Journal of Virology最新文献

{"title":"Deubiquitinase USP8 inhibits HIV-1 replication by suppressing Tat-mediated viral transactivation.","authors":"Jinsong Yuan, Weijing Yang, Mingxiu Xu, Haobo Hu, Shuhan Yang, Xiaodong Sun, Wenyan Zhang, Chen Huan","doi":"10.1128/jvi.00598-26","DOIUrl":"10.1128/jvi.00598-26","url":null,"abstract":"<p><p>Ubiquitin-specific proteases (USPs) modulate diverse virus-host interaction events through deubiquitination; thus, investigating their functional roles and molecular targets facilitates the interpretation of viral pathogenic mechanisms and informs the design of anti-virus strategies. The deubiquitinase USP8 has been reported to antagonize HIV-1 replication by reversing Vif-mediated ubiquitination of the host restriction factor A3G and shielding it from proteasomal degradation. Here, we identify an additional A3G-independent anti-HIV mechanism of USP8. In A3G-negative cells, USP8 exerts a potent inhibitory effect on HIV-1 replication by significantly suppressing viral transcription. Mechanistically, USP8 interacts with the HIV-1 trans-activator Tat through its C-terminal domain and mediates the removal of Tat's polyubiquitination chains in a manner dependent on USP8 catalytic activity. This disrupts the association between Tat and the HIV-1 LTR transactivation response (TAR) element, thus impairing Tat-mediated viral transactivation. Furthermore, given the essential role of Tat in driving viral gene expression, silencing of USP8 triggers latent HIV-1 reactivation. Collectively, our results, along with previous work, reveal that USP8 acts as a compelling anti-HIV factor that, by regulating both host and viral proteins, ultimately suppresses HIV-1 replication. These results deepen our understanding of USP8-mediated HIV-1 restriction and provide insights for the development of novel anti-HIV therapeutic strategies.IMPORTANCEUbiquitination modification contributes a non-negligible part to the regulation of HIV-1 replication, and the interpretation of key factors involved in this process has significantly benefited our understanding of the virus-host interplay. USP8 is a ubiquitin-specific protease that was recently identified to restrict HIV-1 replication by stabilizing the host restriction factor A3G through reversing the Vif-mediated ubiquitination of A3G. Here, we demonstrate an additional A3G-independent anti-HIV mechanism of USP8. In A3G-negative cells, USP8 directly interacts with and deubiquitinates Tat to cleave its K63-linked ubiquitination, thereby suppressing Tat-mediated HIV-1 trans-activation. The suppression of Tat-driven viral transcription induced by USP8 requires the enzymatic activity of USP8 and is effective in different HIV-1 tropic strains. In addition, the fact that Tat is a novel substrate of USP8 is validated both in cells and by <i>in vitro</i> deubiquitination assays. These findings illustrate the dual regulatory roles of USP8 on both host antiviral factors and viral proteins, expanding our understanding of host-mediated HIV-1 restriction.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0059826"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149069893","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"From viral infection to emission: host barriers shaping airborne transmission.","authors":"Patricia Romanos Martinez, Maximilian Rohde, Mohammad Yeasin, Saikat Basu, Christian Sieben, Julia R Port","doi":"10.1128/jvi.00753-26","DOIUrl":"10.1128/jvi.00753-26","url":null,"abstract":"<p><p>Coughing, speaking, and even quiet breathing can release virus-laden particles into the air, triggering transmission events that spread through households and communities. Understanding why some respiratory viruses transmit so efficiently, and under which biological or environmental conditions this occurs, is central to anticipating future outbreaks. In this review, we outline the biological and physical factors that shape respiratory virus transmission, highlighting how the site of replication, mechanisms of droplet generation, and environmental conditions influence airborne spread. We examine how viral traits such as receptor usage, tissue tropism, and innate immune evasion intersect with airway physiology to determine the size, composition, and infectiousness of exhaled particles. We lastly draw attention to key gaps in current knowledge, particularly regarding the host and environmental factors that govern both the release and uptake of virions. Filling these gaps will allow us to more accurately assess transmission risk and implement targeted approaches to reduce spread.