Background Human β-coronaviruses are a group of viruses that infect humans and cause upper respiratory tract diseases. Antigen detection could be used as an auxiliary diagnostic method for viral infection. The study aimed to develop a broadly cross-reactive antibody capable of simultaneously recognizing the nucleocapsid (N) proteins of five human β-coronaviruses, for use as a capture antibody for viral antigen detection. Methods The homologous sequence peptides of N proteins from five human β-coronaviruses were synthesized, coupled to carrier proteins, and used as immunogens to immunize New Zealand white rabbits. The antisera were purified using ammonium sulfate precipitation and antigen affinity chromatography. The capture performance of the NPA antibody for the N proteins of the five β-HCoVs was verified by sandwich ELISA and competitive ELISA. Throat swab specimens from SARS-CoV-2-infected patients were used to verify the capture performance of the NPA antibody against the N proteins in real-world clinical samples. Results The synthetic peptides were >95% pure as determined by high-performance liquid chromatography and their molecular weights were confirmed by mass spectrometry to match the theoretical molecular weights. The antibody titer of rabbit antiserums was determined to be 1:8000 and the purity of purified polyclonal antibody (NPA) was higher than 90%. The NPA antibody was capable of capturing the N proteins of the five β-HCoVs. When the NPA antibody was tested for N proteins capture in SARS-CoV-2 patient specimens, six of eight nucleic acid-positive specimens with cycle threshold (Ct) values < 30 tested positive. Conclusion The broadly cross-reactive antibody obtained in the study could be used as capture antibody in human β-coronavirus antigen detection and provides a solid technical foundation for other newly emerging β-coronavirus subspecies in the future.
Aphids are one of the most important groups of agricultural pests due to their ability to transmit a wide range of plant viruses. The development of more sustainable and specific methods for plant virus control is an ongoing and active avenue of research. In this study, we demonstrate by feeding dsRNA to aphids via artificial diet assay, the knockdown of two M. persicae (green peach aphid) stylet cuticle proteins (ST-1 and ST-2) implicated in the attachment and retention of the non-persistent, Potyviridae family member, zucchini yellow mosaic virus (ZYMV) (Potyvirus cucurbitaflavitesselati). We further enhanced ST-1 and ST-2 transcript knockdown by co-administering Nuclease dsRNA (NUC1/ -2_dsRNA) as an RNAi of RNAi approach, enhancing the survivability of orally co-delivered ST1 and ST2_dsRNA molecules against endonucleases in the aphid's midgut. Functionally, we successfully show significant knockdown of the ST1 and ST2 gene upon co-administered with Nuc1_dsRNA translates to a significant reduction of virus particle retention of up to 84% and 90%, respectively in aphids. Particle reduction translated to a 60 - 73% reduction in the infection rate when zucchini plants were challenged with treated aphids. This study demonstrates the successful application of an RNAi of RNAi approach to significantly disrupt the virus-vector interaction effectively reducing virus transmission to plants and laying a solid foundation towards effective multi-stage life cycle targeting with RNAi-based viricidal applications.
Rabies virus (RABV) is a zoonotic pathogen with an almost 100% case-fatality rate and limited post-exposure prophylaxis options. mRNA-based antigen delivery represents a promising approach but requires further optimization. We prepared a cationic lipid nanoparticle (LNP) delivery system using the thin-film hydration method and characterized its physicochemical properties and delivery efficiency. An mRNA encoding a fusion antigen comprising rabies virus glycoprotein (G) and herpes simplex virus type 2 infected cell polypeptide 35 (ICP35), lacking canonical 5' and 3' untranslated region sequences, was constructed. In vitro protein expression of the G-ICP35 mRNA was confirmed. Immunogenicity was evaluated in BALB/c mice following three intramuscular immunizations with G-ICP35 mRNA, RABV G/N proteins, or their combination. Tail vein blood samples were collected on days 7, 21, and 39 after the final immunization, and rabies virus-specific antibody titers and T-cell responses were assessed. The cationic LNP formulation efficiently mediated intracellular delivery and expression of the G-ICP35 mRNA. In vivo, the mRNA-LNP formulation induced detectable humoral and cellular immune responses, including rabies virus-specific IgG production and antigen-specific T-cell activation. Notably, co-immunization with mRNA and subunit vaccines further enhanced immune responses compared with either vaccine alone, achieving peak rabies virus-specific IgG titers of 694.5 IU/mL. These findings demonstrate the feasibility of a DCChol-based mRNA-LNP formulation for fusion antigen delivery and support further investigation of combined mRNA and subunit vaccine strategies.
