PLoS Pathogens最新文献

{"title":"Viral interferon antagonism shapes host tropism.","authors":"Felix Streicher, Amandine Chantharath, Nolwenn Jouvenet","doi":"10.1371/journal.ppat.1013544","DOIUrl":"https://doi.org/10.1371/journal.ppat.1013544","url":null,"abstract":"<p><p>Outbreaks of zoonotic viruses in human populations highlight the need to understand the molecular factors that influence viral host tropism and interspecies transmission. A virus's host range is determined not only by the expression of host factors that facilitate viral entry and replication, but also by the virus's ability to evade host antiviral immunity. The interferon (IFN) response is a potent immune defense rapidly mobilized upon viral infections. To overcome this immune barrier and establish infection, viruses continuously evolve evasion strategies. This review illustrates, through a series of examples, how species-specific interactions between viruses and the IFN response erect interspecies barriers. We discuss the necessity to develop cellular models derived from viral reservoir hosts, as well as machine learning approaches, to better grasp species-specific viral sensitivity to the IFN response. These approaches will be essential for understanding viral interspecies transmission and guiding effective pandemic preparedness measures.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013544"},"PeriodicalIF":4.9,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145245732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus.","authors":"Jennifer McIntyre, Alison Morrison, Kirsty Maitland, Eileen Devaney, James A Cotton, Collette Britton, Ray Kaplan, Dave Bartley, Roz Laing","doi":"10.1371/journal.ppat.1013578","DOIUrl":"https://doi.org/10.1371/journal.ppat.1013578","url":null,"abstract":"<p><p>Subtherapeutic treatment or 'underdosing' is considered a common problem in the control of parasitic helminths of animals and people and can hasten the emergence of anthelmintic resistance. Increasing reliance on the long-acting macrocyclic lactone, moxidectin, in both veterinary and medical settings may increase exposure of incoming helminth populations to subtherapeutic drug concentrations due to its extended half-life. However, we lack genetic markers to monitor emerging resistance as the mechanism(s) underlying resistance to the macrocyclic lactones are unresolved in parasitic helminths. Furthermore, the impact of prior ivermectin exposure on the evolution of moxidectin resistance is unclear. To test the impact of subtherapeutic selection on the emergence of macrocyclic lactone resistance, we exposed a fully drug susceptible isolate of an economically important parasitic helminth of livestock, Haemonchus contortus, to low but increasing doses of ivermectin or moxidectin in vivo for phenotypic, genomic, and transcriptomic analyses. After a single subtherapeutic dose of ivermectin or moxidectin, we find evidence of selection at a shared genetic locus on Chromosome V, with the signal of selection increasing with subsequent doses. After only three subtherapeutic treatments, ivermectin-selected lines were resistant to a full standard (label) dose of ivermectin. However, moxidectin selected lines remained susceptible to a half dose of moxidectin. This was despite showing higher resistance to ivermectin in vitro and a stronger signal of selection at the Chromosome V locus than the equivalent ivermectin-selected lines. Our findings highlight the rapid selection for anthelmintic resistance with subtherapeutic treatment and implicate the pre-existence of ivermectin and moxidectin resistance haplotypes in a drug-naïve population. We demonstrate that ivermectin selected lines show emerging moxidectin resistance, underpinned by a shared genetic locus of resistance. Finally, we speculate that key differences in the resistance phenotype between ivermectin and moxidectin selected lines relate to differences in the inheritance of resistance within this shared locus, with ivermectin resistance manifesting as dominant trait while moxidectin resistance appears to be recessive.