Vaccines

Introduction: Recommendations from physicians and peers play a crucial role in promoting vaccination. This study evaluated differences in recommendations to others regarding four vaccines with varying efficacy (seasonal influenza, measles, human papillomavirus [HPV], and coronavirus disease 2019 [COVID-19]) between physicians and the general public and examined the impact of birth-year-based vaccination policy changes on these recommendations. Methods: This cross-sectional study was conducted in February 2026 among 492 physicians and 5,252 members of the general public in Japan. Consistency in recommendations across the four vaccines was assessed using the intraclass correlation coefficient (ICC[3,1]), and group differences were examined using a two-way mixed-design analysis of covariance. Multilevel regression discontinuity analyses were performed to evaluate the effects of birth-year-based vaccination policy. Results: Physicians showed significantly stronger recommendations to others than the general public, and their recommendation patterns generally reflected vaccine efficacy. However, physicians showed lower consistency across vaccine types than the general public (ICC[3,1]), driven primarily by heterogeneity in COVID-19 vaccine recommendations. Regression discontinuity analyses showed that birth-year-based vaccination policy, including routine vaccination opportunities, was significantly associated with recommendations to others for measles and HPV vaccines, independently of perceived benefits and risks. Conclusion: To improve vaccination coverage from a public health perspective, it is important for physicians to provide effective vaccination recommendations to the general public on a broader scale; however, it is also necessary to address the heterogeneity in vaccine-specific recommendation patterns among physicians, as observed for COVID-19. Routine vaccination opportunities may increase vaccination coverage not only through the routine vaccination program itself but also through peer effects among the general public. Vaccination policy may therefore influence vaccination coverage not only in the current generation but also in future generations. Designing vaccination policy should consider its long-term impact on future vaccination coverage as well as herd immunity.

Background: ChAdOx1 nCoV-19 remains a cornerstone COVID-19 vaccine in sub-Saharan Africa, yet population-specific molecular responses are understudied. We examined peripheral blood ACE2 and TMPRSS2 expression, total RNA concentration, and coagulation indices in Nigerians >=6 months post-vaccination. Methods: In a case-control study in Port Harcourt, Nigeria, 51 ChAdOx1-vaccinated adults and 51 age/sex-matched unvaccinated controls provided venous blood for RNA extraction, qRT-PCR, and coagulation assays. Multivariable linear models assessed effects of vaccination, sex, and age on molecular parameters. Results: Vaccinated participants had 37% lower total RNA concentration than controls (4.02 +/- 0.09 vs 6.38 +/- 0.14 ng/uL, p<0.0001). ACE2 and TMPRSS2 expression did not differ by vaccination status overall. However, TMPRSS2 showed a significant sex-by-treatment interaction (p=0.011): vaccinated females had higher expression than vaccinated males. GAPDH expression varied by vaccination status and showed a three-way interaction with sex and age (p=0.027). Coagulation indices were unchanged. Conclusions: At >=6 months post-ChAdOx1, Nigerians show reduced peripheral blood RNA without sustained ACE2/TMPRSS2 upregulation. The sex-specific TMPRSS2 pattern suggests hormone and vaccine interactions previously unreported in African cohorts and highlights the need for sex-disaggregated molecular surveillance. Region-specific reference gene validation is recommended for Nigerian transcriptomic studies.

