Search Results (1,384)

Oral, live-attenuated rotavirus vaccines (ORVs) have significantly reduced severe diarrheal disease and deaths; however, vaccine performance in low- and middle-income countries (LMICs) remains lower than in high-income countries. With increasing global use, real-world data now provide important insights into strategies to improve vaccine impact in these settings. This review summarizes current evidence on the effectiveness and impact of ORVs in LMICs, with a focus on approaches to optimize performance, including alternative dosing schedules, booster doses in late infancy, and mixed vaccine regimens. Evidence on vaccine interchangeability offers practical solutions for countries facing supply disruptions and financial constraints. The review also highlights the growing importance of economic sustainability, particularly in the context of transitions from external funding support. Despite lower efficacy, ORVs continue to provide substantial public health benefits in LMICs. Sustained impact will depend on strengthening programme delivery, ensuring stable vaccine supply, and advancing next-generation vaccines. Continued evaluation of effectiveness, cost, and implementation strategies will be critical to maximizing long-term benefits in high-burden settings. Full article

Objective: It is commonly believed in the public health community that the mRNA vaccines from Pfizer and Moderna were broadly similar in preventing COVID-19 mortality, with perhaps a small advantage for Moderna. We revisit this question, using a research design that controls for healthy-vaccinee bias. Methods: We studied deaths among persons aged 15 years or greater from January 2021 through December 2022 in Indiana, linked to vaccination records, using an outcome measure, namely COVID-19 Excess Mortality Percentage (CEMP), which uses non-COVID natural mortality to control for HVB. Results: We find both large healthy-vaccinee bias for both vaccines, and larger bias for Pfizervaccinees: Pfizer recipients were healthier and thus would have faced lower COVID-19 mortality without vaccination. For ages 60+, we find almost 50% higher COVID-19 remaining mortality risk (RMR) for Pfizer vaccinees receiving primary vaccination over a “core period” from 2Q2021 through 1Q2022: the average Pfizer/Moderna RMR ratio was 1.49. For ages 15–59, this ratio was 1.25, but was not statistically significant. For booster dose for ages 60+, the average Pfizer/Moderna RMR ratio was 1.34. For 2Q2022 through 4Q2022, after the major Omicron wave, we find much lower vaccine effectiveness and no significant Pfizer–Moderna difference. Conclusions: During the period with both good vaccine availability and high COVID-19 mortality (2Q-2021–1Q2022), the Moderna vaccine was associated with lower COVID-19 mortality, relative to the Pfizer vaccine. A possible explanation is the higher Moderna dose (100 μg vs. 30 for Pfizer). After that period, a high percentage of the population had natural immunity from prior COVID-19 infection; we find only small mortality gains from vaccination, with no significant Pfizer–Moderna difference. Policy Implications: Controlling for healthy-vaccinee bias, Moderna was associated with lower COVID-19 mortality, relative to Pfizer. This difference was not known in real-time due to the failure of the available studies to address healthy-vaccinee bias. Controlling for such bias is vital when making vaccination policy. Full article

Background: Staphylococcus aureus (SA) causes septicemia and arthritis, eventually resulting in substantial economic losses and health hazards in poultry. Lethal electron beam (eBeam) treatment kills bacteria while preserving surface epitopes. Methods: This study compared broiler chicken immune responses (local and systemic) after eBeam-killed (eB) or formalin-killed (FK) SA injections. The vaccine (sham) control was endotoxin-free PBS. This study contained six treatments (trt) with five chickens/trt, with vaccine trts divided into two groups, A and B. Group A received in ovo vaccine/sham treatments (phase 1) initially. At 34 d of age, the pulps of growing feathers (GFs) received intradermal (i.d.) injections of the respective trt to elicit booster and primary responses in groups A and B, respectively. Blood was collected to analyze leukocyte populations and plasma SA-specific antibody levels. Additionally, in phase 2, GFs were collected to assess leukocyte presence in GF pulps. Two-way ANOVA was conducted to test the effects of treatment, time, and their interactions, followed by Tukey’s HSD tests at p < 0.05 for statistical significance. Results: Early in phase 1, the eB group had increased SA-specific IgM, IgA, and total lymphocyte concentrations compared with FK. In phase 2, FK and eB i.d. vaccines increased total lymphocyte and T cell proportions in GF pulps compared with sham. In blood, the recall eB vaccination increased monocytes, B cells, CD8+ T cells, and total lymphocytes, while the recall FK vaccination increased heterophils (p < 0.05). Conclusions: Improved early protection at mucosal surfaces may be reflected by higher plasma levels of SA-specific IgM and IgA following in ovo eBeam-killed-SA vaccination. Additionally, both eB-SA and FK-SA vaccines stimulated robust local inflammatory responses dominated by lymphocytes in GF pulps. Full article