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013578"},"PeriodicalIF":4.9,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dysplastic lung repair fosters a tuberculosis-promoting microenvironment through maladaptive macrophage polarization.","authors":"Shivraj M Yabaji, Suruchi Lata, Anna E Tseng, Prasanna Babu Araveti, Ming Lo, Igor Gavrish, Aoife K O'Connell, Hans P Gertje, Anna C Belkina, Colleen E Thurman, Hirofumi Kiyokawa, Darrell Kotton, Shumin Tan, Janice J Endsley, William R Bishai, Nicholas Crossland, Lester Kobzik, Igor Kramnik","doi":"10.1371/journal.ppat.1013563","DOIUrl":"https://doi.org/10.1371/journal.ppat.1013563","url":null,"abstract":"<p><p>Pulmonary TB that develops in immunocompetent adult humans is responsible for approximately 85% of the disease burden and is central for Mtb transmission. Most humans contain Mtb infection within primary granulomatous lesions, but in certain immunocompetent humans, containment fails, leading to hematogenous spread and active pulmonary disease with the formation of cavities that enable Mtb transmission via aerosols. To reveal lung-specific microenvironments conducive for Mtb survival and replication despite systemic immunity, we use fluorescence multiplex immunohistochemistry and spatial transcriptomic analyses of heterogenous TB lesions that uniquely form in the lungs of immunocompetent but TB-susceptible B6.Sst1S mice after hematogenous spread from the primary lesion. Initially, these secondary lung lesions manifested local adoptive immunity featuring tertiary lymphoid follicles similar to resistant B6 mice and contained primarily non- replicating bacilli. Following these early events, however, the B6.Sst1S mice uniquely demonstrate expansion of myeloid cell populations with the appearance of alternatively activated macrophages, dissolution of lymphoid follicles, and the accumulation of de- differentiated lung epithelial cells. These processes led to bronchogenic expansion, broncho- occlusion, and necrosuppurative pneumonia closely resembling advanced pulmonary tuberculosis in humans. To determine whether lung parenchymal cells or lung oxygenation were necessary for the pulmonary TB progression, we implanted lung and spleen fragments subcutaneously prior to the infection. The lung implants uniquely displayed the formation of the characteristic organized granulomas with necrosis and Mtb replication that paralleled TB progression in native lungs, demonstrating that the cellular composition of inflamed lung tissue, not oxygenation, is a critical determinant of pulmonary TB progression. Our data demonstrate that deleterious bi-directional interactions of aberrantly activated macrophages with the inflammation-injured lung resident cells determine lung vulnerability to virulent Mtb in immunocompetent hosts. Because these mechanisms enable Mtb transmission among humans via aerosols, they are likely evolutionary conserved and, therefore, represent appealing targets for host-directed TB therapies.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013563"},"PeriodicalIF":4.9,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Re-emergence of chikungunya virus in China by 2025: What we know and what to do?","authors":"Tong-Yun Wang, Yue Sun, Yan-Dong Tang","doi":"10.1371/journal.ppat.1013556","DOIUrl":"10.1371/journal.ppat.1013556","url":null,"abstract":"<p><p>In July 2025, China witnessed its most significant chikungunya virus (CHIKV) outbreak since 2010. As of August 1, with over 6,000 cases reported in Foshan city, Guangdong Province. Although the clinical manifestations have been relatively mild, the rapid transmission within communities warrants our attention. In this context, we emphasize our current knowledge and the necessary actions to take. Specifically, we identify critical gaps in CHIKV control efforts and assess the effectiveness of current measures. These include vector management strategies, viral genomic surveillance, the deployment of newly approved vaccines, and the development of antiviral agents. Overall, to effectively control the epidemic of CHIKV, we require a comprehensive and multifaceted strategy for its prevention and management.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013556"},"PeriodicalIF":4.9,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12500152/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240061","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The serine protease HtrA regulates Group B Streptococcus virulence and affects the host response to infection.","authors":"Alyssa Brokaw, Grace Wallen, Austyn Orvis, Hei Joon Kwon, Ravin Seepersaud, Shayla Nguyen, Kavita Sharma, Michelle Coleman, Phoenicia Quach, Joy Twentyman, Jay Vornhagen, Lisa A Jones, Chenwei Lin, Philip R Gafken, Lakshmi Rajagopal","doi":"10.1371/journal.ppat.1013562","DOIUrl":"https://doi.org/10.1371/journal.ppat.1013562","url":null,"abstract":"<p><p>Group B Streptococcus (GBS) rectovaginally colonizes up to 20% of women worldwide and is a leading cause of invasive infections during pregnancy, contributing annually to a significant proportion of preterm births, neonatal infections, and stillbirths. Despite its reputation as a perinatal pathogen, GBS infection rates in non-pregnant adults are also increasing. While much progress has been made to understand transcriptional regulation of virulence by two-component systems, many aspects of GBS virulence