Harboring four transmembrane domains in their structural hallmark, Tetraspanins (Tspans) are a family of glycoproteins with pivotal functions in a variety of biological and cellular processes. Through interacting laterally with each other or specific membrane proteins, Tspans organize tetraspanin-enriched microdomains (TEMs), modulating cellular signaling, adhesion, fusion, and proliferation. An abundance of evidence has identified the multiple functions in the progression of cancer as well as the underlying molecular mechanisms. Recently, plenty of studies have focused on the utilities of Tspans by pathogens for infection, especially the infection of viruses. The expression of Tspans correlates with the phase of viral infection, the type of virus, and targeted therapies. In particular, perturbations of Tspans in host cells can affect viral attachment, intracellular trafficking, translation, virus assembly, and release. In this review, we summarize and provide a historical overview of the discovery and characterization of various kinds of virus infection and highlight their diversity and complexity, along with the virus life cycle. Furthermore, we examined the current understanding of how various Tspans are involved in the regulatory mechanisms underlying viral infection. This review aims to offer a comprehensive understanding of the targeting of Tspans for therapeutic intervention in infections caused by diverse pathogens.

is an important zoonotic pathogen that threatens human and pig health. During infection, the host can impose oxidative stress to resist pathogen invasion. Resistance to oxidative toxicity is an important factor for pathogens. Glutathione synthesis contributes to reactive oxygen species (ROS) detoxification in bacterial cells. Little is known about the roles of glutathione synthesis and transport in . . In this study, we demonstrated that glutathione treatment increased oxidative stress tolerance in . . GshAB and GshT were found in . glutathione synthesis and import by bioinformatics. In vitro, inactivation of and led to increased sensitivity to oxidative stress. Inactivation of led to growth defects in the medium. The intracellular glutathione content of or deletion mutants was lower than that of wild type (WT) strain. The phagocytic resistance of and mutants was lower than that of the WT strain. Moreover, the virulence of and deletion mutants was significantly lower than that of the WT strain in mouse survival and tissue loading experiments. In conclusion, these results revealed the functions of GshAB and GshT in the pathogenesis of . These findings enhance our understanding of bacterial virulence mechanisms and may provide a new avenue for therapeutic intervention aimed at curbing infections.

As antibiotic resistance escalates into a global health crisis, novel therapeutic approaches against infectious diseases are in urgent need. , an adaptable opportunistic pathogen, poses substantial challenges in treating a range of infections. The quorum-sensing (QS) system plays a pivotal role in orchestrating the production of a large set of virulence factors in a cell density-dependent manner, and the anti-virulence strategy targeting QS may show huge potential. Here, we present a comprehensive investigation into the potential of the synthesized compound 3-(benzo[d][1,3]dioxol-4-yl)oxazolidin-2-one (OZDO, CHNO) as a QS inhibitor to curb the virulence of . By employing an integrated approach encompassing screening, and functional identification, we elucidated the multifaceted effects of OZDO. Molecular docking predicted that OZDO interfered with three core regulatory proteins of QS system. Notably, OZDO exhibited significant inhibition on the production of pyocyanin, rhamnolipid and extracellular proteases, biofilm formation, and cell motilities of . Transcriptomic analysis and quantitative real-time PCR displayed the down-regulation of QS-controlled genes in OZDO-treated PAO1, reaffirming the QS-inhibition activity of OZDO. assessments using a -infection model demonstrated OZDO mitigated pathogenicity, particularly against the hypervirulent strain PA14. Moreover, OZDO in combination with polymyxin B and aztreonam presented a promising avenue for innovative anti-infective therapy. Our study sheds light on the multifaceted potential of OZDO as an anti-virulence agent and its significance in combating -associated infections.

Over the last decade, there has been a steady increase in HIV-1 prevalence in Mizoram, India. Importantly, this increase in HIV-1 prevalence is not only limited to the key population groups such as female sex workers (FSWs) or people who inject drugs (PWID), and has been witnessed in general population as well. Injecting drug use has long been one of the key drivers of HIV-1 epidemic across the north-eastern states of India. In this study, using HIV-1 pol gene region sequences from Aizawl and adjoining districts, we examined the HIV-1 subtypes, recombinant forms, drug resistance mutations and also the spatiotemporal dynamics of the potential unique recombinant forms. In our dataset, the dominant subtype was HIV-1 subtype C (94.91%). We could also identify the presence of CRF01_AE (1.69%) and BC recombinant forms (3.39%). Drug resistance mutation analysis revealed that resistance against non-nucleoside reverse transcriptase inhibitors was most common in the sequences having any resistance mutations. Evolutionary analysis of unique BC recombinants estimated the most recent common ancestor of these sequences around 2004-2005 and them having an ancestry of United States of America (USA) origin.