Bovine coronavirus (BCoV) is a major cause of respiratory and enteric diseases in cattle, resulting in substantial economic losses to the global cattle industry. Next-generation epitope-focused vaccines require conserved neutralizing determinants to achieve broad protective efficacy across circulating variants. Here, we applied a reverse-vaccinology and immunoinformatics-guided strategy to identify conserved linear B-cell epitope candidates on the BCoV spike protein. Using ABCPred (16 aa; threshold score 0.51), 139 putative linear B-cell epitopes were predicted, and 10 candidates (B1–B10) were selected based on antigenicity, allergenicity/toxicity filters, and Shannon-entropy-based conservation (≥95%). Each candidate was displayed on Helicobacter pylori ferritin and TEM confirmed the formation of self-assembled nanocages. BALB/c mice (n = 6 per group) were subcutaneously immunized with 50 μg of each epitope–ferritin fusion protein emulsified 1:1 with Montanide ISA 201 in a prime–boost regimen (at a 14-day interval), and sera were collected 14 days after the booster immunization. All 10 constructs induced BCoV-specific binding IgG (endpoint titers ranging from 1:2667 to 1:11,733), but only B1, B2, B4, B6, B7, and B8 elicited detectable in vitro neutralizing activity against both representative strains, whereas B3, B5, B9, and B10 remained below the detection limit (<1:8). Neutralizing titers were 1:24–1:99 for the enteric strain XHD4 and 1:27–1:88 for the respiratory strain HXD1, with no significant difference observed between the two strains (p > 0.05). Overall, these findings identify six conserved B-cell epitope candidates capable of eliciting neutralizing antibody responses against both representative enteric and respiratory field strains, providing experimental evidence to support epitope-focused BCoV vaccine design. Full article

Background/Objectives: Pseudorabies virus (PRV) variant strains have caused widespread outbreaks in China since 2011, and currently available vaccines provide suboptimal protection. Glycoprotein D (gD), the principal target of virus-neutralizing antibodies, represents a promising antigen for subunit vaccine development. However, CHO cell-based production systems suitable for large-scale manufacturing remain insufficiently explored. This study aimed to develop a potentially scalable CHO cell-derived PRV gD subunit vaccine and evaluate its immunogenicity and protective efficacy in mice. Methods: A stable Chinese hamster ovary (CHO) suspension cell line secreting the extracellular domain of PRV gD was established through signal peptide optimization and stepwise serum-free adaptation. The recombinant gD protein was purified using Ni²⁺- Sepharose High-Performance affinity chromatography and subsequently formulated with MONTANIDE ISA 206 adjuvant. Immunogenicity and protective efficacy were assessed in BALB/c mice through serological analysis, neutralization assays, lethal challenge experiments, and quantitative PCR. Results: The gD subunit vaccine induced rapid seroconversion of gD-specific IgG antibodies as early as 7 days post immunization and exhibited a strong booster effect, maintaining high antibody levels. Neutralizing antibodies were first detected at 14 days and increased significantly after booster immunization, with titers markedly exceeding those induced by a commercial inactivated PRV vaccine at 42 days (p = 0.001). Following lethal challenge with 10⁴ TCID₅₀ of the highly virulent PRV-TJ variant strain, vaccinated mice achieved 100% survival without clinical signs. Viral genome copy numbers in the brain and spinal cord were reduced by approximately 3.3 to 4.4 log₁₀ relative to the PBS control group. Conclusions: The CHO cell-derived PRV gD subunit vaccine elicits robust humoral immune responses and provides complete protection against lethal PRV variant challenge in mice. These findings support its further evaluation in the natural swine host toward the development of a safe and scalable subunit vaccine for pseudorabies control. Full article