Background: While haematological and coagulation changes following AstraZeneca vaccination have been described, the molecular mechanisms linking TMPRSS2 expression to coagulation remain underexplored, particularly in African populations. Methods: In this case-control study, 102 adults (51 vaccinated with AstraZeneca >=6 months prior, 51 unvaccinated controls) aged 18-65 years in Port Harcourt, Nigeria, were evaluated. Full blood count (Sysmex XN-1000), PT/aPTT (Erba Mannheim), RNA concentration, and qRT-PCR for ACE2/TMPRSS2 (normalized to GAPDH) were performed. Pearson correlations and t-tests were conducted (SPSS v26, p<0.05). Results: Because primary between-group comparisons from this cohort have been reported previously, the present analysis focused on correlation patterns. A statistically significant inverse correlation was observed between TMPRSS2 Ct values and activated partial thromboplastin time (aPTT) among vaccinated participants (r = -0.325, p = 0.0202), whereas no corresponding association was detected in unvaccinated controls. Sex-specific differences in TMPRSS2 expression were also observed. No participant reported severe thrombotic or haemorrhagic complications at the time of recruitment. Conclusion: This secondary analysis identified a statistically significant inverse correlation between TMPRSS2 Ct values and aPTT among AstraZeneca-vaccinated individuals. Although causality cannot be inferred, the findings suggest a potential relationship between TMPRSS2 expression and haemostatic pathways in the post-vaccination setting. Larger longitudinal studies are required to validate the observation and clarify its biological significance.

Purpose: Human papillomavirus (HPV) infection and HPV-associated diseases impact health-related quality of life (HRQOL). We aimed to measure HRQOL in women with HPV-positive test results, cervical intraepithelial neoplasia (CIN), or cervical cancer in Latin America. Methods: We enrolled women aged 18 to 75 years from Bolivia, Colombia, Honduras, Peru, and Uruguay. We used the EQ-5D-5L instrument to calculate EQ-5D index scores with country-specific sets of health preferences when available. We calculated medians and interquartile ranges (IQR) of EQ-5D index and conducted an exploratory analysis comparing HRQOL loss by diagnosis and country with a gamma regression. Results: We present results from 1,073 participants. Median age was 43 years (IQR: 34-52). The EQ-5D index scores by diagnosis were as follows: HPV-positive test alone 0.906 (standard deviation [SD]: 0.129), CIN1: 0.891 (SD: 0.136), CIN2: 0.892 (SE: 0.124), CIN3: 0.870 (SD: 0.138), and cervical cancer: 0.737 (SD: 0.268). HRQOL was associated with age, diagnosis, and country; there was a significant decreasing trend in HRQOL with worsening health state. Women with cervical cancer had a 3.19-fold higher HRQOL loss than women with an HPV-positive test alone or CIN1. We also found significant differences in HRQOL loss across countries, after adjustment for diagnosis and sociodemographic factors. Conclusion: HRQOL decreased with diagnosis severity, with significant differences between countries. To conduct cost-effectiveness modeling on HPV preventive interventions, obtaining accurate HRQOL estimates is essential. This process should consider the diverse local health preferences influenced by sociodemographic and cultural factors, as well as the population's beliefs and experiences regarding health.

Background In November 2023, the Joint Committee on Vaccination and Immunisation advised the UK government that a targeted, opportunistic vaccination programme using 4CMenB to prevent gonorrhoea primarily in gay, bisexual and other men who have sex with men (GBMSM) at higher risk of infection should be introduced in sexual health services (SHSs). Data on the acceptability of 4CMenB vaccination and factors influencing uptake were needed. Methods Three focus group discussions (FGDs) were conducted with 17 GBMSM aged [&ge;]18 years, resident in England, who self-reported bacterial sexually transmitted infection or [&ge;]5 sexual partners in the previous 12 months. One FGD with five sexual healthcare professionals (HCPs) was conducted. Data were analysed using reflexive thematic analysis. Themes were organised using the Vaccine Uptake Continuum and interpreted using the Social Ecological Model. Results Acceptability of 4CMenB vaccination was high among GBMSM and HCP participants. GBMSM described vaccination as supporting sexual wellbeing and reducing anxiety about gonorrhoea, particularly when positioned alongside existing prevention strategies like HIV pre-exposure prophylaxis and Doxycycline post-exposure prophylaxis. While the estimated effectiveness of ~30-35% was perceived as modest, it did not deter acceptability, but reduced willingness to actively seek vaccination. Structural constraints, including limited SHS capacity, appointment availability, and restrictive eligibility criteria, were identified as barriers to equitable uptake. Community-based and outreach delivery models were widely supported as strategies to improve access. HCPs drew on experience from mpox vaccination to anticipate implementation challenges, emphasising the need for clear guidance, staff training, and sustainable resourcing. Mixed views were expressed regarding additional protection against meningitis, which was generally considered a secondary influence on decision-making. Conclusions 4CMenB vaccination for gonorrhoea was acceptable for both GBMSM and HCPs; however, uptake is likely to depend on ease of access, clear communication, and system-level support. Addressing structural constraints and supporting community-based delivery may help achieve equitable delivery of 4CMenB.