() antibiotic resistance poses a global health threat. Accurate identification of antibiotic resistant strains is essential for the control of infection. In the present study, our goal is to leverage the whole-genome data of to develop practical and interpretable machine learning (ML) models for comprehensive antibiotic resistance assessment. A total of 296 isolates with genome-wide data were downloaded from the Bacterial and Viral Bioinformatics Resource Center (BV-BRC) and the National Center for Biotechnology Information (NCBI) databases. By training ML models on feature sets of single nucleotide polymorphisms from SNP calling (SNPs-1), antibiotic-resistance SNP annotated by the Comprehensive Antibiotic Resistance Database (SNPs-2), gene presence or absence (GPA), we generated predictive models for four antibiotics and multidrug-resistance (MDR). Among them, the models that combined SNPs-1, SNPs-2, and GPA data demonstrated the best performance, with the eXtreme Gradient Boosting (XGBoost) consistently outperforming others. And then we utilized the SHapley Additive exPlanations (SHAP) method to interpret the ML models. Furthermore, a free web application for the MDR model was deployed to the GitHub repository (https://H.pylori/MDR/App/). Our study demonstrated the promise of employing whole-genome data in conjunction with ML algorithms to forecast antibiotic resistance. In the future, the application of this approach for predicting antibiotic resistance would hold the potential to mitigate the empiric administration.

Metabolic Associated Fatty Liver Disease (MAFLD) impacts approximately 25% of the global population. Between April 2023 and July 2023, 60 patients with MAFLD, along with 60 age, ethnicity, and sex-matched healthy controls (HCs), were enrolled from the Inner Mongolia Autonomous Region, China. Analysis of gut microbiota composition and plasma metabolic profiles was conducted using metagenome sequencing and LC-MS. LEfSe analysis identified five pivotal species: Eubacterium rectale, Dialister invisus, Pseudoruminococcus massiliensis, GGB3278 SGB4328, and Ruminococcaceae bacteria. In subgroup analysis, Eubacterium rectale tended to increase by more than 2 times and more than double in the non-obese MAFLD group, and MAFLD with moderate hepatic steatosis (HS), respectively. Plasma samples identified 172 metabolites mainly composed of fatty acid metabolites such as propionic acid and butyric acid analogues. Ruminococcaceae bacteria have a strong positive correlation with β-alanine, uric acid, and L-valine. Pseudoruminococcus massiliensis has a strong positive correlation with β-alanine. Combinations of phenomics and metabolomics yielded the highest accuracy (AUC = 0.97) in the MAFLD diagnosis. Combinations of phenomics and metagenomics yielded the highest accuracy (AUC = 0.94) in the prediction of the MAFLD HS progress. Increases in Eubacterium rectale and decreases in Dialister invisus seem to be indicative of MAFLD patients. Eubacterium rectale may predict HS degree of MAFLD and play an important role in the development of non-obese MAFLD. Eubacterium rectale can generate more propionic acid and butyric acid analogues to absorb energy and increase lipid synthesis and ultimately cause MAFLD.

, or liver fluke, causes fasciolosis in humans and livestock. Following ingestion of vegetation contaminated with encysted parasites, metacercariae, newly excysted juveniles (NEJ) excyst in the small intestine and cross the intestinal wall. After penetrating the liver, the parasite begins an intra-parenchymal migratory and feeding phase that not only drives their rapid growth and development but also causes extensive haemorrhaging and immune pathology. Studies on infection are hindered by the difficulty in accessing these microscopic juvenile parasites . Thus, a simple and scalable culture system for parasite development is needed. Here, we find that two-dimensional (2D) culture systems using cell monolayers support NEJ growth to a limited extent. By contrast, co-culture of NEJ with HepG2-derived 3D spheroids, or "mini-livers," that more closely mimic the physiology and microenvironment of liver tissue, promoted NEJ survival, growth, and development. NEJ grazed on the peripheral cells of the spheroids, and they released temporally regulated digestive cysteine proteases, FhCL3, and FhCL1/2, similar to parasites. The 3D co-culture induced development of the NEJ gut and body musculature, and stimulated the tegument to elaborate spines and a variety of surface sensory/tango/chemoreceptor papillae (termed S1, S2, and S3); these were especially pronounced around the oral and ventral suckers that sense host chemical cues and secure the parasite in tissue. HepG2 3D spheroid/parasite co-culture methodologies should accelerate investigations into the understanding of NEJ developmental biology and studies on host-parasite interactions, and streamline the search for new anti-parasite interventions.