Background: Pharmacovigilance systems have flagged possible associations between mRNA COVID-19 vaccination and systemic autoimmune inflammatory rheumatic diseases (AIRDs), generating uncertainty for rheumatologists counselling patients. As the first population-scale deployment of an mRNA vaccine platform, COVID-19 vaccination provides a unique setting to examine whether autoimmune safety signals detected in spontaneous reporting systems correspond to measurable disease risk. We synthesised pharmacovigilance and population-based evidence on incident EULAR-defined systemic AIRDs after mRNA COVID-19 vaccination and assessed whether disproportionality signals were corroborated by analytical studies. Methods: We conducted a PRISMA 2020-compliant systematic review searching MEDLINE, Web of Science, Scopus, Embase, and the Cochrane Library from 2019 to April 2026, supplemented by medRxiv and trial registries. Eligible studies included pharmacovigilance disproportionality analyses and analytical studies, including cohorts and randomised controlled trials, evaluating BNT162b2 or mRNA-1273 in adults without known pre-existing autoimmune disease. Risk of bias was assessed using READUS-PV, ROBINS-I, and RoB 2. Meta-analysis was not performed because of substantial heterogeneity. Results: Fourteen studies were included: seven pharmacovigilance studies and seven analytical studies. Disproportionality analyses suggested increased reporting of selected AIRDs, most consistently polymyalgia rheumatica and giant cell arteritis, mainly when all other adverse-event reports served as comparators. These signals were largely neutral when influenza vaccines were the reference. Across analytical studies, associations were inconsistent; modest increases in systemic lupus erythematosus appeared only in selected analyses. Long-term evidence was scarce: only four studies, from three countries (South Korea, Israel, and Norway), followed participants for up to approximately one year, and three of these reported at least one positive association—systemic lupus erythematosus, post-booster rheumatoid arthritis, and polymyalgia rheumatica in older adults—whereas studies restricted to risk windows of three months or less reported no increase. Conclusions: The available evidence does not indicate a consistent increase in incident systemic AIRDs after mRNA COVID-19 vaccination. Although pharmacovigilance studies identified comparator-dependent signals for selected diseases, particularly polymyalgia rheumatica and giant cell arteritis, these findings were generally not confirmed in comparative population-based studies and should be considered hypothesis-generating. Delayed-onset disease remains poorly characterised, and studies with at least one year of follow-up are needed. Full article

The largemouth bass (Micropterus salmoides) is persistently threatened by Micropterus salmoides rhabdovirus (MSRV) and largemouth bass ranavirus (LMBV), highlighting the need for combined immunization strategies. Here, a bivalent DNA vaccine pEGFP-G-MCP was constructed by concatenating the highly immunogenic fragments of MSRV glycoprotein (MSRV-G) and LMBV main capsid protein (LMBV-MCP) using a flexible linker peptide, and its immunogenicity and protective effect were systematically evaluated. Fish were immunized by intramuscular injection of 200 ng of plasmid on day 1, followed by a booster immunization on day 8. Blood and tissues were collected at 10, 20, and 30 d post-last immunization. The results showed that pEGFP-G-MCP immunization induced G- and MCP-specific antibodies and upregulated the expression of immunity-related genes. The bivalent vaccine achieved a relative percent survival (RPS) of 93.3% against MSRV and 90.0% against LMBV, which were higher than those of the monovalent vaccine groups. Additionally, the bivalent vaccine reduced viral mRNA and alleviated pathological damage in MSRV- or LMBV-infected fish. These results indicate that pEGFP-G-MCP confers immune protection against MSRV and LMBV, providing a foundation for combined prevention and control of MSRV and LMBV. Full article

Background: Vaccine hesitancy is a global challenge affecting workplaces and academic institutions. Despite efforts, low vaccination rates persist in South Africa, increasing the risks associated with pathogen exposure. This study assessed the knowledge, attitudes, and perceptions (KAPs) of healthcare workers (HCWs) and students at a Nursing College regarding voluntary and mandatory COVID-19 vaccination. Methods: A quantitative cross-sectional study was conducted using a structured, self-administered electronic questionnaire. All eligible nursing students and staff at the participating Nursing College were invited to participate. Of the 504 eligible individuals, 372 completed the questionnaire, corresponding to a response rate of 73.8%. Data were analyzed using IBM SPSS Statistics version 30.0. Logistic regression and Mann–Whitney U tests were used to assess associations and differences between groups, with statistical significance set at p < 0.05. Results: Most respondents were female (302, 81.0%) and single (252, 67.9%). The age group 26 to 40 was most prominent (144, 38.8%). Only 42.2% of respondents (n = 157) were fully immunized with no boosters. Student nurses dominated (344, 92.5%), with 226 (60.9%) having no COVID-19 infections and 198 (72.0%) voluntarily receiving the vaccine. Occupation (p = 0.030) and age group (p = 0.027) were independently associated with reporting voluntary COVID-19 vaccination. Age was also associated with respondents’ attitudes and perceptions of the COVID-19 vaccine (p < 0.001) as well as their understanding of the vaccine (p = 0.002). Conclusions: Occupation and age group were independently associated with the reported basis on which COVID-19 vaccination was received (voluntary versus mandatory). Targeted communication and educational strategies addressing these factors may improve vaccine acceptance and preparedness for future outbreaks. Full article

