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Revisiting the roles of trypsin in the productive infection of porcine deltacoronavirus in porcine-derived cells

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-14 DOI: 10.1016/j.virol.2025.110453

Wenwen Xiao , Zhuang Li , Chaoqun Chen , Yuting Shi , Puxian Fang , Shaobo Xiao , Liurong Fang

Porcine deltacoronavirus (PDCoV) is an emerging enteric coronavirus with the potential for interspecies transmission. Trypsin has been shown to play a positive role in the isolation and multiplication of PDCoV in vitro, however, the functions of trypsin during PDCoV replication cycle remain controversial. In this study, we revisited the roles of trypsin for PDCoV infection by utilizing two kinds of PDCoV, PDCoV^T+ and PDCoV^T−, which were prepared in the presence or absence of trypsin, respectively. We found that PDCoV^T+ was able to continuously proliferate in the medium containing trypsin, achieving a higher titer as the infection progress in LLC-PK1 and other tested porcine-derived cells. However, its replication was only transiently improved at 12 hours post-infection, and lower viral titers were observed under trypsin-free culture conditions. Furthermore, the trypsin-mediated enhancement of viral replication could be inhibited by trypsin inhibitor SBTI, suggesting that the second-round viral reproduction of PDCoV^T⁺ might be impeded without trypsin. We further investigated the replication dynamics of PDCoV^T− in LLC-PK1 cells in the presence or absence of trypsin. The results indicated that PDCoV^T− generated lower viral titers under trypsin-free culture conditions, while the addition of trypsin reverted the infectivity of PDCoV^T−. Additionally, we demonstrated that trypsin cleaved the PDCoV spike protein, activating viral attachment and internalization. Moreover, trypsin promoted viral replication and release, accelerating PDCoV maturation and facilitating second-round infection. Taken together, this study systematically revaluated and emphasized an essential role of trypsin in PDCoV infection, providing mechanistic insights into the productive infection of PDCoV in porcine-derived cells.

{"title":"Revisiting the roles of trypsin in the productive infection of porcine deltacoronavirus in porcine-derived cells","authors":"Wenwen Xiao , Zhuang Li , Chaoqun Chen , Yuting Shi , Puxian Fang , Shaobo Xiao , Liurong Fang","doi":"10.1016/j.virol.2025.110453","DOIUrl":"10.1016/j.virol.2025.110453","url":null,"abstract":"<div><div>Porcine deltacoronavirus (PDCoV) is an emerging enteric coronavirus with the potential for interspecies transmission. Trypsin has been shown to play a positive role in the isolation and multiplication of PDCoV in vitro, however, the functions of trypsin during PDCoV replication cycle remain controversial. In this study, we revisited the roles of trypsin for PDCoV infection by utilizing two kinds of PDCoV, PDCoV<sup>T+</sup> and PDCoV<sup>T−</sup>, which were prepared in the presence or absence of trypsin, respectively. We found that PDCoV<sup>T+</sup> was able to continuously proliferate in the medium containing trypsin, achieving a higher titer as the infection progress in LLC-PK1 and other tested porcine-derived cells. However, its replication was only transiently improved at 12 hours post-infection, and lower viral titers were observed under trypsin-free culture conditions. Furthermore, the trypsin-mediated enhancement of viral replication could be inhibited by trypsin inhibitor SBTI, suggesting that the second-round viral reproduction of PDCoV<sup>T</sup><sup>+</sup> might be impeded without trypsin. We further investigated the replication dynamics of PDCoV<sup>T−</sup> in LLC-PK1 cells in the presence or absence of trypsin. The results indicated that PDCoV<sup>T−</sup> generated lower viral titers under trypsin-free culture conditions, while the addition of trypsin reverted the infectivity of PDCoV<sup>T−</sup>. Additionally, we demonstrated that trypsin cleaved the PDCoV spike protein, activating viral attachment and internalization. Moreover, trypsin promoted viral replication and release, accelerating PDCoV maturation and facilitating second-round infection. Taken together, this study systematically revaluated and emphasized an essential role of trypsin in PDCoV infection, providing mechanistic insights into the productive infection of PDCoV in porcine-derived cells.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"604 ","pages":"Article 110453"},"PeriodicalIF":2.8,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143420298","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}