HCO is involved in pH homoeostasis and plays a multifaceted role in human health. HCO has been recognized for its antimicrobial properties and is pivotal in bacterial antibiotic susceptibility. Notably, the interconversion between CO and HCO, facilitated by the enzyme carbonic anhydrase (CA), is crucial in tissues infected by pathogens. Studies have highlighted the antimicrobial potency of CA inhibitors, emphasizing the importance of this enzyme in this area. The potential of HCO as an antibiotic adjuvant is evident; its ability to increase virulence in pathogens such as and requires meticulous scrutiny. HCO modulates bacterial behaviours in diverse manners: it promotes O157:H7 colonization in the human gut by altering specific gene expression and, with , amplifies the effect of tobramycin on planktonic cells while promoting biofilm formation. These multifaceted effects necessitate profound mechanistic exploration before HCO can be considered a promising clinical adjuvant.

Chronic kidney and urinary tract diseases, including glomerulonephritis, nephrotic syndrome, and chronic kidney disease (CKD), present significant global health challenges. Recent studies suggest a complex interplay between infectious pathogens and immune-mediated kidney damage. This study employs Generalized Summary data-based Mendelian Randomization (GSMR) to explore causal relationships between pathogen-derived antibodies and major urinary and kidney diseases.We conducted a two-sample MR analysis using summary statistics from large-scale Genome-Wide Association Studies (GWAS) to assess associations between 46 pathogen-specific antibodies and seven urinary system diseases. We utilized robust statistical methods, including inverse variance weighting, to ascertain causal effects while controlling for potential confounders.Significant associations were identified between several pathogen-specific antibodies and disease risk. Notably, Epstein-Barr virus (EBNA-1) antibody levels were inversely associated with glomerulonephritis and nephrotic syndrome, indicating a potential protective effect. Conversely, Anti-Merkel cell polyomavirus IgG seropositivity was linked to increased risks of CKD and glomerulonephritis. Additionally, immune-mediated mechanisms were highlighted, with certain antibodies exhibiting dual roles as risk factors or protective agents.This study underscores the complex role of pathogen antibodies in the pathogenesis of kidney and urinary tract diseases, revealing significant implications for future research and potential therapeutic strategies. The findings advocate for further investigation into specific pathogen interactions with the immune system, aiming to inform targeted interventions.

toxin (PMT) is an exotoxin produced by several members of the zoonotic respiratory pathogen . The role of PMT in disrupting the mammalian respiratory barrier remains to be elucidated. In this study, we showed that inoculation of recombinantly expressed PMT increased the permeability of the respiratory epithelial barrier in mouse and respiratory cell models. This was evidenced by a decreased expression of tight junctions (ZO-1, occludin) and adherens junctions (β-catenin, E-cadherin), as well as enhanced cytoskeletal rearrangement. In mechanism, we demonstrated that PMT inoculation induced cytoplasmic Ca inflow, leading to an imbalance of cellular Ca homoeostasis and endoplasmic reticulum stress. This process further stimulated the RhoA/ROCK signalling, promoting cytoskeletal rearrangement and reducing the expression of tight junctions and adherens junctions. Notably, the T-type voltage-gated Ca channel Ca3.1 was found to participate in PMT-induced cytoplasmic Ca inflow. Knocking out Ca3.1 significantly reduced the cytotoxicity induced by PMT on swine respiratory epithelial cells and mitigated cytoplasmic Ca inflow stimulated by PMT. These findings suggest Ca3.1 contributes to PMT-induced respiratory epithelial barrier disruption.