Background: Individuals with autoimmune disease, inflammatory bowel disease (IBD), and transplants face a higher risk of severe COVID-19 and breakthrough hospitalizations. It is essential to understand the magnitude and durability of vaccine-induced humoral immune response in these populations. Methods: We evaluated SARS-CoV-2 spike IgG antibody levels and avidity in autoimmune, IBD, transplant, and healthy cohorts following multiple COVID-19 mRNA vaccine doses. Serum samples were collected approximately 1 month (8–52 days) and 6 months (158–202 days) post-vaccination. Antibody levels and avidity were measured using validated ELISA and chaotropic-based avidity assays. Results: Individuals with IBD and individuals with autoimmune disease, especially systemic autoimmune disease, exhibited lower antibody levels and avidity compared with healthy individuals at certain doses and time points. Transplant recipients demonstrated substantial impairments in both antibody levels and avidity, with avidity reduced across all doses and time points. Significantly lower avidity levels were observed in transplant recipients, suggesting challenges in developing or maintaining antibody quality. For example, geometric mean anti-spike IgG levels were substantially lower in transplant recipients than in healthy individuals at both 1 month (635 vs. 7685 BAU/mL; p < 0.0001) and 6 months (1146 vs. 3277 BAU/mL; p = 0.0057) post third dose. Similarly, transplant recipients had lower antibody avidity than healthy individuals after the third dose, with geometric mean AI80 values of 4.5 M vs. 5.5 M at 1 month (p < 0.0001) and 4.6 M vs. 5.4 M at 6 months (p < 0.0001). Age, sex, and vaccine manufacturer may further influence humoral immune responses in the transplant cohort. Conclusions: Vaccine-induced humoral immunity varies across autoimmune, IBD, and transplant cohorts, with the most persistent impairment observed in transplant recipients across vaccine dose groups and at both post-vaccination time points. These findings reveal immune response patterns that may guide future studies evaluating vaccination schedules, immune monitoring, and clinical outcomes in these populations. Full article

Background: Despite the increased infection-related morbidity of patients with chronic kidney disease (CKD), their vaccination coverage is low. Cocooning strategy has not been adequately researched in this group. Our aim was to evaluate vaccination coverage of CKD pediatric and adult patients and their families. Methods: In this prospective, single-center study, we recorded the vaccination coverage of CKD pediatric and adult patients and their families who attend a tertiary University Hospital. Vaccination rates were calculated according to the national vaccination program. Vaccination of pediatric CKD patients was compared to a control group of healthy children. Results: The vaccination rate of 63 hemodialysis patients was low for influenza (61.9%), COVID-19 (79.4% primary, 3.2% booster doses), RSV (50%), herpes zoster (42.4%), hepatitis B (41.3%), Streptococcus pneumoniae (25.4%), tetanus–pertussis (3.2%), and HPV (0%). Their 30 underaged relatives had low vaccination rates for influenza (13.3%), DTaP/Tdap (80%), and Men B (50%), with adequate vaccination for other pathogens. The 53 children with CKD were poorly vaccinated for influenza (20.8%), COVID-19 (0%), Streptococcus pneumoniae (15.4%), Men B (30.2%), DTaP/Tdap (67.9%), MMR (78.8%) and VZV (86.5%), but their vaccination rates for influenza were significantly higher compared to controls. Their 108 adult relatives were inadequately vaccinated for influenza (27.8%), COVID-19 (1.9%), and Tdap (1.9%). Their 35 underaged relatives were poorly vaccinated for influenza (22.9%) and DTaP (80%). Conclusions: Vaccination coverage of CKD patients is suboptimal for vaccines particularly important for their condition and their family members are insufficiently informed about potential contribution to their protection by cocooning strategy. Full article