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0075326"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149069896","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"UFMylation of SHBs increases protein stability by antagonizing ubiquitination and promotes virion production.","authors":"Shuxiang Wu, Yilin Huang, Jing Xu, Jie Li, Xiaohuang Lin, Xinjian Lin, Xu Lin","doi":"10.1128/jvi.01034-26","DOIUrl":"10.1128/jvi.01034-26","url":null,"abstract":"<p><p>Post-translational modifications orchestrate virus-host interactions, yet their roles in the hepatitis B virus (HBV) life cycle remain incompletely defined. The small hepatitis B surface antigen (SHBs)-the most abundant HBV protein-drives immune evasion, pathogenesis, and virion/subviral particle (SVP) production. Here, we identify UFL1, the E3 ligase of UFMylation, as a host factor that modifies and stabilizes SHBs by antagonizing its ubiquitination to potentiate HBV output and SHBs-driven pro-oncogenic activities. Using affinity purification-mass spectrometry, we discovered UFL1 as a SHBs interactor and validated the interaction by co-immunoprecipitation, confocal co-localization, and GST pull-down assays. <i>In vivo</i> and <i>in vitro</i> UFMylation assays demonstrated that UFL1 catalyzes SHBs' UFMylation. Site-directed mutagenesis mapped the modification to K122; the K122R mutant failed to undergo UFMylation. Cycloheximide chase and ubiquitination assays showed that UFMylation at K122 stabilizes SHBs by antagonizing TRIM21-mediated K48-linked polyubiquitination and proteasomal degradation. Functionally, UFL1 enhanced SHBs-driven hepatocellular carcinoma cell migration and endothelial tube formation, whereas UFL1 knockdown produced opposite effects. In HBV-producing cells, UFL1 increased intracellular and extracellular HBsAg and HBV virions, with no effects on HBV RNA, indicating post-translational control of HBsAg and virion/SVP production. These findings establish SHBs as, to our knowledge, the first HBV protein substrate of UFMylation and reveal a UFL1-SHBs axis that promotes SHBs stability, pathogenic functions, and HBV particle output. Targeting UFMylation or the UFL1-SHBs interface offers a host-directed strategy to reduce HBsAg and virion production and may inform therapeutic approaches for HBV infection and HBV-related liver disease.</p><p><strong>Importance: </strong>Chronic infection with hepatitis B virus (HBV) remains a major global health problem, in part because current treatments rarely reduce the viral surface protein that floods the blood and helps the virus persist. In this study, we asked how the most abundant viral surface protein-SHBs-is controlled inside cells. We discovered that UFL1 attaches a small \"molecular tag\" as UFM1 to this viral protein, and this tag protects the SHBs from the cell's disposal system. As a result, the increased SHBs promotes behaviors linked to liver disease progression, and cells release more virion/SVPs. When we prevented this tagging event or changed a single amino acid on the SHBs so it could not be tagged, the protein became unstable and was degraded, and virus output dropped. Thus, to our knowledge, SHBs is the first identified HBV protein substrate of UFMylation, suggesting targeting the UFL1-SHBs interface may reduce HBsAg and virion production and inform HBV therapy.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0103426"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149017191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PRRSV Nsp9 hijacks BNIP3-mediated autophagy to provide membrane platforms for viral replication complexes.","authors":"Heyou Yi, Shaojun Wang, Guijie Guo","doi":"10.1128/jvi.01314-26","DOIUrl":"10.1128/jvi.01314-26","url":null,"abstract":"<p><p>Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen threatening the global swine industry. The viral nonstructural protein 9 (Nsp9) harbors RNA-dependent RNA polymerase activity and acts as a core component of the viral replication complex, yet its interplay with host factors remains incompletely understood. In this study, we identified the mitochondrial protein BCL2 interacting protein 3 (BNIP3) as a specific binding partner of Nsp9. This interaction is mediated by the transmembrane (TM) domain of BNIP3 and the N-terminal 1-449 region of Nsp9. Functional assays demonstrated that BNIP3 facilitates PRRSV replication via its TM domain and LC3-interacting region (LIR). Mechanistically, BNIP3 recruits Nsp9 to mitochondria, where Nsp9 blocks autophagic flux and triggers incomplete mitophagy. This process leads to the accumulation of BNIP3-associated autophagic membranes, which serve as platforms for viral replication complex assembly. Moreover, simultaneous knockdown of BNIP3 and ATG7 results in further suppression of viral replication compared to knockdown of either gene alone. Rescue