Journal of Virology最新文献

{"title":"Revisiting mRNA capping in <i>Mononegavirales</i>: a promising, yet underexplored antiviral target.","authors":"Khadijah Khan, Mohammad Ejaz Adil, Monika Antil, Dilip Kumar","doi":"10.1128/jvi.01025-26","DOIUrl":"https://doi.org/10.1128/jvi.01025-26","url":null,"abstract":"<p><p>mRNA capping is a co-transcriptional, nuclear process in eukaryotes that adds a cap onto the 5' end of RNA to confer stability, efficient translation, and nuclear export. Viruses have evolved diverse strategies by mimicking or subverting host pathways to install an mRNA cap for efficient gene expression and evasion of the host innate immune response. The conventional mRNA capping pathway, employed by many RNA and DNA viruses, is mediated by dedicated RNA 5'-triphosphatase, guanylyltransferase, and methyltransferase enzymes. In contrast, many RNA viruses utilize unconventional capping mechanisms that diverge from the canonical pathway, offering unique opportunities to develop virus family-specific antiviral therapeutics. Among negative-sense RNA viruses, members of the order <i>Mononegavirales</i> harbor a unique capping domain, polyribonucleotidyltransferase (PRNTase), embedded within the viral RdRp large (L) protein. Unlike the conventional guanylyltransferase-mediated mechanism, PRNTase catalyzes mRNA capping through the formation of a covalent L-pRNA intermediate rather than the canonical GMP-enzyme intermediate. The HR motif in PRNTase, involved in covalent linking to mRNA, is highly conserved in non-segmented negative-strand RNA viruses. This review focuses on structural and functional insights into <i>Mononegavirales</i> capping, highlighting the conserved motifs and domain architecture that make it a promising therapeutic target.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0102526"},"PeriodicalIF":4.1,"publicationDate":"2026-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149182385","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":"Avian influenza A virus polymerase gene leads to temperature-resistant genome replication.","authors":"Stephanie L Williams, Sebastian M Gygli, Jacob C Bjorgen, Haitian Fan, Munashe Mudiwa, Lex Matthews, Yongli Xiao, Ashley Freeman, Sharon F Legaspi, Li Qi, Jeffery K Taubenberger, Ervin Fodor","doi":"10.1128/jvi.00978-26","DOIUrl":"https://doi.org/10.1128/jvi.00978-26","url":null,"abstract":"<p><p>Influenza A viruses (IAV) pose a persistent pandemic threat. Successful cross-species transmission from waterfowl to mammals requires host adaptation for efficient replication and spread. Avian IAVs are adapted to the temperature of the avian gut, approximately 41°C, while human IAVs replicate at 33°C in the upper respiratory tract. PB1, the catalytic core of the viral heterotrimeric RNA-dependent RNA polymerase (RdRp), is essential for viral RNA synthesis. Three pandemic IAV strains (1918, 1957, and 1968) carried an avian-origin PB1 gene, suggesting a selective advantage. Fever, a critical antiviral response, elevates body temperature to 38°C or higher and can restrict viral replication. We hypothesized that avian PB1 may allow IAV replication at febrile temperatures. Using a mini-replicon system, we demonstrate that a polymerase of a seasonal human H1N1 IAV fails to replicate viral RNA efficiently at 40°C, whereas substitution of the PB1 with that from the 1918, 1957, and 1968 pandemic viruses, or a low-pathogenic avian H2N1 virus, restores RNA synthesis. Chimeric seasonal human H1N1 viruses expressing avian or 1918 PB1 replicate at 40°C in MDCK cells, unlike wild-type H1N1. Activity assays with purified chimeric IAV polymerases further suggest that avian-origin PB1 may intrinsically enhance polymerase activity at elevated temperatures. Together, these data show that avian-origin PB1 enables IAV replication at febrile temperatures, highlighting its role in zoonotic emergence and host adaptation. Surveillance of avian-origin PB1 genes in IAVs circulating in mammals may help identify strains with pandemic potential and inform the development of countermeasures.IMPORTANCEInfluenza A viruses of avian origin remain a persistent threat to human health. The 1918, 1957, and 1968 pandemic influenza viruses each acquired an avian-origin PB1 gene, but the adaptive advantage conferred by these genes remains incompletely understood. Here, we show that avian-origin PB1 genes enhance viral RNA replication by a seasonal human influenza virus polymerase at elevated temperature and enable replication of a human influenza virus at 40°C, a temperature relevant to both avian hosts and human febrile illness. PB1-dependent effects were observed in cell-based polymerase assays, with purified polymerase, and during authentic virus infection. These findings identify temperature-resistant genome replication as a potential adaptive property of avian-origin PB1 and suggest that acquisition of an avian PB1 may help influenza viruses overcome temperature barriers encountered during adaptation to mammalian hosts.