引用次数: 0

Complete genomic analysis of tomato chlorosis virus isolates from Sichuan and Chongqing, China

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-11 DOI: 10.1016/j.virol.2025.110447

Xiaolong Yang, Jie Li, Mingrui Gao, Puxin Huang, Mingjun Li, Gentu Wu, Ling Qing

Tomato chlorosis virus (ToCV) has recently emerged as a significant threat to crop production in China. In this study, we collected tomato leaf samples displaying symptoms of interveinal chlorosis, curling, and yellowing from Sichuan Province and Chongqing City. Reverse transcription polymerase chain reaction (RT-PCR) analysis confirmed that 3 out of 10 samples from Sichuan and 9 out of 24 samples from Chongqing were infected with ToCV. Complete genomic sequences of ToCV were assembled from the cDNA of two infected leaves (SC744 and CQ630), which were deposited in GenBank under accession numbers CQ630: PQ153240 and PQ153241 and SC744: PQ261056 and PQ261057. Phylogenetic analysis indicated that ToCV isolates from the Sichuan Province and Chongqing City were closely related to the ToCV-BJ isolate. The sequence identity matrix shows that p22 has the highest variability at 94.3%–98.4%. p6 is the most conserved, with identities between 98% and 100%. In RNA2, p4 varies from 93.9% to 100%, while Hsp70 remains highly conserved at 97.9%–99%. This study provides the first report on the complete genomic sequences of ToCV from Sichuan Province and Chongqing City, enhancing our understanding of ToCV distribution and genetic diversity in China.

{"title":"Complete genomic analysis of tomato chlorosis virus isolates from Sichuan and Chongqing, China","authors":"Xiaolong Yang, Jie Li, Mingrui Gao, Puxin Huang, Mingjun Li, Gentu Wu, Ling Qing","doi":"10.1016/j.virol.2025.110447","DOIUrl":"10.1016/j.virol.2025.110447","url":null,"abstract":"<div><div>Tomato chlorosis virus (ToCV) has recently emerged as a significant threat to crop production in China. In this study, we collected tomato leaf samples displaying symptoms of interveinal chlorosis, curling, and yellowing from Sichuan Province and Chongqing City. Reverse transcription polymerase chain reaction (RT-PCR) analysis confirmed that 3 out of 10 samples from Sichuan and 9 out of 24 samples from Chongqing were infected with ToCV. Complete genomic sequences of ToCV were assembled from the cDNA of two infected leaves (SC744 and CQ630), which were deposited in GenBank under accession numbers CQ630: PQ153240 and PQ153241 and SC744: PQ261056 and PQ261057. Phylogenetic analysis indicated that ToCV isolates from the Sichuan Province and Chongqing City were closely related to the ToCV-BJ isolate. The sequence identity matrix shows that p22 has the highest variability at 94.3%–98.4%. p6 is the most conserved, with identities between 98% and 100%. In RNA2, p4 varies from 93.9% to 100%, while Hsp70 remains highly conserved at 97.9%–99%. This study provides the first report on the complete genomic sequences of ToCV from Sichuan Province and Chongqing City, enhancing our understanding of ToCV distribution and genetic diversity in China.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"604 ","pages":"Article 110447"},"PeriodicalIF":2.8,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143420299","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}

引用次数: 0

Revolutionizing viral resistance strategies in rice: Evolution from RNAi to precision genome editing

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-10 DOI: 10.1016/j.virol.2025.110449

Gaurav Kumar, Indranil Dasgupta

Rice viruses are a major threat to global food security, causing significant yield losses in key rice growing regions. RNA interference (RNAi) has been crucial in engineering viral resistance in rice by silencing essential viral genes. However, the advent of genome editing, especially CRISPR/Cas, has transformed this field by allowing precise alterations of viral susceptibility genes, offering more durable and targeted resistance. This review examines the advances in RNAi strategies and the shift toward CRISPR/Cas technologies, highlighting how genome editing addresses RNAi's limitations, such as broader viral strain coverage and stronger resistance. These tools, integrated with advanced breeding methods, promise to develop rice varieties with durable, broad-spectrum virus resistance, contributing to sustainable rice production and food security.