Orf virus (ORFV) has been demonstrated to infect both goat non-immune cells, specifically goat epithelial cells, and goat blood immune cells. Our previous studies have indicated that ORFV gains entry into goat epithelial cells via clathrin-mediated endocytosis and macropinocytosis pathways. However, the pathway by which ORFV enters goat blood immune cells has not yet been elucidated. Our findings revealed a differential viral internalization pathway in ORFV-infects goat immune cells contrasting the internalization pathways in goat epithelial cells, potentially involving an antibody-related mechanism. Therefore, our hypothesis posits that ORFV gains entry into goat immune cells via the antibody-dependent enhancement (ADE) pathway. Our experimental findings confirm the presence of the ADE effect in ORFV-infected goat immune cells, mediated by Fc receptors (FcRs) as demonstrated in antibody-blocking experiments. Furthermore, the ADE effect was also observed in goat epithelial cells. Nevertheless, the ADE effect observed in goat epithelial cells was not found to be dependent on the interaction between the virus-antibody complex and Fc receptors, as demonstrated by antibody-blocking experiments. Instead, it is suggested that an alternative mechanism involving the complement factor and complement receptors (CRs) may be responsible. Overall, this research offers insights into the unique ADE pathway of ORFV infection in different cell types, offering a novel perspective on the infection and pathogenic mechanisms of ORFV.

Severe dengue often presents as shock syndrome with enhanced vascular permeability and plasma leakage into tissue spaces. studies have documented the role of Src family kinases (SFKs) and RhoA-kinases (ROCK) in dengue virus serotype 2 (DENV2)-induced endothelial permeability. Here, we show that the FDA-approved SFK inhibitors Bosutinib, Vandetanib and Ponatinib, as well as the ROCK inhibitors, Netarsudil and Ripasudil significantly inhibit DENV2-induced endothelial permeability. In cultured telomerase immortalized human microvascular endothelial cells (HMEC-1), treatment with these inhibitors reduced the phosphorylation of VE-Cadherin, Src and myosin light chain 2 (MLC2) proteins that were upregulated during DENV2 infection. It also prevented the loss of VE-Cadherin from the inter-endothelial cell junctions induced by viral infection. In studies using DENV2-infected AG129 IFN receptor-α/β/γ deficient mice, ponatinib, when administered 24 h post-infection onwards, demonstrated significant benefits in improving body weight, clinical outcomes, and survival rates. While all virus-infected, untreated mice died by day-10 post-infection, 80% of the ponatinib-treated mice survived, and approximately 60% were still alive at the end of the 15-day observation period. The treatment also significantly reduced disease severity factors such as vascular leakage, thrombocytopenia; mRNA transcript levels of proinflammatory cytokines such as IL-1β and TNF-α; and restored liver function. Comparable effects were observed even when ponatinib treatment was initiated after symptom onset. The results highlight ponatinib as an effective therapeutic option in severe dengue; and also a similar potential for other FDA- approved SFK and ROCK inhibitors.

The high-osmolarity glycerol (HOG) signalling pathway, comprising Ste11/Ssk2/Ssk22 (MAPKKK), Pbs2 (MAPKK), and Hog1 (MAPK), is an important and conserved pathway in fungi. However, the functions and downstream regulatory factors of Hog1 in nematode-trapping (NT) fungi remain poorly understood. Here, three proteins (AoNmd5, AoPyp1, and AoPtp) interacting with Hog1 were screened in a representative NT fungus using yeast screening library and verified using yeast two-hybrid (Y2H) assay. The function of AoNmd5 was furtherly characterized by phenotypic comparison, staining technique, and multi-omics analyses. AoNmd5 was essential for vegetative growth, conidial development, trap morphogenesis, and nematode predation ability. In addition, AoNmd5 played crucial roles in endocytosis, lipid metabolism, reactive oxygen species, stress response, autophagy, and other metabolic processes. Furthermore, we constructed an AoNmd5 interaction network based on transcriptomic analysis and Y2H, revealing its significant role in the respiratory chain and redox processes as well as its interaction with the small GTPase Ran1, which mediates Hog1 nucleocytoplasmic shuttling. These findings suggest that the Hog1-Nmd5 signalling pathway has pleiotropic roles in . This study deepens our understanding of the HOG pathway and its interaction with importins in NT fungi.