Newcastle disease virus (NDV) remains one of the most economically important avian pathogens worldwide, causing recurrent outbreaks in poultry despite decades of vaccination and disease control efforts. Since the first reported outbreak of NDV a hundred years ago, numerous molecular epidemiological studies have been conducted globally across diverse geographic and production settings. Following a century of NDV circulation and evolution, the present study aimed to compile all publicly available NDV sequence data and perform a comprehensive global analysis of the genetic diversity, phylogenetic relationship, and global spatiotemporal distribution of NDV over a 100-year timescale. All publicly available NDV complete genome and full-length fusion (F) gene sequences were retrieved from GenBank up to February 2026. Following rigorous quality control, phylogenetic analyses were performed based on complete genomes and F gene datasets. Phylogenetic analysis identified two genotypes within Class I and 20 genotypes within Class II NDVs, with extensive diversification at the sub-genotype level. Genotype XIII exhibited the greatest sub-genotypic diversity, while genotype VII represented the most globally disseminated genotype, reported across 36 countries. Chronological assessment based on the earliest available reports indicated an increasing number of recognized genotypes from the 1930s to recently described sub-genotypes such as XIII.2.3 and XXII.2.2. Regional diversity analysis revealed the highest genotype diversity in Western Africa, Eastern Asia, and Southern Asia. Comparative residue analysis demonstrated substantial genotype-specific variation within critical functional domains of the fusion protein, including cleavage sites, neutralizing epitopes, and heptad repeat regions. Overall, this study provides the first comprehensive 100-year global overview of NDV evolution and phylogeography. The findings highlight continuous viral diversification, broad geographic dissemination of multiple genotypes, and ongoing molecular variation, emphasizing the need for sustained genomic surveillance and periodic evaluation of vaccine compatibility with emerging NDV genotypes.
Antimicrobial resistance (AMR) has escalated into a global health crisis, with resistant pathogens causing over 1.2 million direct deaths annually and threatening to render modern medicine unsustainable. This review provides a comprehensive and updated synthesis of CRISPR-Cas-based antimicrobial strategies with a unique focus on: (i) critical comparison with conventional antibiotics and emerging alternatives; (ii) quantitative evaluation of delivery platforms; (iii) novel strategies including AI-optimized guide design and the ATTACK-CreTA system; (iv) comprehensive analysis of ecological risks; and (v) technology readiness level assessments for clinical translation. The CRISPR-Cas system, originally discovered as a bacterial adaptive immune mechanism, has been repurposed as a programmable precision tool to combat AMR by selectively targeting and eliminating resistance genes. We systematically evaluate the mechanistic diversity of Cas effectors, from DNA-cleaving Cas9 and Cas3 to RNA-targeting Cas13, and their application in reversing resistance phenotypes in WHO priority pathogens. We critically assess emerging delivery platforms, including engineered bacteriophages, conjugative plasmids, nanoparticles, and outer membrane vesicles, quantitatively comparing their delivery efficiency, payload capacity, and biosafety profiles. Novel strategies such as CRISPR interference (CRISPRi) for gene silencing without genomic cleavage, the ATTACK-CreTA system for enhanced bactericidal activity, and AI-driven optimization of guide RNA design are examined with appropriate caveats. We comprehensively address clinical translation challenges including immunogenicity, pharmacokinetics/pharmacodynamics, manufacturing scalability, regulatory pathways, and bacterial resistance mechanisms including anti-CRISPR proteins. No CRISPR-based antimicrobial has yet received regulatory approval, and we critically evaluate the gap between proof-of-concept and clinical utility. A detailed roadmap for clinical development is proposed. By integrating recent advances in Cas protein engineering, delivery technologies, and diagnostic applications, this review positions CRISPR-Cas systems as next-generation precision therapeutics capable of both treating resistant infections and curtailing the spread of AMR across clinical and environmental settings.