引用次数: 0

ATF6-mediated mild ER stress inhibits HBV transcription and replication, which is dependent on mTOR activation

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-06 DOI: 10.1016/j.virol.2025.110448

Lin Lu , Ying Feng , Yucai Geng , Zhixiang Liu , Yan Wu , Chen Cai , Ji Zhang , Xingda Huang , Tongchun Xue , Bo Gao

Chronic hepatitis B (CHB) remains a serious global health problem. In our previous investigation, HBV was found to activate a mild ER stress, which facilitated the establishment of persistent HBV infection. However, the role of ER stress manipulation in HBV replication and its underlying mechanisms remain still unclear. Our data showed that mild ER stress inhibited HBV transcription and replication, while severe ER stress enhanced them. Mechanistically, in contrary to the effect on HBV replication, mild ER stress activated whereas severe ER stress inhibited mTOR signaling in HBV-infected cells. Further, mTOR signaling was revealed to be critical for mild ER stress-mediated HBV inhibition. Furthermore, ATF6 but not PERK or IRE1α was found to be involved in mild ER stress-mediated mTOR and the following HBV inhibition. Moreover, ATF6, per se, could inhibit HBV transcription and replication via activating mTOR signaling. Together, ATF6-mediated mild ER stress inhibited HBV transcription and replication through mTOR activation, which might present as an important therapeutic target for CHB patients.

{"title":"ATF6-mediated mild ER stress inhibits HBV transcription and replication, which is dependent on mTOR activation","authors":"Lin Lu , Ying Feng , Yucai Geng , Zhixiang Liu , Yan Wu , Chen Cai , Ji Zhang , Xingda Huang , Tongchun Xue , Bo Gao","doi":"10.1016/j.virol.2025.110448","DOIUrl":"10.1016/j.virol.2025.110448","url":null,"abstract":"<div><div>Chronic hepatitis B (CHB) remains a serious global health problem. In our previous investigation, HBV was found to activate a mild ER stress, which facilitated the establishment of persistent HBV infection. However, the role of ER stress manipulation in HBV replication and its underlying mechanisms remain still unclear. Our data showed that mild ER stress inhibited HBV transcription and replication, while severe ER stress enhanced them. Mechanistically, in contrary to the effect on HBV replication, mild ER stress activated whereas severe ER stress inhibited mTOR signaling in HBV-infected cells. Further, mTOR signaling was revealed to be critical for mild ER stress-mediated HBV inhibition. Furthermore, ATF6 but not PERK or IRE1α was found to be involved in mild ER stress-mediated mTOR and the following HBV inhibition. Moreover, ATF6, <em>per se</em>, could inhibit HBV transcription and replication via activating mTOR signaling. Together, ATF6-mediated mild ER stress inhibited HBV transcription and replication through mTOR activation, which might present as an important therapeutic target for CHB patients.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"604 ","pages":"Article 110448"},"PeriodicalIF":2.8,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143420297","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}

引用次数: 0

Intraspecific SARS-CoV-2 Delta variant transmission among red fox (Vulpes vulpes) and striped skunk (Mephitis mephitis)

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-03 DOI: 10.1016/j.virol.2025.110446

Stephanie M. Porter , Airn E. Hartwig , Marissa Quilici , Angela M. Bosco-Lauth , J. Jeffrey Root

This study assessed the susceptibility of red fox (Vulpes vulpes) and striped skunks (Mephitis mephitis) to the Delta variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Upon finding that both species were susceptible to infection and shed infectious virus, transmission potential was assessed by exposing naïve animals to infected members of their species indirectly (red fox) or directly (striped skunks). Red fox transmitted virus to conspecifics, while contact striped skunks did not become infected with the virus or develop a detectable humoral response. This suggests that red fox could contribute to the maintenance of SARS-CoV-2, while striped skunks are unlikely to do so.