experiments showed that BNIP3 overexpression partially restored viral replication in ATG7-deficient cells, while ATG7 overexpression failed to rescue the replication defect caused by BNIP3 knockdown, indicating that BNIP3 possesses ATG7-independent platform functions. Collectively, our findings reveal that PRRSV hijacks BNIP3 to reprogram autophagy and generate membrane platforms for replication, which may serve as a promising target for antiviral development.IMPORTANCEPorcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically significant viral pathogens affecting the global swine industry, yet the specific host factors that support viral replication remain incompletely defined. In this study, we identify BNIP3, a known mitochondrial protein, as a previously unrecognized interactor of the PRRSV RNA-dependent RNA polymerase Nsp9. We demonstrate that PRRSV Nsp9 exploits BNIP3 to block autophagic flux and create a specialized membrane microenvironment that serves as a membrane platform for viral replication complex assembly. Our findings reveal a mechanism by which PRRSV, a positive-sense RNA virus, utilizes a mitophagy receptor to generate replication-permissive membrane platforms, which advances our understanding of how viruses manipulate autophagy. The identification of BNIP3 as a critical host factor that promotes viral replication and the elucidation of its membrane platform function open new avenues for understanding PRRSV replication and may inform the development of novel antiviral strategies.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0131426"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149039568","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expanded protocadherin-1 usage reveals a broader hantavirus entry landscape.","authors":"Cierra Word, Nahomi Guerra-Pilaquinga, Ezgi Kasikci, Lohit Khera, Ramandeep Kaur, Upendra P Lambe, Alexis Ortega, Maria Eugenia Dieterle, Kartik Chandran, Rohit K Jangra","doi":"10.1128/jvi.00994-26","DOIUrl":"10.1128/jvi.00994-26","url":null,"abstract":"<p><p>Mammalian hantaviruses are RNA viruses that cause hantavirus cardiopulmonary syndrome in the Americas and hemorrhagic fever with renal syndrome in Eurasia. The cellular entry mechanisms of most hantaviruses remain poorly defined. To examine entry by phylogenetically distinct hantaviruses identified across Africa, Eurasia, and the Americas, we generated replication-competent recombinant vesicular stomatitis viruses (rVSVs) bearing Gn/Gc proteins from Necoclí, Sangassou, Thottapalayam, Kenkeme, Nova, Oxbow, and Tula viruses. All these Gn/Gc proteins except Kenkeme supported infection of primary human endothelial cells, indicating that endothelial cell entry is permissive for a broader range of hantaviruses than previously appreciated. Except for rVSV-Kenkeme, these rVSVs did not acquire additional mutations beyond pre-engineered rescue-enhancing changes during rescue and passaging. Genetic studies in human cells lacking protocadherin-1 (PCDH1) showed that Necoclí, Tula, and Nova viruses use PCDH1 for efficient infection, although the Nova phenotype was weaker. These three Gn/Gc proteins bound soluble PCDH1 with different apparent avidities, and infection by the corresponding rVSVs was inhibited by soluble PCDH1; Necoclí and Tula, but not Nova, were also blocked by a PCDH1-targeting monoclonal antibody. Authentic Tula virus infection was similarly reduced in PCDH1 knockout endothelial cells. Finally, the broadly reactive anti-Gn/Gc human monoclonal antibody ADI-42898 efficiently neutralized Necoclí, Nova, Sangassou, and Kenkeme rVSVs, but showed weak or undetectable activity against Oxbow, Tula, and Thottapalayam rVSVs. Together, these findings expand the range of hantavirus glycoproteins capable of mediating infection of human endothelial cells, broaden the phylogenetic scope of PCDH1-dependent entry, and identify receptor-targeted and viral glycoprotein-targeted strategies with differential activity.IMPORTANCEMany newly discovered hantaviruses are known only from sequence data, leaving their ability to enter human cells and their receptor usage unresolved. Using a BSL2-compatible recombinant vesicular stomatitis virus (rVSV) system, we show that glycoproteins from several divergent hantaviruses can mediate infection of primary human endothelial cells, indicating that endothelial cell entry is permissive for a broader range of hantaviruses than previously appreciated. We also show that protocadherin-1 (PCDH1), previously linked mainly to New World hantaviruses, is used by Necoclí, Tula, and Nova viruses but not universally across the panel, revealing broader but heterogeneous receptor usage. An authentic Tula virus experiment supports this conclusion beyond the surrogate system. Finally, a broadly reactive anti-Gn/Gc antibody neutralizes several, but not all, of these viruses, highlighting both the promise and the limits of broadly protective countermeasures and the utility of these rVSVs for evaluating entry inhibitors.