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0097826"},"PeriodicalIF":4.1,"publicationDate":"2026-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149182721","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":"Superinfection exclusion strategy of siphophage T5: analysis of the FhuA:Llp complex.","authors":"Séraphine Degroux, Corinne Vivès, Emeline Mestdach, Claudine Darnault, Aline le Roy, Caroline Mas, Loïc Salmon, Torsten Herrmann, Cécile Breyton","doi":"10.1128/jvi.00909-26","DOIUrl":"https://doi.org/10.1128/jvi.00909-26","url":null,"abstract":"<p><p>Superinfection exclusion is a widespread viral mechanism that protects the infected cell from overinfection by other identical or closely related viruses. Shortly after infection by bacteriophage T5, the bacterium <i>Escherichia coli</i> produces Llp, encoded by the newly injected viral genome. Llp is a small lipoprotein targeted to the inner leaflet of the host outer membrane. It binds to FhuA, an outer-membrane iron-ferrichrome transporter, which is the T5 receptor at the <i>E. coli</i> cell surface. The interaction between Llp and FhuA prevents any further bacteriophage binding and infection. Here, we determined the NMR structure of Llp and analyzed the formation of the FhuA:Llp complex using a wide range of techniques and mutants, both <i>in vivo</i> and <i>in vitro</i>. Interaction of Llp with FhuA is governed by a two-step equilibrium with a strong contribution of induced fit: Llp binding requires remodeling of FhuA periplasmic plug surface, enabling the subsequent large conformational reorganization of the extracellular loops. Analysis of FhuA mutants shows the importance of the intertwined interactions between the extracellular loops and the plug in the communication between the periplasmic and the extracellular faces of FhuA.IMPORTANCEBacteriophages (phages) are viruses that infect bacteria. As their enemies, phages play an important role in the dynamics, diversity, evolution, and pathogenicity of bacterial populations, but also represent an alternative to antibiotics in our fight against the rise of multi-resistant pathogenic bacterial strains. Infection is triggered by phage recognition of a receptor at the host surface, inducing phage DNA injection into the bacterial cytoplasm. The host cell is then transformed into a viral factory. To secure this viral factory, the infecting phage develops superinfection exclusion strategies, preventing other phages from interfering. Understanding how phages achieve the \"locking\" of the host will allow us to optimize phage use in phage therapy. Here, we analyze the structure of the receptor-exclusion protein complex using integrated <i>in vivo</i>, biochemical, and biophysical approaches. Our analysis reveals an unusually slow association process, consistent with an induced-fit mechanism, ultimately transmitting information from the bacterial periplasm to its surface.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0090926"},"PeriodicalIF":4.1,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149167220","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":"Marek's disease virus: mechanistic insights into host immune responses and multilayered immune evasion strategies.","authors":"Kexin Gao, Qingqing Yu, Ru Guo, Huijing Sun, Yulu Duan, Yanping Zhang, Yulong Gao, Yongzhen Liu","doi":"10.1128/jvi.00736-26","DOIUrl":"https://doi.org/10.1128/jvi.00736-26","url":null,"abstract":"<p><p>Marek's disease virus (MDV) is a highly contagious, oncogenic alphaherpesvirus that induces profound immunosuppression and T-cell lymphomas in poultry, serving as a prominent paradigm for investigating virus-induced tumorigenesis and pathogen evolution under vaccine-driven selection pressure. Despite half a century of widespread vaccination, the continuous circulation and progressive enhancement of MDV virulence within flocks highlight a critical gap in our understanding of host-virus immunological interplay and co-evolution. This review systematically delineates the dynamic molecular mechanisms underlying the interplay between MDV and the host immune system. We first dissect the innate immune sensing networks, with a particular focus on the cGAS-STING pathway, and elucidate the multidimensional strategies employed by MDV to antagonize these early antiviral defenses. Subsequently, we explore the complexities of the adaptive immune response, revealing the immune evasion mechanisms by which MDV establishes a tumor-permissive microenvironment via the downregulation of MHC-I, induction of T-cell exhaustion, and recruitment of regulatory T cells. Furthermore, we discuss telomere integration as a key mechanism contributing to MDV persistence and highlight the unresolved mechanisms governing reactivation. The purpose of this review is to clarify the key factors that determine the immune protective efficacy of MDV vaccines and to provide insights into the optimization of prevention and control strategies.