Honey bees are important pollinators in both agriculture and ecosystems, and their health is essential for sustainable human development. Although only two bacteria, and , have been identified as bacterial pathogens in honey bee brood for over 100 years, we found three additional strains ( sp. J27TS7, J34TS1, and J46TS7) in honey that harmed honey bee brood development. In particular, sp. J27TS7 was highly virulent in bee larvae (the median lethal dose [LD] = 12.7 spores/larva) and was comparable to (LD = 2.3-11.5 spores/larva). J34TS1 showed the second-highest virulence (LD = 45.9 spores/larva), and J46TS7 was the least virulent (LD = 469.0 spores/larva). However, was most frequently detected in Japanese honey among the three species, with the highest concentration being 1.8 × 10 spores/mL honey, suggesting its wide distribution in Japanese apiaries. The novel pathogenic species were categorized into the fast killer ( sp. J27TS7), medium-fast killer (), and slow killer () like strains in terms of the time to kill infected brood; however, histopathological and genome analyses indicated that their pathogenic mechanisms were different from those of strains. Moreover, showed differences in virulence depending on the lineage of the strain. These findings represent the first discovery of honey bee brood pathogens in more than 100 years and indicate the need to look beyond known pathogens for a comprehensive understanding of honey bee diseases.

Canine distemper virus (CDV) is a member of the genus with a worldwide distribution that causes fatal diseases in canids and marine mammals. In recent years, CDV has demonstrated the remarkable ability of pathogens to cross species barriers. The natural host range of CDV has expanded from Canidae to Primates, presumably attributed to ecological shifts and the emergence of viral variants. Therefore, it is important to investigate whether CDV can infect humans by adapting to the human signalling lymphocyte activation molecule (hSLAM) receptor to cross the species barrier. Through successive passaging and plaque cloning of a CDV wild-type strain (5804PeH) in Vero cells expressing hSLAM (Vero-hSLAM), we obtained an hSLAM adaptive strain, 5804PeH-VhS. The adapted CDV strain exhibited a D540G mutation within the receptor-binding domain (RBD) of the haemagglutinin (H) protein. The H mutation has enhanced cell-cell fusion activity in Vero-hSLAM cells. This adaptation allowed the CDV strain to infect human peripheral blood mononuclear cells (PBMCs), particularly T lymphocytes and inhibited lymphocyte proliferation. Additionally, this strain could replicate in the lymphoid tissues of transgenic mice that express the hSLAM receptor, causing viraemia. However, the adapted strain did not spread to the epithelial cells or the central nervous system of the mice. While this adaptation indicates a potential risk, there is no definitive evidence that the virus can spread among humans.

, a zoonotic pathogen, is commonly found as a commensal bacterium in the respiratory tracts of pigs. Under specific conditions, it becomes invasive and enters the blood, causing severe systemic infections. For , effective acquisition of carbon sources in different host niches is necessary for its survival. However, as of now, our understanding of the metabolism of within the host is highly restricted. Pyruvate formate lyase (PFL) plays a crucial role in bacterial survival of in glucose-limited and hypoxic host tissues. Here, we investigated the physiological and metabolic functions of PFL PflB in and elucidated its pivotal role in regulating virulence within the mucosal and blood niches. We demonstrate that PflB is a key enzyme for to support mixed-acid fermentation under glucose-limited and hypoxic conditions. Additionally, PflB is involved in regulating morphology and stress tolerance, and its regulation of capsular polysaccharide content depends on dynamic carbon availability. We also found that PflB is associated with the capacity of to cause bacteremia and persist in the upper respiratory tract to induce persistent infection. Our results provide highly persuasive evidence for the relationship between metabolic regulation and the virulence of .

is a prevalent opportunistic pathogen responsible for a wide range of infections in livestock and wildlife, such as in cattle, pigs, European bison and forest musk deer. Much of the successful infection of relies on its virulence factors, including pyolysin as well as adhesion factors. The swift rise of bacterial resistance has highlighted the urgent need for developing new therapeutic strategies. Currently, virulence factor-mediated vaccine development and other therapeutic approaches are widely regarded as the primary interventions for addressing diseases associated with this pathogen. This review examines the broader virulence potential of , focusing on haemolysin, host cell adhesion proteins, the prevalence of antibiotic resistance, and the development of vaccines mediated by virulence factors. Additionally, it discusses current and future approaches aimed at improving therapeutic interventions.