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0099426"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149075073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Virus-specific T cells and neutralizing antibodies are independent correlates of long-term protective immunity against hepatitis C virus.","authors":"Elsa Gomez-Escobar, Asiyah Siddique, Omar Khedr, Nathalie Bédard, Alizée Fieffé-Bédard, Nourtan F Abdeltawab, Nicol Flores, Mohamed S Abdel-Hakeem, Mohamed Eisa, Geng Zang, John Lok Man Law, Juthika Kundu, Michael Houghton, Julie Bruneau, Naglaa H Shoukry","doi":"10.1128/jvi.00849-26","DOIUrl":"10.1128/jvi.00849-26","url":null,"abstract":"<p><p>Virus-specific T cells and broadly neutralizing antibodies (NAbs) are associated with spontaneous clearance of acute primary hepatitis C virus (HCV) infection and reinfection. Following spontaneous clearance, HCV-specific memory T cells are long-lived, but NAbs are less durable. How these two arms contribute to long-term protective immunity upon re-exposure is unknown. Herein, we compared the magnitude and breadth of memory HCV-specific T cells and NAbs in a cohort of HCV-spontaneously resolved people who inject drugs with high-risk exposure to the virus but who remained free of observed reinfections (FOR, <i>n</i> = 34) or who got reinfected (<i>n</i> = 22). Among FOR, 73.5% (<i>n</i> = 25) exhibited high frequencies of gamma interferon (IFN-γ)-producing T cells, while 23.5% (<i>n</i> = 8) showed high neutralization breadth (>25% neutralization of HCV pseudoparticles [HCVpp]) and/or potency (geometric mean neutralization >50% against seven HCVpp). Notably, some subjects had detectable NAbs several years after clearance. Multivariate random forest analysis demonstrated that the frequency of IFN-γ-producing T cells and the NAb response against only one difficult-to-neutralize HCVpp (1b58) were significant predictors of protection against reinfection (<i>P</i> < 0.05). Longitudinal analysis of the immune response in subjects with multiple episodes of infection showed that strong and broad HCV-specific T cell responses were associated with spontaneously cleared episodes. In contrast, NAbs did not protect from chronicity upon re-exposure in the absence of memory T cells, or if they were unable to recognize the infecting virus due to imprinting by strains of the previous episodes. In conclusion, T cell responses and NAbs targeting specific isolates are independent predictors of long-term protective immunity against HCV.</p><p><strong>Importance: </strong>Hepatitis C virus (HCV) is a blood-borne virus that disproportionately affects people who inject drugs. Most HCV infections become persistent, leading to liver fibrosis and cancer. Although effective antiviral therapies are available, around 50 million people remain persistently infected with HCV and can transmit the virus. Currently, there is no available vaccine to control the spread of HCV. However, around one in four people infected can naturally clear the virus, becoming partially protected upon re-exposure, suggesting it is possible to develop a vaccine that induces similar protection. In this study, we investigated the contribution of T cells and neutralizing antibodies to protection against observed reinfection in a cohort of people who inject drugs and have naturally cleared HCV, yet remain at high-risk exposure. Our findings suggest that strong T cell responses and neutralizing antibodies targeting hard-to-neutralize isolates could predict protective immunity upon reinfection.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0084926"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148920210","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The multi-pronged entry of African swine fever virus: from host factors to therapeutic targets.","authors":"Jiaqi Li, Chuanxia Liu, Weitao Huang, Yibing Wang, Li Huang, Changjiang Weng","doi":"10.1128/jvi.00924-26","DOIUrl":"10.1128/jvi.00924-26","url":null,"abstract":"<p><p>African swine fever (ASF), caused by the African swine fever virus (ASFV), remains one of the most devastating transboundary animal diseases worldwide, with case fatality rates reaching up to 100% in acute infections. Viral entry represents the first essential step in infection and is an ideal target for the development of vaccines and antiviral drugs. Recent studies have revealed that ASFV entry is not mediated by a single receptor or