Porcine epidemic diarrhea virus (PEDV) is a major enteric coronavirus that causes severe economic losses to the global swine industry. Current vaccines suffer from weak immunogenicity, insufficient mucosal immunity, and a short duration of protection. Nanoparticle delivery systems have emerged as a promising strategy for next-generation vaccine development. In the present study, we constructed an S1-Ferritin nanoparticle vaccine using the SpyTag-SpyCatcher modular conjugation system. The morphology, particle size, and uniformity of the nanoparticle vaccine were systematically characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). After two immunizations, the S-Ferritin nanoparticle vaccine elicited potent humoral and cellular immune responses in both mice and piglets. In piglets, at 2 weeks post booster vaccination, PEDV-specific serum IgG endpoint titers peaked at 1:2560, virus-neutralizing antibody titers reached 1:256 against the JS-2/2015 strain, and serum IFN-γ levels reached 274 pg/mL. All these immunological indicators were significantly higher than those observed in the PEDV-inactivated whole-virus vaccine group. Furthermore, challenge tests showed that the S1-Ferritin nanoparticle vaccine provided complete protection against PEDV infection and significantly reduced the severity of diarrhea and intestinal damage in piglets after challenge. These findings suggest that the S1-Ferritin nanoparticle vaccine constitutes a promising candidate against PEDV infection, though our study is only a preliminary step and subsequent field trials in pigs are still required. Full article

Veterinarians are routinely exposed to occupational infectious hazards at the human–animal interface, making vaccination an important preventive measure within a One Health framework. However, evidence on vaccination coverage and immunization practices in this professional group remains limited. This study assessed self-reported vaccination coverage, attitudes toward vaccination, perceived barriers, and training needs among registered veterinarians in Málaga, Spain. A cross-sectional anonymous survey was distributed to all actively practicing veterinarians registered with the Official Veterinary Association of Málaga. The questionnaire collected sociodemographic and professional information, self-reported vaccination status, attitudes toward vaccination, and perceived training needs. A total of 164 veterinarians participated (response rate: 15.2%). Given the voluntary nature of participation, findings should be interpreted as descriptive. Vaccination coverage was low for several vaccines relevant to occupational or adult immunization, including seasonal influenza (21.3%), rabies (23.2%), and tetanus–diphtheria–pertussis boosters (47.3%). Nearly half of respondents (49.1%) were uncertain about their hepatitis B vaccination status. In contrast, COVID-19 vaccination uptake was high (96.9% received at least one dose). Although most participants considered vaccination highly important, self-reported coverage remained limited for several recommended vaccines. Qualitative responses identified barriers related to access to vaccination, information, and the absence of structured occupational vaccination programmes. Among the surveyed veterinarians in Málaga, these findings support the need for clearer occupational vaccination recommendations, improved access to immunization services, and enhanced vaccinology training. Such measures could strengthen occupational health protection and support One Health prevention strategies at the human–animal interface. Full article

Background: Adenoviral vectors are widely used in vaccine development; however, pre-existing immunity to common human adenovirus serotypes can limit their effectiveness. Canine adenovirus type 2 (CAV-2) is a non-human adenovirus with low seroprevalence in humans, making it a suitable alternative vector. Despite its promise, comprehensive preclinical safety data for CAV-2 vector-based vaccine platforms remain limited. In this study, we evaluated the safety profile of a replication-defective CAV-2 vector expressing the Omicron BA.4 SARS-CoV-2 spike immunogen in BALB/c mouse models. Methods: The SARS-CoV-2 CAV-2 vector-based vaccine was expressed and propagated in AD293 cells. The mice received intramuscular prime-boost immunisations with low (1 × 10⁶ PFU), moderate (0.5 × 10¹⁰ PFU), or high (1 × 10¹⁰ PFU) vaccine doses, alongside empty CAV-2 vector and physiological buffer control groups, and were monitored longitudinally up to Day 72. The mice were clinically assessed at days 0, 7, 21, 42, and 72 for any deviations from normal conditions in comparison to the control groups. Biochemical analyses were performed to evaluate liver and kidney function, as well as any tissue injury due to the vaccine candidate. Hematological parameters were assessed by conducting complete blood counts. Body temperature and weight changes were also monitored as an indicator of systemic toxicity. Results: The biochemical and haematological parameters remained within physiological reference ranges across all dose groups and timepoints, with no dose-related deviations, indicating that the vaccine candidate did not show evidence of hepatotoxicity, nephrotoxicity, tissue or haematological toxicity. Body temperatures remained within normal physiological ranges following both prime and booster immunisations, and body weights increased normally across all groups as the animals grew throughout the study period without any abnormal weight gain or loss. Conclusions: These results suggest that the replication-defective CAV-2-vectored SARS-CoV-2 vaccine candidate was well tolerated and did not demonstrate evidence of systemic toxicity under the conditions tested. These findings demonstrate that the CAV-2 vector exhibits a favourable safety profile in murine models when used as a vaccine delivery platform, supporting its translational potential as an alternative adenoviral vector-based vaccine platform. Further studies incorporating additional safety endpoints, including histopathological, vector persistence and shedding evaluation, are warranted to support continued development of the platform. Full article