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0073626"},"PeriodicalIF":4.1,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149170007","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":"HSV-1 induces stable associations between the Golgi and endosomes within the cytoplasmic viral assembly compartment of neuronal cells.","authors":"Dah Hyun Jung, Shaowen White, Richard J Roller","doi":"10.1128/jvi.00999-26","DOIUrl":"https://doi.org/10.1128/jvi.00999-26","url":null,"abstract":"<p><p>Assembly of mature herpesvirus particles is associated with extensive rearrangement of cytoplasmic membranes. Herpes simplex virus type 1 (HSV-1) infection in neuronal cells induces the formation of a distinctive cytoplasmic viral assembly compartment (cVAC), in which rearranged host membranes and viral proteins are clustered at the perinuclear microtubule-organizing center (MTOC). Although both the <i>trans</i>-Golgi network (TGN) and endosomal compartments have been implicated as membrane sources for cytoplasmic envelopment, how secretory and endosomal pathways are reorganized and associated within the cVAC remains unclear. Here, we treated HSV-1-infected cells with nocodazole at a post-entry stage to disperse membrane components of the cVAC and identify microtubule-independent membrane organizations and associations using immunofluorescence imaging. We found that the Golgi apparatus was fragmented during infection but maintained cisternal stacking, as the Golgi markers GM130, GalT, and GRASP65 remained colocalized in both the absence and presence of nocodazole. We further demonstrated that Golgi mini-stacks retained their association with Rab6 during HSV-1 infection and formed microtubule-independent associations with early and recycling endosomes, resulting in the formation of Golgi-endosome super-mini-stacks. Viral capsid, glycoprotein, and tegument proteins were colocalized with Golgi-endosome super-mini-stacks in both the presence and absence of microtubules, suggesting that this membrane complex serves as a site of cytoplasmic envelopment regardless of clustering at the MTOC. Formation of these stable Golgi-endosome associations did not require viral capsids. Neither actin filaments nor acetylated microtubules colocalized with Golgi-endosome super-mini-stacks, suggesting that cytoskeletal elements are not required for this association.IMPORTANCEMany viruses remodel host cytoplasmic membranes to facilitate their assembly. Elucidating the molecular mechanisms underlying this remodeling may reveal novel antiviral targets while also providing insight into fundamental principles of intracellular membrane organization. Infection with an important human pathogen, herpes simplex virus type 1 (HSV-1), induces extensive membrane remodeling that leads to the formation of the cytoplasmic viral assembly compartment (cVAC) in neuronal cells, yet the underlying mechanisms remain poorly understood. Here, we demonstrate that HSV-1 induces stable associations between membranes within the cVAC. Our findings support a model in which HSV-1 generates a dynamic hybrid membrane compartment during infection, potentially reconciling conflicting views on the roles of distinct membrane sources in cytoplasmic assembly.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0099926"},"PeriodicalIF":4.1,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149170093","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":"Syncrip coordinates with hnRNPK to regulate PPV NS1 mRNA splicing and facilitate viral replication.","authors":"Yiyuan Jiang, Qian Du, Xiaoyang Zhang, Liu Yang, Caiyi Chen, Bichen Miao, Yong Huang, Dewen Tong","doi":"10.1128/jvi.00901-26","DOIUrl":"https://doi.org/10.1128/jvi.00901-26","url":null,"abstract":"<p><p>Porcine parvovirus (PPV) relies on alternative splicing of the nonstructural protein 1 (NS1) transcript to generate nonstructural protein 2 (NS2), but the host factors coordinating this process remain incompletely defined. Here, we identified heterogeneous nuclear ribonucleoprotein K (hnRNPK) as a regulator of PPV NS1-to-NS2 mRNA splicing in PK-15 cells. hnRNPK specifically associated with NS1 mRNA, and PPV infection increased hnRNPK expression in a time- and multiplicity-of-infection-dependent manner. hnRNPK overexpression decreased the NS1/NS2 mRNA and protein ratios and increased viral DNA accumulation, whereas hnRNPK knockout produced the opposite effects. hnRNPK and the previously identified