Cross-reactive antibodies against dengue virus are known to cause antibody-dependent enhancement (ADE) of infection or disease severity under specific conditions. In our previous study, we showed that primary immunization with the COVID-19 vaccine induces induces cross-reactive IgG causing ADE against dengue. In the present study, we investigated the influence of IgG Fc-glycosylation (analyzed by LC-MS/MS) on ADE mediated by cross-reactive IgG against dengue from IgG against SARS-CoV-2. We found a clear correlation between anti-DENV2 E IgG2 galactosylation and the ADE capacity of cross-reactive IgG against dengue in individuals vaccinated against COVID-19. IgG2 sialylation increased over time; however, it was not correlated with ADE capacity. This phenomenon was restricted to IgG2, whereas anti-DENV2 E IgG1 Fc-glycosylation remained stable after COVID-19 vaccination.

Colombia has been internationally recognised as a paradigmatic case of vaccine confidence crisis since the 2014 Carmen de Bolivar event, and national HPV vaccination coverage remains far below the World Health Organization 2030 target. Most published evidence focuses on female adolescents and on cervical cancer; the perception of the HPV vaccine in university-age populations of both sexes--and across the broader spectrum of HPV-attributable disease--remains comparatively understudied. We aimed to describe the influence of biopsychosocial determinants on HPV vaccine perception among university students of both sexes in Cucuta, Norte de Santander, Colombia. We conducted a cross-sectional study with a mixed quantitative-qualitative approach in 2024 among four universities (Universidad de Santander, Universidad Francisco de Paula Santander, Universidad de Pamplona and Universidad Libre; combined enrolment 21,033 students). Using convenience sampling stratified by institution, 750 actively enrolled undergraduate students of both sexes (18-60 years) completed a structured online questionnaire adapted from previously validated instruments. The instrument captured sociodemographic information, HPV knowledge and HPV vaccine perception. Data were analysed using Students t-test, one-way analysis of variance, Tukey post-hoc tests, effect sizes and 95% confidence intervals, with a 0.05 significance threshold. Of 750 respondents, 54.2% were women, 61.3% were under 20 years of age, and 75.1% attended public universities. HPV knowledge was high in 39.2%, intermediate in 42.4% and low in 18.4%; women and students aged 26 years or older displayed higher knowledge. Although 91.2% had heard of HPV and 82.5% knew that both sexes could acquire it, recognition of clinical manifestations and complications was uneven: cervical cancer 51.7%, penile cancer 30.5%, vaginal warts 45.9% and warts in the penis, larynx, anus or rectum 34.0%. Vaccine-specific knowledge was low in 77.1%, with men disproportionately represented (85.9% versus 69.5% in women). Overall positive perception of HPV vaccination was 66.6%, slightly higher in women (68.8%) than men (63.9%), in students aged 26 years or older (70.1%) and in students from private universities (68.1% versus 65.9%). Inferential analysis identified sex (Cohens d = -0.357), type of university (d = 0.189) and HPV knowledge (partial eta-squared = 0.096) as the only significant determinants. Age, socioeconomic stratum, age at sexual debut and vaccine-specific knowledge did not reach meaningful significance. HPV vaccine perception was predominantly positive but conditioned by three biopsychosocial determinants, with HPV knowledge as the primary driver. The persistent gender gap reflects historical anchoring of HPV messaging in cervical disease and female-targeted campaigns. Public-health strategies should adopt comprehensive, gender-inclusive educational interventions that explicitly visibilise non-cervical HPV-related cancers and address both sexes from a common evidence base.