The hepatitis B virus (HBV) X protein (HBx) functions as a versatile regulatory molecule that is indispensable for both efficient HBV replication and the pathogenesis of HBV-related hepatocellular carcinoma (HCC). Accumulating studies indicate that a broad spectrum of post-translational modifications (PTMs), including ubiquitination, phosphorylation, NEDDylation and ISGylation, critically influence HBx stability, intracellular trafficking, and its capacity to engage with diverse host factors. Collectively, these PTMs constitute a complex modulatory network that governs viral persistence and contributes to malignant transformation in the liver. In this review, we synthesize recent advances in deciphering the molecular principles that drive HBx PTMs and delineate their functional consequences during HBV infection and HCC development. We also summarize current understanding of how HBV manipulates host PTM systems to optimize its life cycle while promoting oncogenic processes. Furthermore, the review discusses emerging therapeutic opportunities centered on modulating HBx PTMs, with the aim of informing the design of next-generation antiviral and anti-tumor interventions.
Enterovirus A71 (EV-A71) causes hand, foot, and mouth disease and can trigger life-threatening neurological complications, yet the sequence-level physicochemical correlates of CNS involvement across globally circulating lineages remain incompletely defined. Here we screened 15,247 EV-A71 genomic entries spanning 1998–2024, retaining 267 full-length sequences (≥7,000 bp) with confirmed clinical outcomes (7 central nervous system [CNS]-involved, 260 non-CNS). This extreme 7:260 class imbalance, reflecting the scarcity of publicly available full-length CNS-associated EV-A71 genomes, is the principal limitation and interpretive premise of the study. Each polyprotein position was encoded by three Z-scale descriptors—hydrophobicity (Z1), molecular volume (Z2), and electrostatic polarity (Z3)—converting discrete residue identities into a continuous biophysical feature space. A two-stage statistical pipeline (Mann–Whitney U screening followed by odds-ratio ranking) distilled 20 significant loci down to five core positions: P2124_Z1, P997_Z2, P1246_Z3, P1743_Z2, and P1711_Z1 (all P<0.001). Leave-one-out cross-validated logistic regression achieved the highest area under the receiver operating characteristic curve (AUC = 0.889) among eight algorithms benchmarked. Because this AUC is estimated from only seven positive samples, it should be regarded as an exploratory internal performance signal rather than definitive evidence of generalisable accuracy. SHapley Additive exPlanations (SHAP) assigned the largest model contribution to P2124_Z1 (OR = 4.28; 95% CI 1.47–12.51), while P1246_Z3 was statistically associated with lower CNS odds (OR = 0.50); these model-derived quantities do not establish causal mechanisms. Reference-strain mapping linked the five polyprotein coordinates to mature-protein residues in 3D RdRp, 3C protease, 2C helicase, and 2A, thereby providing structural context for cautious biochemical hypotheses rather than confirmed mechanisms. Phylogenetic dispersion of CNS-associated strains was compatible with convergent evolution, but this inference remains limited by the seven available CNS genomes. We therefore present the five-position physicochemical signature and nomogram as hypothesis-generating tools for prioritising candidate neurovirulence markers, requiring prospective validation in larger and more balanced independent cohorts before clinical or field deployment.