Background: Buffaloes have a significant share in the agricultural economies of many developing countries; however, the industry is not well-established because of the low reproductive efficiency of buffaloes. In this study, we designed a novel strategy to enhance buffalo reproductive efficiency and examined the significance of intramuscular AMH-INH-RFRP DNA vaccine immunization on reproductive performance. Methods: Allocated into four groups (n = 30/group), female buffaloes (aged 5–8 years, n = 120) were randomly enrolled in the study. Treatment groups received 10 mL of AMH-INH-RFRP DNA vaccine intramuscularly, once daily for three consecutive days, at the following concentrations: T1—3 × 10⁸ CFU/mL, T2—3 × 10⁹ CFU/mL, and T3—3 × 10¹⁰ CFU/mL. All vaccinated groups received a booster vaccination two weeks later, while the control group received 10 mL of phosphate-buffered saline (PBS) intramuscularly on the same schedule. Serum antibody titers against anti-Müllerian hormone (AMH), inhibin (INH), and RF-amide-related peptide (RFRP) were quantified using indirect ELISA on days 14 (post-primary immunization) and 28 (post-booster immunization). Results: Following primary immunization, antibody titers against AMH, INH, and RFRP were significantly higher in the T3 group than in the T1 group (p < 0.05), and booster immunization further increased antibody positivity rates. Serum concentrations of IL-4, IFN-γ, and E₂ were significantly elevated in the T2 and T3 groups compared to the control group (p < 0.05), while P₄ levels showed no significant differences among groups. Ultrasonography revealed that the ovulatory follicle diameter and dominant follicle growth rate were significantly increased in the T2 and T3 groups compared to the control and T1 groups (p < 0.05). The estrus rate was significantly higher in the T3 group than in the control group (76.67% vs. 40.00%, p < 0.05), and ovulation rates were significantly higher in the T2 (83.33%) and T3 (86.67%) groups compared to the control group (56.67%) (p < 0.05). However, conception rates did not differ significantly between vaccinated and control groups in the primary dose comparison (p > 0.05). In exploratory post hoc analyses, antibody-positive (Ab+) buffaloes exhibited higher E₂ concentrations, larger ovulatory follicle diameters, accelerated dominant follicle growth rates, and higher estrus and ovulation rates compared to antibody-negative (Ab−) buffaloes. Furthermore, gestating buffaloes were monitored until calving, revealing no significant effect of the vaccine on newborn calf weight or body size. Conclusions: In total, intramuscular immunization with the AMH-INH-RFRP DNA vaccine, particularly at the highest dose, enhances reproductive performance in buffaloes by stimulating E₂ secretion and follicular development. Full article

Dengue fever, a rapidly spreading mosquito-borne viral disease, poses a major global health issue, particularly in tropical and subtropical regions. The absence of a universally effective vaccine against all four dengue virus serotypes necessitates continued exploration of novel vaccine strategies. This study evaluated a PrM-derived multiple antigenic peptide (MAP) vaccine candidate in BALB/c mice. Molecular docking analyses predicted favourable interactions of the selected PrM-derived B-cell epitope with both human (human BCR Fab region PDB ID: 5IFH) and mouse (mouse BCR PDB ID: 8EMA) B-cell receptors, supporting its antigenic potential. ChemSketch was used to develop the PrM–MAP vaccine construct, which was then commercially synthesized. Mice received single, double, and triple booster doses of the vaccine, while control groups received either PBS or adjuvant alone. Humoral immune responses were measured by ELISA, and safety was evaluated through in vitro cytotoxicity assays on HEK293 cells and the in vivo histopathological examination of major organs. Mice immunized with a single booster dose produced significant levels of PrM-specific antibodies compared to subsequent booster doses, suggesting variability in the humoral response elicited. Optimal coating concentrations in ELISA demonstrated the importance of antigen concentrations in the sensitivity of the assay. The safety evaluation indicated high cell viability, up to 150 µg/mL of PrM–MAP, in HEK293 cells, and an absence of significant histopathological damage in treated animals. The PrM–MAP vaccine construct elicited a detectable humoral immune response while demonstrating a high safety profile in vitro and in vivo. These findings indicate the potential of MAP-based vaccine platforms as promising candidates for further development against the dengue virus. Full article