The Africa CDC's New Public Health Order aims to improve health, regional self-reliance, and health security by boosting local vaccine manufacturing capacity from <1% in 2022 to 60% by 2040. Although multiple studies have examined vaccine manufacturing capacity in Africa, none quantified future vaccine manufacturing capacity requirements and complexities in capacity planning. We fill this gap by integrating a deterministic approach and discrete event simulation (DES) to estimate vaccine manufacturing capacity requirements and assess the impact of process variability and uncertainty on manufacturing capacity needs. A total of 51 experts were interviewed to design vaccine-specific supply chain networks, collect, and validate data on process-specific parameters. Our findings reveal that deterministic approach provides optimistic capacity estimates but realistic estimates of capacity requirements is possible using DES as it captures variability and uncertainty inherent in vaccine manufacturing. Further evidence shows that reduction in batch yield at any manufacturing stage significantly reduces throughput, while the impact of high batch deviations is vaccine specific and dependent on the manufacturing stage. For multi-antigen vaccines, capacity estimation is complicated by the amplification of process variability, and synchronization of batch arrival and volume. Finally, we estimated that 11 vaccine manufacturing facilities staffed by approximately 8,599 fulltime employees, of whom 43% are technical personnel, would be required to achieve Africa's 2040 ambition. Overall, adopting a system-level perspective, maintaining a high, reliable, and sustainable manufacturing process yield and quality, are important when establishing new vaccine manufacturing facilities.

Background: The N-terminal ectodomain (NTE) of the SARS-CoV-2 membrane (M) glycoprotein is a short, flexible region that remains exposed on the virion surface and exhibits immunogenic potential across multiple coronaviruses. Despite its small size and conformational plasticity, this region contains conserved linear epitopes that may serve as practical surrogates for full-length proteins in serological diagnostics. Objective: To develop and evaluate a synthetic peptide-based diagnostic assay targeting the NTE of the SARS-CoV-2 M protein. Methods: Epitope prediction, peptide synthesis, and antibody affinity assays were performed to design homomultivalent peptide analogs that exploit avidity effects through disulfide polymerization. The resulting peptide antigens were tested in an enzyme-linked immunosorbent assay (ELISA) using clinical samples from RT-PCR-confirmed COVID-19 patients and biobanked controls. Results: The selected peptide analogs (M1, M1i, M1s) corresponded to a conserved surface-exposed motif of the SARS-CoV-2 M protein. Polymeric M1 exhibited a twofold gain in apparent affinity (Kdapp = 4.33 nM) compared with the monomeric form (Kdapp = 8.00 nM). Clinical validation using 1,222 patient samples yielded a sensitivity of 95.26% and specificity of 52.27%, with an overall diagnostic accuracy of 88.70%. Conclusion: The M peptide analogs demonstrate that synthetic peptide antigens can serve as stable, high-sensitivity surrogates for whole-protein assays. This design principle may be applied to other emerging pathogens where rapid assay development and scalability are critical. Keywords: Peptides, Antibodies, COVID-19, Enzyme-Linked Immunosorbent Assay, Protein Binding

Objectives: Several novel tuberculosis vaccines (NTVs) are being evaluated in clinical trials. Understanding the perception of acceptability and confidence in NTVs will inform implementation strategies. Methods: We conducted a cross-sectional survey to assess acceptability in and confidence of NTVs among adult persons with HIV (PWH) at two clinics in KwaZulu-Natal province, South Africa. We evaluated associations between acceptability and sociodemographic factors. Results: Among 225 PWH, 112 (50%) responded "Definitely yes, as soon as available", 86 (38%) "Definitely yes, but wait [&ge;]6 months to receive it" and 27 (12%) "Unsure, leaning yes, any timeline" to an NTV. Acceptability was associated with perception of tuberculosis's (TB's) importance in the community, perceived risk of contracting TB, personal history of TB, not currently living with someone with TB, incompatibility of vaccines with religion, and unemployment. Most participants were confident in the vaccine's potential safety (110, 93%) and effectiveness (221, 98%). Participants preferred to receive information from government, community, and health entities, or internet, and vaccination at community settings over public health facilities. Conclusions: NTV acceptability was high amongst these PWH, and they preferred community-based delivery models. Our findings may inform strategies to increase implementation of NTVs among PWH in South Africa.