splicing regulator synaptotagmin-binding, cytoplasmic RNA-interacting protein (Syncrip) additively promoted NS1-to-NS2 splicing and PPV replication. Double-knockout and reconstitution experiments further showed that co-expression of hnRNPK and Syncrip restored NS2 production and viral replication more effectively than either factor alone. Domain-mapping analyses identified the K-protein interaction region (KI) of hnRNPK and the RNA recognition motif 2 (RRM2) region of Syncrip as the principal regions supporting this activity. hnRNPK associated directly with Syncrip, and Syncrip was required for efficient recruitment of hnRNPK to NS1 mRNA. In addition, hnRNPK associated with serine- and arginine-rich splicing factor 1 (SRSF1) in an RNA-independent manner during PPV infection. These findings support a model in which a Syncrip-hnRNPK complex promotes PPV NS1-to-NS2 alternative splicing and thereby facilitates viral replication, while identifying SRSF1 as a candidate component of the associated host splicing machinery.IMPORTANCEPorcine parvovirus (PPV) causes reproductive failure in swine and relies extensively on host RNA-processing machinery because of its limited coding capacity. Alternative splicing of the PPV nonstructural protein 1 (NS1) transcript generates nonstructural protein 2 (NS2), but the host factors controlling this process remain poorly defined. Here, we identify heterogeneous nuclear ribonucleoprotein K (hnRNPK) as a host regulator of PPV mRNA splicing and replication. hnRNPK acts additively with synaptotagmin-binding, cytoplasmic RNA-interacting protein (Syncrip) to promote NS1-to-NS2 mRNA splicing, increase NS2 production, and support viral DNA replication. Syncrip facilitates the association of hnRNPK with NS1 mRNA, and defined regions within both proteins contribute to this activity. hnRNPK also associates with the cellular splicing factor serine- and arginine-rich splicing factor 1 (SRSF1) during PPV infection. These findings reveal a host RNA-binding protein network that PPV exploits to expand its coding capacity and promote replication.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0090126"},"PeriodicalIF":4.1,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149172233","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":"SERINC5 is the primary target of retroviral antagonists among human SERINC paralogs and requires a tri-cysteine motif for its counteraction by HIV-1 Nef.","authors":"Cinzia Bertelli, Annachiara Rosa, Teresa Vanzo, Irene Crescioli, Hamza Ur Rehman, Ajit Chande, Remigiusz Serwa, Edward W Tate, Peter Cherepanov, Massimo Pizzato","doi":"10.1128/jvi.01088-26","DOIUrl":"https://doi.org/10.1128/jvi.01088-26","url":null,"abstract":"<p><p>The human transmembrane protein SERINC5 incorporates into budding HIV-1 particles and inhibits virus fusion with target cells. Although three other human SERINC paralogs display antiviral activity, their contribution to retroviral restriction and their susceptibility to viral antagonists have not been systematically compared. HIV-1 Nef, Moloney murine leukemia virus (MoMLV) glycoGag, and the equine infectious anemia virus (EIAV) S2 glycoprotein independently evolved mechanisms to counteract SERINC5. Here, we show that all human SERINC paralogs are incorporated into HIV-1 virions despite their differing antiviral activities. However, compared with the other SERINC paralogs, SERINC5 is selectively and efficiently targeted by all retroviral antagonists examined, supporting the notion that SERINC5 has exerted the strongest selective pressure on retroviral evolution. We identified a tri-cysteine motif within intracellular loop 4 (ICL4) as a critical determinant of SERINC5 antagonism by Nef. Disruption of this motif impaired Nef-mediated internalization, exclusion from virions, and full rescue of HIV-1 infectivity by diverse HIV-1 and SIV Nef alleles, but had no effect on antagonism by MoMLV glycoGag or EIAV S2, indicating that these viral proteins recognize distinct molecular features of SERINC5. Transfer of ICL4 to SERINC2 was insufficient to confer Nef sensitivity, demonstrating that the tri-cysteine motif is necessary but not sufficient for antagonism. Finally, we show that this motif is palmitoylated, suggesting that palmitoylation contributes to SERINC5 recognition by Nef. Together, these findings establish SERINC5 as the principal target of retroviral antagonists and demonstrate that its recognition by Nef depends on the broader structural context of SERINC5.