chinese EBV strains. Most significantly, BDLF-based EBV genotyping indicated that NPC-non-endemic Chinese and Mongolian EBV strains belong to the same group as Japanese/Korean EBV strains. We conclude that a particular type of EBV, designated as Transeurasian EBV, is prevalent among Transeurasian language speakers (Japanese, Korean, and Mongolian populations) and NPC-non-endemic Chinese populations.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110359"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142901404","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.virol.2024.110374
Ping Yang , Zheng-Jian Wang , Hai-Tao Lu , Xu-Meng Feng , Jing-Long Ye , Guangchuan Wang , Cheng-Feng Qin , Qing Ye , Zhong-Yu Liu
The knowledge on the life cycle of flaviviruses is still incomplete, and no direct-acting antivirals against their infections are clinically available. Herein, by screening via a Zika virus (ZIKV) replicon assay, we found that the N-terminus of NS2A exhibited great tolerance to the insertions of different split fluorescent proteins (split-FPs). Furthermore, both ZIKV and dengue virus encoding a split-FP-tagged NS2A propagated efficiently, and the split-FP-tagged ZIKVs had good genetic stability. Robust green fluorescence was observed in the reporter cell lines infected with these viruses and the fluorescence responded to anti-flavivirus chemicals with high specificity and sensitivity. Moreover, the sites of viral RNA replication were illuminated in live cells. Interestingly, by blocking viral RNA synthesis with an NS5 inhibitor, we found a correlation between the morphological characteristics of potential replication organelles and RNA amplification, highlighting that the NS2A-tagged viruses are of great value for the in-depth understanding of flavivirus replication mechanisms.
{"title":"Imaging of viral replication in live cells by using split fluorescent protein-tagged reporter flaviviruses","authors":"Ping Yang , Zheng-Jian Wang , Hai-Tao Lu , Xu-Meng Feng , Jing-Long Ye , Guangchuan Wang , Cheng-Feng Qin , Qing Ye , Zhong-Yu Liu","doi":"10.1016/j.virol.2024.110374","DOIUrl":"10.1016/j.virol.2024.110374","url":null,"abstract":"<div><div>The knowledge on the life cycle of flaviviruses is still incomplete, and no direct-acting antivirals against their infections are clinically available. Herein, by screening via a Zika virus (ZIKV) replicon assay, we found that the N-terminus of NS2A exhibited great tolerance to the insertions of different split fluorescent proteins (split-FPs). Furthermore, both ZIKV and dengue virus encoding a split-FP-tagged NS2A propagated efficiently, and the split-FP-tagged ZIKVs had good genetic stability. Robust green fluorescence was observed in the reporter cell lines infected with these viruses and the fluorescence responded to anti-flavivirus chemicals with high specificity and sensitivity. Moreover, the sites of viral RNA replication were illuminated in live cells. Interestingly, by blocking viral RNA synthesis with an NS5 inhibitor, we found a correlation between the morphological characteristics of potential replication organelles and RNA amplification, highlighting that the NS2A-tagged viruses are of great value for the in-depth understanding of flavivirus replication mechanisms.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"603 ","pages":"Article 110374"},"PeriodicalIF":2.8,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142928836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号