regulation remain understudied. Although many bacterial pathogens utilize high temperature response A (HtrA) family serine proteases to regulate virulence and stress responses through varied mechanisms, the function of HtrA in GBS was unknown. Here, we demonstrate that HtrA is localized to the GBS membrane and regulates the abundance of endogenous surface and secreted proteins, including a subset of virulence factors. Although deletion of htrA (ΔhtrA) increased dissemination to placentas and fetuses, this strain caused significantly fewer adverse pregnancy outcomes compared to isogenic wild-type (WT). Placentas from ΔhtrA-infected dams contained more chemokines, pro-inflammatory IL-1β, and neutrophil myeloperoxidase than isogenic WT-infected placentas, suggesting that ΔhtrA GBS induces potent neutrophil chemotaxis. However, immunosuppressive IL-10 was present at increased concentration, which may in part explain the attenuation of adverse pregnancy outcomes during ΔhtrA infection. Finally, we note that recombinant GBS HtrA directly cleaves human fibronectin in vitro, highlighting that this protease may also target host substrates during infection. Together, these findings support a role for HtrA as a post-translational regulator of GBS virulence and suggest that inhibiting HtrA activity may hold therapeutic promise against GBS induced adverse pregnancy outcomes.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013562"},"PeriodicalIF":4.9,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Carabrone inhibits Gaeumannomyces tritici growth by targeting mitochondrial complex I and destabilizing NAD⁺/NADH homeostasis.","authors":"Xingyu Ren, Jing Bai, Yingying Han, Jiaying Xu, Yingchen Liu, Zhiqing Ma, Yong Wang, Juntao Feng","doi":"10.1371/journal.ppat.1013567","DOIUrl":"https://doi.org/10.1371/journal.ppat.1013567","url":null,"abstract":"<p><p>The excessive and irrational use of commercial fungicides has led to escalating drug resistance in phytopathogens, necessitating the discovery of novel antifungal targets and strategies. Plant secondary metabolites, serving as natural chemical defenses against pathogen invasion, offer promising scaffolds and potential targets for developing innovative crop protection approaches. This study elucidates the antifungal mechanism of the natural sesquiterpene lactone carabrone against Gaeumannomyces tritici through integrated multi-omics analyses. Time-series transcriptomic profiling revealed that carabrone significantly suppresses the oxidative phosphorylation (OXPHOS) pathway and disrupts nicotinate/nicotinamide metabolism, resulting in a reduced NAD⁺/NADH (NAD+, Oxidized nicotinamide adenine dinucleotide; NADH, Reduced nicotinamide adenine dinucleotide) ratio. Orthogonal elevation of NAD⁺ levels through exogenous supplementation diminished fungal susceptibility to carabrone, establishing a direct link between NAD⁺/NADH homeostasis and its antifungal activity. Activity-based protein profiling (ABPP), gene silencing screens, and physiological-biochemical validations collectively demonstrated that carabrone specifically inhibits the electron transport chain (ETC) rather than ATP synthase to regulate NAD⁺/NADH balance. Further evidence from pyruvate supplementation, expression of the yeast non-proton-pumping NADH dehydrogenase ScNDI1, and enzymatic assays confirmed that carabrone directly targets mitochondrial respiratory chain complex I, thereby destabilizing NAD⁺/NADH homeostasis and suppressing G. tritici growth. This work first establishes complex I as the direct antifungal target of carabrone, revealing its lethal mechanism involving complex I inhibition-mediated blockade of NADH oxidation, followed by oxidative stress induction and energy metabolism collapse. Additionally, we demonstrate that ScNDI1 serves as a critical tool for screening and validating complex I-targeted fungicides. These findings provide both a lead scaffold for developing novel complex I inhibitors and a systematic framework for antifungal agent validation, offering theoretical support to combat emerging fungal resistance challenges.