Abstract Background: Human papillomavirus (HPV) infection is a leading cause of cervical cancer globally, disproportionately affecting women in developing countries like Ghana. Despite the recent introduction of national HPV vaccination programs, vaccine uptake among young adults remains suboptimal. This study aimed to assess the levels of knowledge, attitudes, and perceived barriers toward HPV, cervical cancer, and vaccination among undergraduate students in Ghana to inform future public health interventions. Methods: A cross-sectional study was conducted among 699 undergraduate students at the University of Cape Coast, Ghana. A multistage stratified random sampling technique was employed to ensure disciplinary representation. Data were collected using a validated, semi-structured digital questionnaire covering socio-demographics, knowledge of HPV, attitudes toward vaccination, and perceived barriers. Descriptive statistics were utilized to summarize findings. Chi-square tests were performed to assess bivariate associations, and binary logistic regression analysis was conducted to identify predictors of good knowledge and positive attitudes toward vaccination, with statistical significance set at p < 0.05. Results: 51.9% of students demonstrated good knowledge of HPV and vaccination. A significant gender disparity was observed: while male students displayed higher levels of clinical knowledge, female students held significantly more positive attitudes toward vaccination (p < 0.05). Major barriers included profound social stigma, with 77.9% of students expressing concern over partner perception and 65.6% reporting embarrassment regarding the association between the vaccine and sexually transmitted infections. Misconceptions were prevalent, with 46.6% of participants incorrectly believing the vaccine could cure existing infections. Conclusion: A clear knowledge-attitude gap exists among Ghanaian undergraduates, complicated by pervasive psychosocial barriers. Current vaccine delivery models, which often center on reproductive health or STI clinics, inadvertently reinforce stigma. To improve vaccination coverage, public health initiatives must transition toward a stigma-neutral model of care that integrates HPV immunization into routine primary health services, framing it as a preventive cancer-fighting strategy rather than a sexual health intervention.

The diversity within H1 and H3 subtype influenza A viruses (IAV) in swine prevents effective vaccine control approaches with inactivated whole-virus vaccines. We addressed the challenge of controlling co-circulating hemagglutinin (HA) clades of swine IAV with the development of a multivalent mRNA-lipid nanoparticle (LNP) vaccine expressing 8 HA proteins to maximize genetic coverage. We applied a computational approach to select eight HA genes that represented 95% of the observed IAV detected in the United States between 2022 and 2025. Piglets were vaccinated and boosted intramuscularly with either individual HA mRNA-LNP or an 8-HA multivalent mRNA-LNP. Serum was collected to evaluate systemic antibody levels. Twenty-one days post-boost, pigs were challenged with a field relevant H1 1A.3.3.3-c3 IAV strain. The 8-HA multivalent mRNA-LNP vaccine induced neutralizing antibodies against all eight antigens and vaccinees were protected against lung lesions, with lesion scores similar to non-challenged animals. Homologous monovalent vaccination significantly reduced IAV detection in nasal secretions and in the lungs. Heterologous monovalent vaccination was not cross-protective but did not induce vaccine-associated enhanced respiratory disease. We provide evidence that monovalent and multivalent mRNA-LNP influenza vaccines elicited neutralizing antibody responses in pigs and protected against viral challenge. The versatility and capacity for rapidly updating the mRNA-LNP vaccine platform make it an appealing tool to improve animal health and minimize the circulation and diversity of IAV in swine. ImportanceInfluenza A virus is an important respiratory pathogen in swine, and zoonotic transmission of swine strains to humans remains a public health risk. Control strategies against IAV in swine herds rely heavily on biosecurity measures and vaccination. However, the antigenic diversity of IAV circulating in swine challenges current vaccination programs, and there is a need for broadly protective vaccines or platforms that can rapidly update components to reflect circulating diversity. mRNA-LNP vaccines have emerged as promising vaccine platforms, offering simultaneous delivery of multiple antigens, rapid development, scalable manufacturing, and potent immunogenicity. In this study, we assessed the immunogenicity and protective capacity of monovalent and multivalent mRNA-LNP vaccines encoding eight representative IAV HA antigens. To our knowledge, this is the first study to objectively select multiple representative endemic swine IAV strains by quantifying genetic diversity within the phylogeny and to apply this selection to rationally design and evaluate a multivalent HA mRNA-based influenza vaccine in the swine model.