Chemosensory protein 16 was identified in the hemolymph of as a protein with an amount increasing after oral infection with 10^3 CFU of , and decreasing after infection with a higher dose (10^5 CFU) of bacteria. The expression of the CSP16 gene occurred in the fat body and in the gut and correlated with changes in the protein level in the hemolymph. The CSP16 protein inhibited growth in the concentration range from 0.15 to 6 nM. Additionally, the CSP16 protein showed bactericidal activity against , and in the range of 2-18 μM, but only in the presence of protease inhibitors, otherwise it was degraded by extracellular proteases secreted by . We demonstrated that the bactericidal activity of CSP16 was related to its ability to perforate bacterial cellular membranes in a dose-dependent manner. The antimicrobial properties of this protein were also confirmed with the use of Atomic Force Microscopy, which showed significant changes in the topology of different bacterial cell surfaces. Finally, when CSP16 was injected into larvae one hour after infection with , more survivors were observed at particular time-points. Taking into account its immune properties and putative ability to bind bacteria-derived compounds, the possible function of CSP16 in the host-pathogen interaction is discussed.

The mammalian intestine is a major site of colonization and a starting point of severe infections by . Inflammatory bowel disease (IBD) is an inflammatory disorder of the gut, and host-derived nitrate in IBD confers a luminal growth advantage upon and through nitrate respiration in the inflamed gut. However, the impact of nitrate on the growth and pathogenicity of in this microenvironment is poorly understood. In this study, we used oral administration of dextran sodium sulphate to induce IBD in mouse models. We then analysed the colonization levels of wild-type (WT), the nitrate reductase gene mutant strains (Δ, Δ and ΔΔ), and the molybdate uptake gene mutant strain (Δ) in the inflamed intestinal tract. Results showed that the growth, intestinal colonization, and extraintestinal dissemination of were increased in the intestines of dextran sulphate sodium (DSS)-treated mice. Nitrate in the inflamed bowel conferred a growth advantage to through nitrate respiration. The molybdate transport protein ModA regulated nitrate reductase activity to increase the growth, intestinal colonization, and extraintestinal dissemination of . Tungstate will be a promising antibacterial agent to tackle infections in IBD patients.

The fungus , which impacts rice spikes, causes rice false smut (RFS), a significant prevalent disease in rice cultivation regions globally. Cytochrome genes are known to be involved in secondary metabolism and pathogenesis in various species, but studies on genes in are limited. In this research, a family gene, , was found up-regulated during invasion stage of . Observation of fluorescence indicated that UvCYP503-GFP is situated within cytoplasm of hyphae. Disruption of led to decreased hyphal development, conidiation, and pathogenicity. Additional RNA-seq assay revealed that affects the transcript of genes associated with pathogenicity, various stress responses, and other genes. In alignment with RNA-seq results, compared with wild-type, Δ mutants showed increased sensitivity to cell wall stresses, but reduced sensitivity to osmotic and hyperosmotic stressors. Moreover, Δ mutants exhibited decreased sensitivity to the fungicides difenoconazole and tebuconazole. This study represents a phenome-based functional analysis of a gene in and provides valuable genetic resources for further research in filamentous fungi and other plant pathogens.

Viral myocarditis (VM) is an inflammatory disease posing a serious threat to public health, with various viral pathogens contributing to its pathogenesis. Coxsackievirus B3 (CVB3) is the most frequently implicated causative agent and has been extensively studied because of its high prevalence and severity. No specific therapeutic interventions for VM exist, and vaccine development has encountered substantial challenges. Therefore, we aimed to develop a novel CVB3 mucosal vaccine as a preventive strategy against VM. Gram-positive enhancer matrice (GEM) particles serve as innovative mucosal vaccine adjuvants and antigen delivery systems that enhance antigen immunogenicity by facilitating effective mucosal immune responses. In this study, GEM particle display technology was used to develop two novel CVB3 vaccines: (1) a GEM particle-based vaccine displaying the CVB3 capsid protein VP1 via a PA anchor protein (GEM-PA-VP1), and (2) a GEM particle-based vaccine displaying VP1 via the FcSP peptide (GEM-Fc-VP1). Both, GEM-PA-VP1 and GEM-Fc-VP1 elicited significantly elevated levels of specific IgG, IgG1, IgG2a, SigA and neutralizing antibodies in a mouse model, along with enhanced secretion of Th1- and Th2-associated cytokines, compared to controls. Notably, GEM-Fc-VP1 demonstrated superior immunogenicity compared with that of GEM-PA-VP1, evidenced by higher antibody titres and cytokine responses. In challenge protection experiments, both vaccines significantly improved survival rates, reduced myocardial enzyme levels, and decreased inflammatory cell infiltration in myocardial tissue, with GEM-Fc-VP1 exhibiting greater efficacy. These findings establish a foundation for the development of a safe and effective CVB3 candidate vaccine and provide novel insights into the potential of peptide-mediated subunit vaccine approaches.