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013567"},"PeriodicalIF":4.9,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145226087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antibiotic-induced morphological changes enhance phage predation.","authors":"Julián Bulssico, Swapnesh Panigrahi, Mélanie Matveeva, Nicolas Ginet, Mireille Ansaldi","doi":"10.1371/journal.ppat.1013546","DOIUrl":"https://doi.org/10.1371/journal.ppat.1013546","url":null,"abstract":"<p><p>Due to the high public health risk posed by antibioresistance, phage therapy - the use of bacteriophages as antibacterial agents - is experiencing renewed interest. As the combined administration of antibiotics and phages is a common practice in compassionate treatments, our research focuses on the effects of antibiotics on phage predation, which may be of crucial importance for phage therapeutic applications. A distinctive manifestation of phage infection in solid media is the appearance of lysis plaques, corresponding to the circular thinning of a bacterial lawn. During plaque formation, successive cycles of phage replication take place from a single point of infection and spread radially in a lawn of immobilized bacterial hosts. Many factors affect plaque size, such as the composition and the reticulation of the propagation matrix, the characteristics of the phage, but also parameters related to the physiology of the bacterial host. It has also been known for decades that some antibiotics enable phages to spread more rapidly, resulting in better bacterial eradication. This phenomenon, called Phage-Antibiotic Synergy (PAS), is evidenced by larger lysis plaques on solid media. Our previous experimental work has focused on the phage characteristics and pointed to enhanced adsorption as a key factor leading to more efficient predation. However, since sublethal antibiotic concentrations can drastically affect bacterial physiology - for instance halting cell division in the case of ciprofloxacin or ceftazidime - and since plaque formation is strongly influenced by host growth dynamics, a comprehensive model integrating both the host growth and phage infection parameters is required to investigate PAS. We characterized the epidemics of two different phages (T5 and T7) during E. coli MG1655 infection on semi-solid media in the presence of sublethal antibiotic concentrations that affect (or not) cell morphology in different ways (cell filamentation or cell bloating). We observed that in these conditions lysis plaque enlargement is linked to the host's morphological changes. We conclude this work with a mathematical model that captures such observations and explains the increase in plaque size observed in the presence of antibiotics.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013546"},"PeriodicalIF":4.9,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145226158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-cell analysis reveals an important role of CD137L+ macrophages in the host response to uropathogenic Escherichia coli infection in the bladder.","authors":"Yaxiao Liu, Zizhuo Yang, Yinrui Xiang, Guangzhou Cheng, Lipeng Chen, Shuai Wang, Maolin Zang, Nan Zhou, Xiaoyi Zhang, Rui Chen, Benkang Shi, Yan Li","doi":"10.1371/journal.ppat.1013543","DOIUrl":"10.1371/journal.ppat.1013543","url":null,"abstract":"<p><p>Uropathogenic Escherichia coli (UPEC) typically trigger rapid and robust innate immune responses in the bladder. In order to identify the key facets of the host response that influence pathogen clearance and tissue damage, single-cell RNA sequencing was used to investigate the transcriptomic changes of immune cells in mouse bladder after UPEC infection. Single-cell analysis revealed significant elevated CD137L expression in macrophages and dendritic cells in bladder after UPEC infection. CD137L defines a macrophage population in bladder that is important for the host response to UPEC infection. Deletion of CD137L in macrophages resulted in severe bacterial burden and bladder inflammation during the acute stage of UPEC infection. Further study demonstrated that the crucial role of CD137L+ macrophages in protecting against UPEC infection might be mediated by Tregs, which express high levels of CD137 (the receptor for CD137L). Deletion of CD137L+ macrophages decreased Treg cells and led to a reduction in inhibitory factors such as CTLA-4 and PD-1 on Tregs. Deletion of Tregs using Foxp3DTR mice also aggravated inflammatory reactions, bacterial load, and urothelial destruction during the acute phase of UPEC infection. Similarly, the deletion of CD137 in Tregs resulted in a decrease in these inhibitory factors on Tregs, causing more severe bladder inflammation during UPEC infection. These results illuminate the immune landscape of the bladder infected by UPEC and highlight the crucial role of CD137L+ macrophages during UPEC infection in bladder. CD137L+ macrophages might prevent excessive inflammatory response during the host response to UPEC infection by regulating Tregs.