</p><p><strong>Importance: </strong>The SERINC family of multipass transmembrane proteins includes host factors capable of restricting retrovirus infectivity. Among the five human paralogs, SERINC5 displays the strongest antiviral activity and is the only member efficiently targeted by all known retroviral antagonists, including HIV-1 Nef, MLV glycoGag, and the EIAV S2. This selectivity indicates that the evolutionary pressure exerted by SERINC proteins on retroviruses has been primarily driven by SERINC5 rather than by its relatives. Through structure-guided mutagenesis, we identified a tri-cysteine motif within intracellular loop 4 as a crucial determinant for SERINC5 downregulation by Nef, providing a molecular explanation for its distinctive susceptibility. These findings establish SERINC5 as the principal target of viral counteraction and suggest that its molecular features have contributed to shaping the adaptation of primate lentiviral Nef proteins to the host.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0108826"},"PeriodicalIF":4.1,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149170716","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":"Temporal dissection of hepatitis C virus and orthoflavivirus infections by continuous luminescence monitoring.","authors":"Masahiko Ito, Xiaoyu Li, Jie Liu, Mami Matsuda, Kwang Su Kim, Toru Takada, Haruyo Aoyagi, Takayuki Hishiki, Fumihiro Kato, Shigeru Tajima, Kodama Watanabe, Mana Watanuki, Kaho Matsumura, Alu Konno, Koichi Watashi, Hideki Aizaki, Masayoshi Fukasawa, Takanobu Kato, Takaji Wakita, Yasumasa Iwatani, Shingo Iwami, Ryosuke Suzuki, Tetsuro Suzuki","doi":"10.1128/jvi.01266-26","DOIUrl":"https://doi.org/10.1128/jvi.01266-26","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Virus infection is a dynamic process consisting of rapidly evolving viral and host events, yet conventional endpoint assays provide only fragmented snapshots of these processes. Here, we established a unified temporal framework for dissecting virus-host interactions by integrating continuous bioluminescence monitoring with molecular and functional analyses. Real-time monitoring of hepatitis C virus (HCV) single-round infectious particles (SRIPs) revealed a biphasic replication profile consisting of an initial transient phase, followed by replication-dependent amplification. This transition was independently validated by similar HCV RNA kinetics observed after infection with HCV SRIPs or fully infectious HCV. Extension of this approach to dengue, Zika, and yellow fever virus SRIPs demonstrated that infection kinetics are determined by virus-specific replication programs and are modulated by the combination of viral replicons/capsids and prM/E proteins. This temporal framework enabled the identification of virus-host interactions that are difficult to resolve by conventional endpoint analyses. Early-phase cytoplasmic proteomics, combined with functional screening, identified the dynein light chain component DYNLRB2 as a previously unrecognized negative regulator of early HCV infection. Continuous monitoring also distinguished phase-specific antiviral activities, revealing distinct kinetic inhibition profiles for daclatasvir, acetylsalicylic acid, and interferon-α. Furthermore, delayed activation of the ER stress sensor ATF6 was first detected 40-47 h after HCV infection, and transcriptomic analysis at reporter-defined time points characterized the accompanying transition in host gene expression. Together, our study establishes continuous temporal monitoring as a general strategy for resolving dynamic virus-host interactions and for uncovering molecular events that remain obscured by conventional endpoint analyses.IMPORTANCEViral infection is a dynamic process in which entry, genome amplification, host-factor requirements, antiviral effects, and cellular responses occur on different timescales. Conventional endpoint assays can identify changes in these processes but provide limited information about when they emerge or transition from one stage to another. Here, we show that continuous monitoring, when combined with direct molecular and functional measurements, can place these otherwise separate observations within a common temporal framework. Using HCV and related orthoflavivirus SRIPs, we resolved early and replication-associated phases, identified host factors with stage-dependent effects, and distinguished profiles of antiviral inhibition. We further showed that ATF6-mediated ER stress activation emerged later than early HCV infection and viral RNA amplification. Thus, continuous temporal monitoring provides a general strategy for resolving dynamic virus-host interactions by defining the temporal relationships among viral processes, pharma","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0126626"},"PeriodicalIF":4.1,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149166397","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":"A multivalent mRNA-lipid nanoparticle vaccine containing eight hemagglutinin antigens elicited broad neutralizing antibody responses and protected against influenza