引用次数: 0

Human viruses: An ever-increasing list

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-02 DOI: 10.1016/j.virol.2025.110445

Mei He, Cheng-Qiang He, Nai-Zheng Ding

It has been 124 years since yellow fever was demonstrated to be caused by a ‘filterable agent’. While long-standing viral diseases, with the exception of smallpox, continue to be endemic, new ones have been emerging intermittently, primarily from a substantial zoonotic reservoir, leading to significant socioeconomic consequences. Currently, we are contending not only with the ongoing COVID-19 pandemic but with surges of various other viral infections. Recent metagenomic analyses have revealed a variety of novel human viruses whose health implications remain largely unclear. The following questions arise: How many human viruses have been identified? Which of these viruses are etiological agents of human diseases? This review aims to address the two inquiries and highlight the viruses that pose significant public health threats as outlined in the World Health Organization’s Fact Sheets. Importantly, many human viruses are preventable; appropriate precautions can be implemented to mitigate the risk of infection.

引用次数: 0

Detection of African swine fever virus antibodies using p11.5 and p14.5 protein-based indirect ELISA 基于p11.5和p14.5蛋白的间接ELISA检测非洲猪瘟病毒抗体。

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-01 DOI: 10.1016/j.virol.2024.110335

Aiping Wang , Hehe Zhao , Hongliang Liu , Yumei Chen , Jingming Zhou , Xifang Zhu , Chao Liang , Peiyang Ding , Enping Liu , Gaiping Zhang

African swine fever (ASF), which is caused by the African swine fever virus (ASFV). There are currently no effective vaccines or therapies for ASF; rapid diagnosis has become an important way to control the disease. The p11.5 and p14.5 proteins are structural proteins of ASFV that have immunogenicity and potential as diagnostic antigens. In this study, p11.5 and p14.5 proteins were used as diagnostic antigens in an indirect ELISA examine to detect ASFV antibodies. The p11.5 and p14.5-iELISA methods showed no cross-reaction with other swine virus positive sera. The lowest detection limit of positive serum was 1: 6400, and the coefficients of variation (CV) of intra - and inter-assay were less than 7%. By comparing 71 serum samples, the coincidence rate of p11.5 and p14.5-iELISA with commercial ELISA kits was 91.55%. Taken together, these results showed that there is a reliable method of detecting ASFV antibodies in clinical pig serum.

非洲猪瘟（ASF）是由非洲猪瘟病毒（ASFV）引起的。目前，非洲猪瘟还没有有效的疫苗或疗法；快速诊断已成为控制该疾病的重要方法。p11.5 和 p14.5 蛋白是非洲猪瘟病毒的结构蛋白，具有免疫原性和作为诊断抗原的潜力。在这项研究中，p11.5 和 p14.5 蛋白被用作间接 ELISA 检测 ASFV 抗体的诊断抗原。p11.5 和 p14.5-iELISA 方法与其他猪病毒阳性血清无交叉反应。阳性血清的最低检测限为 1:6400，测定内和测定间的变异系数（CV）均小于 7%。通过比较 71 份血清样本，p11.5 和 p14.5-iELISA 与商业 ELISA 试剂盒的吻合率为 91.55%。综上所述，这些结果表明有一种可靠的方法可以检测临床猪血清中的 ASFV 抗体。

引用次数: 0

Antibody responses in omicron BF.7-infected patients vaccinated with inactivated SARS-CoV-2 灭活SARS-CoV-2疫苗接种组粒bf .7感染患者的抗体应答

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-01 DOI: 10.1016/j.virol.2025.110404

Yabo Zhao , Weili Li , Liming Xu , Zhanling Tao , Junhe Zhang

Objective

Our study aimed to investigate antibody responses in omicron BF.7-infected patients after being vaccinated with inactivated SARS-CoV-2.

Methods

Blood serum samples were collected every 2–7 d, 1 w before infection, during the acute infection period and recovery period, and every month after recovery to detect IgG, IgM, IgA, neutralizing antibodies, and neutralizing antibodies against different omicrons in the acute phase.

Results

The levels of IgG, IgA and neutralizing antibodies increased sharply at 6–7 d after infection, and the levels of IgG and neutralizing antibodies peaked rapidly within 1–2 w, lasting for 3–4 w, and the antibody levels gradually decreased at 4–8 w after infection. The level of neutralizing antibodies against the wild-type strain SARS-CoV-2 was much greater than that against the variant strains during the acute infection period.