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013543"},"PeriodicalIF":4.9,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12494246/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145226074","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Host- and microbial-mediated mucin degradation differentially shape Pseudomonas aeruginosa physiology and gene expression.","authors":"Sabrina J Arif, Kayla M Hoffman, Jeffrey M Flynn, Talia D Wiggen, Sarah K Lucas, Alex R Villarreal, Adam J Gilbertsen, Jordan M Dunitz, Ryan C Hunter","doi":"10.1371/journal.ppat.1013568","DOIUrl":"10.1371/journal.ppat.1013568","url":null,"abstract":"<p><p>Pseudomonas aeruginosa is a hallmark pathogen of cystic fibrosis (CF) airway infections, capable of reaching high cell densities despite its limited ability to directly utilize mucin glycoproteins as a nutrient source. In the CF lung, however, P. aeruginosa may access preferred carbon sources (e.g., amino acids and short-chain fatty acids) through metabolic cross-feeding with co-colonizing mucin-degrading microbes. Although host-derived enzymes such as neutrophil elastase can also degrade mucins, the extent to which host-mediated mucin breakdown supports P. aeruginosa growth remains unclear. Thus, here we compared the nutritional impact of microbial versus host mucolytic activity on P. aeruginosa physiology. Analyses of CF sputum revealed patient-specific variability in mucin integrity that is shaped by both host and microbial factors. We demonstrate that mucin degradation by anaerobic bacteria through proteolysis, glycolysis, and fermentation, promotes robust P. aeruginosa growth, unlike mucin processed by neutrophil elastase alone. Targeted metabolomics identified acetate and propionate as key metabolites driving this cross-feeding, while transcriptomic and phenotypic analyses revealed that P. aeruginosa engages in diauxic growth on a broader set of mucin-derived substrates. Unexpectedly, cross-feeding with anaerobes triggered the induction of P. aeruginosa denitrification and fermentation pathways, suggesting redox remodeling despite being cultured under oxygen-replete conditions. Finally, the transcriptional profile of P. aeruginosa grown on anaerobe-conditioned mucins more closely resembled its in vivo gene expression, more so than when grown on intact or neutrophil-degraded mucins. Together, these findings provide new insight into the potential role of interspecies metabolic interactions in shaping pathogen physiology in the inflammatory, polymicrobial, and mucus-rich environment of the CF airways.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013568"},"PeriodicalIF":4.9,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145226080","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"West Nile virus vaccine candidates attenuated by dinucleotide enrichment are immunogenic and protective against lethal infection.","authors":"Nguyen Phuong Khanh Le, Prince Pal Singh, Ivan Trus, Uladzimir Karniychuk","doi":"10.1371/journal.ppat.1013560","DOIUrl":"https://doi.org/10.1371/journal.ppat.1013560","url":null,"abstract":"<p><p>West Nile virus (WNV) poses a global public health threat. This study demonstrates that the WNV RNA tolerates CpG and UpA dinucleotide enrichment in different genomic regions resulting in attenuation of CpG- and CpG/UpA-enriched variants. Attenuation was zinc finger antiviral protein 1 (ZAP)-dependent, and ZAP knockout (ZAP-KO) cells were used to generate high-titer stocks. Ten enriched variants, with permuted control and wild-type (WT) viruses, were screened in immunocompetent mice upon intraperitoneal injection. In contrast to lethal WNV-WT and permuted viruses, the E-MAX variant, with the RNA region encoding envelope (E) protein enriched both with CpG and UpA, caused no mortality. E-MAX was immunogenic and protective against lethal challenge. Stability of enriched dinucleotides was confirmed upon serial passaging in ZAP-WT and ZAP-KO cells, with only minor (17-21%) reversion at a single site in ZAP-WT condition. E-MAX upregulated interferon (IFN) signaling genes in human cells, suggesting that the combination of CpG/UpA-mediated attenuation, and concurrent activation of IFN responses potentially driven by CpG/UpA enrichment, may contribute to E-MAX immunogenicity. Evaluation using footpad injection in mice showed E-MAX had a promising safety and immunogenicity profile, although brain infection was occasionally detected. Then, we developed the E-MAX+ FR variant by combining CpG/UpA enrichment with two amino acid substitutions in functional domains of the E protein. This strategy eliminated neuroinvasion while maintaining immunogenicity and protection. Altogether, CpG/UpA dinucleotide enrichment in the genomic E region in combination with amino acid substitutions in the E protein yields a promising platform for vaccine development against WNV and potentially other flaviviruses.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013560"},"PeriodicalIF":4.9,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145226145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}