A virus challenge in swine.","authors":"Giovana Ciacci Zanella, Marissa L Vincent, Leo Flores, Ethan K Aljets, Rodrigo C Paiva, Alexey Markin, Blake Inderski, Garima Dwivedi, Drew Weissman, Meghan Wymore Brand, Jefferson J S Santos, Scott E Hensley, Tavis K Anderson, Phillip C Gauger, Amy L Baker","doi":"10.1128/jvi.00848-26","DOIUrl":"https://doi.org/10.1128/jvi.00848-26","url":null,"abstract":"<p><p>The diversity within H1 and H3 subtype influenza A viruses (IAVs) in swine prevents effective vaccine control approaches with inactivated whole-virus vaccines. We addressed the challenge of controlling co-circulating hemagglutinin (HA) clades of swine IAV with the development of a multivalent messenger RNA (mRNA)-lipid nanoparticle (LNP) vaccine expressing eight HA proteins to maximize genetic coverage. We applied a computational approach to select eight HA genes that represented 95% of the observed IAVs detected in the United States between 2022 and 2025. Piglets were vaccinated and boosted intramuscularly with either individual HA mRNA-LNP or an 8-HA multivalent mRNA-LNP. Serum was collected to evaluate systemic antibody levels. Twenty-one days post-boost, pigs were challenged with a field-relevant H1 1A.3.3.3-c3 IAV strain. The 8-HA multivalent mRNA-LNP vaccine induced neutralizing antibodies against all eight antigens, and vaccinees were protected against lung lesions, with lesion scores similar to non-challenged animals. Homologous monovalent vaccination significantly reduced IAV detection in nasal secretions and in the lungs. Heterologous monovalent vaccination was not cross-protective but did not induce vaccine-associated enhanced respiratory disease. We provide evidence that monovalent and multivalent mRNA-LNP influenza vaccines elicited neutralizing antibody responses in pigs and protected against viral challenge. The versatility and capacity for rapidly updating the mRNA-LNP vaccine platform make it an appealing tool to improve animal health and minimize the circulation and diversity of IAV in swine.IMPORTANCEInfluenza A virus (IAV) is an important respiratory pathogen in swine, and zoonotic transmission of swine strains to humans remains a public health risk. Control strategies against IAV in swine herds rely heavily on biosecurity measures and vaccination. However, the antigenic diversity of IAV circulating in swine challenges current vaccination programs, and there is a need for broadly protective vaccines or platforms that can rapidly update components to reflect circulating diversity. mRNA-LNP vaccines have emerged as promising vaccine platforms, offering simultaneous delivery of multiple antigens, rapid development, scalable manufacturing, and potent immunogenicity. In this study, we assessed the immunogenicity and protective capacity of monovalent and multivalent mRNA-LNP vaccines encoding eight representative IAV hemagglutinin (HA) antigens. To our knowledge, this is the first study to objectively select multiple representative endemic swine IAV strains by quantifying genetic diversity within the phylogeny and to apply this selection to rationally design and evaluate a multivalent HA mRNA-based influenza vaccine in the swine model.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0084826"},"PeriodicalIF":4.1,"publicationDate":"2026-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149137172","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":"H5 influenza virus mRNA-lipid nanoparticle (LNP) vaccination elicits adaptive immune responses in Holstein calves.","authors":"Carine K Souza, Jefferson J S Santos, Paola Boggiatto, Haley Sterle, Bailey Arruda, Mitchell V Palmer, Alessandra Campos, Jiaojiao Liu, Naiqing Ye, Drew Weissmann, Scott E Hensley, Amy L Baker","doi":"10.1128/jvi.01289-26","DOIUrl":"https://doi.org/10.1128/jvi.01289-26","url":null,"abstract":"<p><p>Highly pathogenic avian influenza (HPAI) clade 2.3.4.4b H5N1 circulated widely in lactating dairy cows in the United States following the initial detection in 2024. Due to the critical need for intervention strategies for this outbreak, we evaluated antibody and cellular immune responses of a clade 2.3.4.4b H5 mRNA-LNP vaccine in calves. We found that the H5 mRNA-LNP vaccine induced a robust antibody and CD8⁺ T cell-mediated immune response and conferred protection against clade 2.3.4.4b H5N1 infection.IMPORTANCEIt is important to develop vaccines that limit clade 2.3.4.4b H5N1 infection in cattle. In this study, we found that a new H5 mRNA-lipid nanoparticle vaccine is immunogenic when delivered intramuscularly to male Holstein calves.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0128926"},"PeriodicalIF":4.1,"publicationDate":"2026-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149137199","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}