Grapevine downy mildew, caused by the obligate biotrophic oomycete Plasmopara viticola, represents a major constraint to viticulture worldwide. Previous metatranscriptomic analyses revealed an unexpectedly rich viral diversity associated with downy mildew leaf lesions and in this study we re-analyzed the same RNA-seq libraries to complete viral genomes and identify additional RNA infectious elements. Using homology-based searches, RNA structural motif detection, coverage-based segment association and targeted experimental validation, we reconstructed the presumed complete genomes of multiple viruses belonging to the Splipalmiviridae, the proposed 'Orfanplasmoviridae' and 'Viviviridae' families. Specifically, we completed the genomes of seven known and three novel splipalmiviruses, five known and two novel orfanplasmoviruses, and four viviviruses, substantially expanding their genomic characterization. We further identified six novel ambiviruses, designated "downy mildew lesion-associated ambivirus 1-6" (DmlaAV1-6), which have a circular genome encoding two open reading frames on opposite polarities, including a putative RNA-dependent RNA polymerase. One DmlaAV contains paired delta-like ribozymes for which the self-cleavage activity was demonstrated in vitro, providing the first biochemical evidence of functional delta-like ribozymes in ambiviruses. In addition, a 767-nt circular RNA element lacking coding capacity and homology to known sequences was identified. This RNA adopts a quasi-rod-like structure, contains twister ribozymes in both polarity strands and lacks a DNA counterpart, supporting its classification as an infectious viroid-like RNA. Overall, this work refines and extends the virome associated with grapevine downy mildew leaf lesions, highlighting the remarkable diversity of RNA infectious agents inhabiting this ecological niche and underscoring the value of re-analyzing existing transcriptomic datasets.
Avian orthoreovirus (ARV) imposes substantial economic losses on the global poultry industry, primarily through runting–stunting syndrome and viral arthritis. Despite its global prevalence, the molecular epidemiology and pathogenic landscape of ARV strains circulating in Upper Egypt remain largely unexplored. This study investigated the molecular characteristics, phylogenetic relationships, and pathogenic potential of ARV in broiler flocks across Assiut, Sohag, and New Valley governorates. Out of 140 clinical samples from chickens showing stunted growth and tendon swelling, 42 (30%) were positive by RT-qPCR targeting the σC gene. Phylogenetic analysis of 11 representative isolates using MEGA X revealed a complex co-circulation of two genetically distinct genotypes. Notably, this study provides the first report of ARV Genotypes IV (GC IV) and V (GC V) in Upper Egypt governorates. Nine isolates clustered within Genotype V, forming a unique regional sublineage, while two isolates belonged to Genotype IV, showing high homology to contemporary Asian strains. Crucially, these field isolates exhibited extreme genetic divergence from commercial vaccine strains, sharing as little as 38% amino acid identity, indicating a severe vaccine mismatch. Experimental infection of young chicks confirmed high virulence, characterized by significantly impaired growth, reduced tibial length, and elevated feed conversion ratios (p <0.05). High viral loads were quantified in the tendons, pancreas, and proventriculus (p < 0.01). These findings underscore the urgent emergence of novel ARV variants in Upper Egypt and highlight the critical need for developing regionally-tailored vaccination strategies to mitigate these evolving threats.