pathway. Instead, it involves multiple viral structural proteins, cellular attachment factors, endocytic routes, and intracellular trafficking regulators, which form a coordinated virus-host interaction network. In this review, we systematically summarize recent advances in ASFV multi-pathway invasion, including viral attachment, internalization, intracellular trafficking, uncoating, and membrane fusion. Additionally, we discuss several unresolved questions and highlight promising antiviral and genetic intervention strategies that target both viral components and host determinants. A comprehensive understanding of ASFV entry will provide the theoretical foundation for developing next-generation vaccines, antivirals, and disease-resistant pig populations.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0092426"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149025285","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rift Valley fever virus virulence factor NSs targets ATF4- and XBP1-driven unfolded protein response.","authors":"Kaori Terasaki, Shinji Makino","doi":"10.1128/jvi.01233-26","DOIUrl":"10.1128/jvi.01233-26","url":null,"abstract":"<p><p>Rift Valley fever virus (RVFV) is a mosquito-borne arbovirus that infects humans and domestic ruminants, significantly impacting public health and the economy in African and Middle Eastern countries. RVFV non-structural protein NSs is a major viral virulence factor and possesses multiple functions to counteract host antiviral responses. One of its functions is to block host transcription by promoting degradation of subunits of the host transcription factor IIH (TFIIH). However, the biological significance of this function in the RVFV life cycle remains unclear. We used RVFV strain MP-12 and its mutant lacking the TFIIH degradation function (NSs-mut virus) to identify host genes targeted by NSs-mediated transcription suppression. We found that NSs-mut virus, but not MP-12, upregulated unfolded protein response (UPR) target genes compared to uninfected cells. Both viruses activated all three ER stress sensors, including PERK and IRE1. However, only MP-12 inhibited the PERK and IRE1 pathways by inhibiting the expression of their respective downstream mediators ATF4 and XBP1. ATF4 expression was inhibited at the transcriptional level, whereas XBP1 expression was inhibited at the post-transcriptional level. ATF4 depletion negatively impacted virus titer and the production of non-plaque-forming particles, resulting in alteration of the particle-to-PFU ratio of the viruses. We also showed that activation of the PERK pathway plays a pro-survival role in NSs-mut virus-infected cells, suggesting the involvement of the PERK pathway in RVFV-induced cytotoxicity. Taken together, our study revealed that RVFV NSs inhibits two out of the three UPR pathways and promotes RVFV-induced cytotoxicity by inhibiting the PERK pathway.IMPORTANCEThe Rift Valley fever virus (RVFV) NSs protein is a major virulence factor with multiple functions, including inhibition of general host transcription. RVFV carrying a mutated NSs that lacks the transcription suppression function is less cytotoxic <i>in vitro</i> and less virulent in mice compared to its parental virus, suggesting that NSs-mediated host transcription suppression is important for RVFV virulence. Experiments using this mutant virus and its parental RVFV revealed that NSs inhibits the expression of unfolded protein response (UPR) mediators ATF4 and XBP1, which act downstream of the PERK and IRE1 pathways, respectively. Our data also showed that activation of PERK promotes cell survival in RVFV infection. These data suggest that the NSs-mediated inhibition of UPR pathways contributes to RVFV pathogenicity.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0123326"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149039552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mutations that positively affect <i>Bandavirus</i> glycoprotein function in VSV-vectored vaccines.","authors":"Raegan J Petch, Philip Hicks, Jonna B Westover, Brian B Gowen, Paul Bates","doi":"10.1128/jvi.01311-26","DOIUrl":"10.1128/jvi.01311-26","url":null,"abstract":"<p><p>Severe fever with thrombocytopenia syndrome virus (SFTSV) and Heartland bandavirus (HRTV) are emerging tick-borne bandaviruses. They have high case fatality rates (10%), and no FDA-approved vaccines exist for disease prevention. SFTSV and HRTV are therefore identified as priority pathogens. A recombinant vesicular stomatitis virus (rVSV) vaccine, which replaces the original VSV glycoprotein with the SFTSV glycoproteins, shows early promise for SFTSV as it induces strong immune responses that are protective against lethal