Conclusion

After being infected with omicron BF.7, the IgG and neutralizing antibodies in inactivated SARS-CoV-2-vaccinated individuals increase sharply 6–7 days after infection, which is earlier than those in the initial WT SARS-CoV-2 infection, and the peak time within 1–2 w is shorter. The levels of IgG, neutralizing antibody, and IgA antibodies are high, and the levels of IgM antibodies are low; however, the neutralizing antibodies are mainly against the wild-type strain of SARS-CoV-2.

目的：观察组粒bf .7感染患者接种灭活SARS-CoV-2后的抗体反应。方法：每2-7 d、感染前1 w、急性感染期及恢复期、恢复后每个月采集一次血清，检测急性期IgG、IgM、IgA、中和抗体及不同组粒的中和抗体。结果：IgG、IgA及中和抗体水平在感染后6 ~ 7 d急剧升高，IgG及中和抗体水平在感染后1 ~ 2 w内迅速达到峰值，持续3 ~ 4 w，抗体水平在感染后4 ~ 8 w逐渐下降。在急性感染期，对野生型SARS-CoV-2的中和抗体水平远高于变异株。结论：灭活SARS-CoV-2疫苗接种个体感染组粒BF.7后，其IgG和中和抗体在感染后6-7 d内急剧升高，较WT型SARS-CoV-2初始感染时较早，且高峰时间在1-2 w内较短。IgG、中和抗体、IgA抗体水平高，IgM抗体水平低；然而，中和抗体主要针对SARS-CoV-2野生型菌株。

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引用次数: 0

The battle between infectious bronchitis virus and innate immunity: A mini review 传染性支气管炎病毒和先天免疫之间的斗争：一个小回顾。

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-01 DOI: 10.1016/j.virol.2024.110321

Hao Dong , Shengkui Xu , Peng Li , Wenke Ruan

Infectious bronchitis virus (IBV) is the causative agent of infectious bronchitis (IB), leading to acute or persistent infections in poultry. IBV triggers innate immune response, and the production of interferon (IFN) varies depending on the viral strains and host cell types. To evade the host immune system, IBV has developed numerous immune escape strategies. These include hijacking host proteins, modulating protein synthesis, antagonizing IFN production, promoting autophagosome formation and expansion, manipulating apoptosis, blocking antigen presentation, stabilizing viral mRNA, and inhibiting stress granule (SG) formation. The ongoing interaction between IBV and the host immune system reflects a dynamic battle, as the virus employs various tactics to ensure its replication. Understanding these pathogenic mechanisms of IBV is crucial for developing effective control measures.

传染性支气管炎病毒（IBV）是传染性支气管炎（IB）的病原体，可导致家禽急性或持续性感染。IBV触发先天免疫反应，干扰素（IFN）的产生取决于病毒株和宿主细胞类型。为了逃避宿主免疫系统，IBV发展了许多免疫逃逸策略。这些包括劫持宿主蛋白、调节蛋白质合成、拮抗IFN的产生、促进自噬体的形成和扩张、操纵细胞凋亡、阻断抗原呈递、稳定病毒mRNA和抑制应激颗粒（SG）的形成。IBV与宿主免疫系统之间持续的相互作用反映了一场动态的战斗，因为病毒采用各种策略来确保其复制。了解IBV的这些致病机制对于制定有效的控制措施至关重要。

引用次数: 0

Early evolution of mumps-HCV chimeric viruses in Vero cells induces loss of HCV gene expression and promotes accumulation of substitutions uncharacteristic of mumps strains 在Vero细胞中，腮腺炎-丙型肝炎病毒嵌合病毒的早期进化诱导了丙型肝炎病毒基因表达的缺失，并促进了非腮腺炎毒株特征的取代的积累。

IF 2.8 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-02-01 DOI: 10.1016/j.virol.2024.110379

Dorotea Pali , Dubravko Forčić , Maja Jagušić , Tanja Košutić Gulija , Mirna Jurković , Marko Babić , Daniela Kalafatovic , Jelena Ivančić-Jelečki

引用次数: 0

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