Avian species play a key role in the ecology of viruses, acting as reservoirs, sentinels, and biological carriers across diverse environments. Here, we describe the detection and genomic characterization of a novel iflavirus identified in fecal samples from the Chiroxiphiapareola("blue-backed manakin") collected in areas of the Middle North region of Brazil, Maranhão state. Viral RNA was extracted from pool fecal samples and subjected to next-generation sequencing. De novo assembly and comparative analyses enabled the recovery of a complete picorna-like viral genome of 9043 nucleotides, comprising a single open reading frame encoding a polyprotein of 2896 amino acids. Phylogenetic analyses based on the RNA-dependent RNA polymerase (RdRp) domain and the translated polyprotein consistently clustered the virus within the family Iflaviridae, forming a specific clade with reference sequences previously reported in arthropods. Functional domain analysis revealed conserved motifs characteristic of positive-sense single-stranded RNA viruses, including helicase superfamily 3 and RdRp domains. Although iflaviruses are classically associated with arthropod hosts, their detection in avian fecal samples is likely related to dietary intake, suggesting the presence of a transient virome rather than active infection. The sampling area is characterized by increasing environmental degradation, which may favor interactions between wildlife and anthropogenic environments, highlighting the importance of viral surveillance. This study reports, for the first time, the genome of an iflavirus detected in C. pareola, expanding current knowledge on iflavirus diversity and reinforcing the relevance of wildlife-based surveillance in ecologically altered regions under anthropogenic pressure.
African swine fever (ASF) is an acute hemorrhagic disease caused by African swine fever virus (ASFV), with mortality rates approaching 100% in domestic pigs. Because its genome is large and structurally complex, the functions of many ASFV encoded genes remain poorly understood. In this study, ASFV I9R was identified as a relatively conserved early transcribed gene. To investigate its biological role, an I9R deletion recombinant virus (ASFV-ΔI9R) was generated by replacing the I9R gene with an enhanced green fluorescent protein (EGFP) expression cassette. In primary porcine alveolar macrophages (PAMs), ASFV-ΔI9R and the parental ASFV CN/GS/2018-WT (ASFV-WT) exhibited identical replication kinetics, indicating that I9R is dispensable for viral replication in vitro. Transcriptome sequencing (RNA-seq) analysis of infected PAMs at 18 and 36 hour post infection (hpi) showed that differentially expressed genes (DEGs) in ASFV-ΔI9R-infected cells were mainly enriched in innate immune signaling pathways, particularly pathways associated with type I interferon (IFN-β) mediated antiviral responses. Further analyses demonstrated that ASFV-ΔI9R infection reduced IFN-β-induced the expression of interferon-stimulated gene 12A (ISG12A) and inhibited phosphorylation levels of STAT1 and STAT2, key components of the JAK-STAT signaling pathway. Collectively, these findings suggest that I9R is involved in modulation of host antiviral responses through the JAK-STAT signaling pathway. This study provides additional insights into ASFV host interactions and contributes to understanding the biological functions of ASFV encoded genes.
Corneal infections with herpes simplex type 1 (HSV-1) are a leading cause of blindness in the world. At present the standard of care for corneal disease (herpetic stromal keratitis, HSK) is anti-virals and topical steroids. With the development of anti-viral resistant strains of the virus, it is important to discover new therapies for this disease. We have focused on the development of vaccines that would select for an immune response that controls both virus and corneal disease. As present most vaccines that have been studied tend to be efficacious when used in a prophylactic regimen. However, these vaccines are not effective when treating latently infected individuals when their disease is reactivated resulting in recurrent HSK. We decided to test the concept that if we were to include the co-stimulatory molecule CD137L in our vaccine construct that this would lead to success when used in a therapeutic paradigm. We present data indicating that this vaccine construct is effective in ameliorating both primary and recurrent forms of HSK. At this time, we cannot attribute this observation to specific mechanisms that may be mediating its protective effect, but that is something we wish to address in the future.