challenge. However, rVSV-SFTSV is highly attenuated in cell culture, which may be due to incompatibility between the assembly sites of SFTSV (the Golgi and endoplasmic reticulum-Golgi intermediate compartment [ERGIC]) and those of VSV (the plasma membrane). In this study, we identify a noncanonical coatomer complex I (COPI)-binding motif found in the cytoplasmic tail of SFTSV glycoproteins and demonstrate that an amino acid substitution in this motif (K1071A) inhibits binding to COPI. This mutation results in increased surface expression of SFTSV glycoproteins, improved incorporation onto VSV virions, and enhanced replication of rVSV-SFTSV <i>in vitro</i>. A mutation at a homologous site (K1074A) in HRTV yields similar results, and rVSV-HRTV K1074A exhibits increased replication <i>in vitro</i> and <i>in vivo</i>. We show that vaccination with rVSV-HRTV K1074A results in improved induction of neutralizing antibody responses in immunocompetent C57BL/6 mice, and neutralizing antibodies elicited by vaccination are protective when administered to severely immunocompromised mice via passive transfer. Overall, our study identifies a mutation that improves the efficacy of the rVSV-SFTSV vaccine candidate and introduces the first vaccine candidate directly addressing HRTV infections.IMPORTANCESevere fever with thrombocytopenia syndrome virus (SFTSV) and Heartland bandavirus (HRTV) are emerging tick-borne viruses with high fatality rates. FDA-approved vaccines and antiviral drugs are unavailable but critically needed. We identify an important mutation in the SFTSV glycoprotein that disrupts a previously unreported COPI-binding site. The mutation improves the efficacy of the previously described recombinant vesicular stomatitis virus vaccine candidate for SFTSV (rVSV-SFTSV). We also develop an rVSV-HRTV vaccine and show potent induction of neutralizing antibodies and protection from lethal challenge. This is the first study directly addressing the lack of vaccines specifically targeting HRTV.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0131126"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149039594","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Real-time tracking of vaccinia virus entry using a split luciferase complementation assay.","authors":"Yunliang Zhang, Jaroslav Holly, James Gibbs, Jonathan W Yewdell, Bernard Moss","doi":"10.1128/jvi.01115-26","DOIUrl":"10.1128/jvi.01115-26","url":null,"abstract":"<p><p>A recombinant vaccinia virus (vA4-HiBiT) was constructed with a short, split luciferase HiBiT tag at the C-terminus of the A4 core protein open reading frame. When HeLa or A549 cells stably expressing LgBiT were infected with vA4-HiBiT, the interaction of the split luciferase fragments reconstituted NanoLuc enzyme activity. Upon addition of a membrane-permeable luciferase substrate, a strong luminescence signal indicated core entry into the cytoplasm within 5 min after infection and was proportional to virus multiplicity within the range of 0.5-80 PFU/cell. This direct entry assay was dependent on a functional virus entry-fusion complex, was inhibited by a neutralizing antibody, and was unaffected by an inhibitor of protein synthesis. Entry was enhanced by brief pH 5 treatment of cell-bound virus relative to pH 7 and prevented by pH 3 treatment. Bafilomycin A1, an inhibitor of endosomal acidification, exhibited greater inhibition of entry in A549-LgBiT cells compared to HeLa-LgBiT cells, as did treatment with ammonium chloride, suggesting variations in entry pathways or cell metabolism. Inhibition of entry by latrunculin A and cytochalasin D indicated a role for actin polymerization in both cell lines. Despite the discovery of a conserved furin cleavage consensus sequence in the A16 subunit of the entry-fusion complex, a furin inhibitor did not reduce virus entry unlike results obtained with a human papillomavirus control. These data support the use of this NanoBiT assay for studies of vaccinia virus entry, with application to basic research, antivirals, and neutralizing antibodies.</p><p><strong>Importance: </strong>Assays to measure virus entry into cells mostly rely on post-entry events such as gene expression, cytopathic effects, or virus yields. Although fluorescence and electron microscopy can provide direct analyses, such methods are labor intensive and semiquantitative. Here we describe a rapid and quantitative real-time direct entry assay for vaccinia virus with application to basic research, antivirals, and neutralizing antibodies.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0111526"},"PeriodicalIF":4.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149075242","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}