Beak and feather disease virus (BFDV, species Circovirus parrot) is the causative agent of psittacine beak and feather disease (PBFD), characterised primary by death of nestlings, feather loss and presumed immunosuppression in parrots worldwide. BFDV shows marked genetic heterogeneity, particularly in the open reading frame 2 (ORF2) encoding the capsid protein (Cap), but the impact of this diversity on antibody recognition has not been systematically assessed. In this study, we used seven recombinant Cap proteins derived from ORF2 variants selected to reflect the diversity of BFDV genotypes circulating in psittacine birds. N-terminally truncated, codon-optimised cap gene constructs were expressed in Escherichia coli and tested by semiquantitative Western blot and indirect ELISA using sera from eight naturally infected, BFDV-PCR-positive psittacine birds. Both assays revealed clear antigen-serum dependent differences in reactivity, with several sera failing to produce detectable signals against some Cap variants under the conditions used. These findings provide experimental evidence that amino acid variation in Cap can be associated with distinct antibody binding patterns in naturally infected birds. The observed heterogeneity in antigen recognition is likely to have direct implications for natural infections with BFDV, for the design and interpretation of BFDV serological assays and should be considered in future development of Cap-based vaccines.
Usutu virus (USUV) is an arbovirus that circulates between mosquitoes and birds with occasional spillover events into non-avian species. USUV continues to circulate in the Netherlands, particularly in Eurasian blackbirds (Turdus merula), although other USUV-susceptible avian species have also been found. Current serosurveillance systems make use of serological methods that can detect anti-USUV antibodies in sera collected from live birds. However, they require a dedicated laboratory and rarely include serological analysis of deceased avian specimens. As a proof of concept, this study evaluates the use of a previously developed lateral flow microarray immunoassay (LMIA) to detect USUV non-structural 1 (NS1)-specific antibodies in blood clots and lung and liver tissue extracts collected from blackbird carcasses found in the Netherlands. The likelihood of detection of USUV NS1 antibodies in USUV RT-PCR-positive blackbirds was found to trend higher in lung tissue extracts. Additionally, the LMIA exhibited only minimal cross-reactivity between USUV NS1-specific antibodies and USUV NS1 and West Nile virus (WNV) NS1 capture antigens on the LMIA. This initial study demonstrates the feasibility of the LMIA to specifically detect USUV NS1 antibodies in lung tissue extracts of deceased blackbirds. In future, a serological tool such as the one presented here, would allow for the rapid on-site analysis of both live and deceased blackbird specimens found in the field.
Enterovirus A71 (EV-A71) is a major causative agent of hand, foot, and mouth disease (HFMD) with neurotropism, and currently no approved antiviral therapies are available. In this study, we found that the ondansetron exhibits antiviral activity against EV-A71 infection. Ondansetron significantly reducing viral RNA levels, progeny virus production, and VP1 protein expression in a dose-dependent manner. In vivo, ondansetron treatment increased survival of neonatal mice and decreased viral loads in multiple tissues. Mechanistically, ondansetron modulates cholesterol metabolism-related genes to affect cell cholesterol stability and interferes with viral attachment during the early stage of infection, thereby inhibiting EV-A71 infection. These findings indicate that ondansetron is a promising candidate for drug repurposing to treat EV-A71 infection.
Numerous studies have examined the factors contributing to the variation in disease progression among people living with HIV-1, identifying a range of genetic, immunologic and virologic factors that interact in a complex, multifactorial dynamic. However, investigating individual susceptibility to HIV-1 infection adds an additional layer of complexity. In this observational report, we focus on the case of a man (Case-2021) who previously remained HIV-1 negative despite 35 years of self-reported high-risk sexual behavior with other men while enrolled in the MACS/WIHS Combined Cohort Study (MWCCS). To understand why this individual stayed free of HIV-1 infection for so long and what led to his eventual seroconversion, we examined the available genetic, immunologic, and virologic laboratory data before and two weeks after his diagnosis of HIV-1 infection and initiation of antiretroviral therapy (ART). In addition, we looked into two other high-risk men from same group (HR-10) who have sex with men (MSM) enrolled in the MWCCS who seroconverted later in their enrollment. Furthermore, we performed similar laboratory tests to assess the effect of seasonal variations on blood biomarkers from the same clinical visit as Case-2021, including one individual who already was living with HIV-1 and another who has remained seronegative throughout their enrollment in the MWCCS. In conclusion, this Case-2021 observational report suggests that resistance to HIV-1 infection is likely multifactorial, involving high risk sexual behavior, HIV-1 variants, the genomic structure of the CCR5 receptor on CD4+ T cells, and age-related immune changes. Further study is needed to investigate both the genomic protective factors of CCR5 against HIV-1 infection and these age-related immune changes.
Viruses are evolving biological entities whose perpetuation depends on their adaptive response to many selection pressures acting on them. Viruses are also solid objects that show some degree of deformability, brittleness, and strength when subjected to mechanical forces. Atomic force microscopy (AFM) allows the controlled application of mechanical force on individual virions, capsids, and viral components under close to physiological conditions. In this way, the structure, dynamics, and mechanical properties of virus particles, and their changes by the action of biologically relevant effectors, can be investigated at the single-particle level. AFM-based studies have shown that the mechanical properties of some viruses can have an adaptive value for virus survival, that mechanical forces are among the selection pressures some viruses may be confronted with, and that some viruses can exert mechanical forces that also constitute adaptive traits. AFM studies of viruses under mechanical force are also providing new insights into the relationships between virus structure and dynamics, physical properties, and biological function, even in those cases where natural mechanical forces may not be involved. The knowledge being acquired on virus biomechanics is contributing to understand virus biology and to the development of new biomedical or nanotechnological applications. The present review is addressed to a general readership of virologists, and attempts to summarize the results of over 20 years of studies that have used AFM, in combination with other biophysical, biochemical and/or biological approaches, to explore the connections between virus biology and mechanical force.
Epstein-Barr virus (EBV) is a ubiquitous herpesvirus associated with a broad spectrum of malignancies and immune-mediated disorders, and growing evidence highlights the importance of host glycan-lectin interactions in shaping viral persistence and immune escape. Among these, galectins have emerged as key regulators of the EBV life cycle, influencing viral attachment, latency maintenance, lytic reactivation, and the remodeling of the tumor microenvironment. Galectin-1, -3, and -9 exhibit context-dependent functions that collectively modulate oncogenic signaling pathways, T‑cell exhaustion, regulatory T‑cell expansion, and innate immune sensing. Recent clinical studies further suggest that circulating galectins and galectin-enriched exosomes may serve as non-invasive biomarkers for disease progression and prognosis in EBV-associated malignancies. Despite these advances, major knowledge gaps remain regarding member-specific functions, compensatory galectin networks, and the spatiotemporal dynamics of galectin regulation during infection. Targeting the galectin-glycan axis therefore represents a promising frontier for host-directed antiviral and anticancer therapies, with the potential to disrupt viral latency, restore antiviral immunity, and improve clinical outcomes in EBV-driven diseases.
The persistence of latent HIV-1 reservoirs poses a major risk for rebound viremia, necessitating strategies that enhance the clearance of intracellular viral components. Autophagy is a critical homeostatic mechanism for clearance of damaged organelle and misfolded proteins, yet its role in HIV-1 infection is complex, as the virus often evolves mechanisms to evade autophagic degradation. In this study, we identify the lysosomal transporter SLC29A3/ENT3 as a key regulator of viral clearance. Using SLC29A3-knockdown (KD) and overexpressing (OE) HEK293-T cell models, we demonstrate that the loss of SLC29A3 significantly increases susceptibility to infection by both HIV enveloped (Clade B and C) and VSV-G enveloped HIV-1 viral particles. SLC29A3-deficient cells exhibited higher rates of host genome integration and prolonged retention of the viral capsid protein p24. Immunofluorescence microscopy revealed that the absence of SLC29A3 leads to the persistent accumulation of viral cores within endolysosomes. Conversely, SLC29A3 replenishment expedited viral clearance; a process confirmed to be autophagy-dependent through pharmacological modulation with rapamycin and chloroquine. Our findings suggest that augmenting SLC29A3 activity may provide a novel therapeutic avenue for curbing viral load by enhancing virophagy.