Introduction Respiratory syncytial virus (RSV) is an important cause of respiratory disease in older adults and adults with chronic medical conditions, contributing substantially to the healthcare burden in Italy. The availability of effective RSV vaccines provides an opportunity to reduce RSV-related morbidity, mortality, and healthcare costs in populations at high risk of severe disease. This study evaluates the potential public health impact and cost-effectiveness of vaccination using mRNA-1345 administered as a single dose compared with no vaccination in Italian high-risk adults aged 60-74 years and all adults aged [&ge;]75 years. Methods A static decision-analytic model was developed to project clinical and economic outcomes over a 5-year time horizon. Economic outcomes were evaluated from the Italian National Health Service (Servizio Sanitario Nazionale, SSN) perspective. Model inputs were informed by the most recent Italian epidemiological, clinical, and economic evidence, supplemented by published international data when necessary. Deterministic, probabilistic, and scenario analyses were conducted to assess the impact of uncertainty in model inputs and assumptions on the study results. Results Vaccination with mRNA-1345 in high-risk adults aged 60-74 years and all adults aged [&ge;]75 years was projected to avert over 19,800 hospitalizations, 4,000 emergency department visits, 381,000 outpatient visits, 6,000 RSV-attributable deaths, and 212,000 antibiotic prescriptions compared with no vaccination over a 5-year period. The total incremental cost of {euro}1,143 million and the additional 47,477 QALYs gained resulted in an ICER of {euro}24,078, which was below the commonly referenced willingness to-pay range of {euro}33,000-40,000 per QALY gained. Sensitivity analyses confirmed robustness of the analysis results. Conclusions Vaccination with mRNA-1345 is a cost-effective strategy for the prevention of RSV in high-risk adults aged 60-74 years and all adults [&ge;]75 years in Italy and has the potential to provide substantial public health benefits.

Introduction: Synthetic oligopeptides provide a rapid and cost-efficient approach to developing antibodies and diagnostics for emerging viral variants. Methods: This study computationally and experimentally characterized a synthetic peptide analog of the SARS-CoV-2 spike subdomain 2 major disulfide loop (SD2MDL), designated S621 (CPVAIHADQLTPTWRVYSTC). Binding affinity was computationally estimated using the Heuristic Affinity Prediction Tool for Immune Complexes (HAPTIC), while experimental validation was performed using enzyme-linked immunosorbent assay (ELISA) with rabbit-derived antipeptide antibodies. Clinical diagnostic accuracy testing was done using plasma samples from RT-PCR-confirmed COVID-19 patients and pre-COVID-19 controls. Results: S621 demonstrated nanomolar binding affinity (Kdapp = 1.14 nM) and high avidity (3.67 nM), closely matching HAPTIC predictions (3.54 nM). Diagnostic evaluation yielded a sensitivity of 89.92% and specificity of 27.79%, corresponding to an overall accuracy of 71.79%. Discussion: These findings demonstrate that a single synthetic peptide derived from a conserved spike subdomain can function as a high-affinity surrogate for full-length antigens, supporting its potential application in rapid peptide-based immunodiagnostics.

Background: Responses to vaccination programmes vary widely, with contrasting perceptions of benefits and harms. This study aimed to provide a contemporary characterization of distinct groups of respondents by moving past traditional "pro" versus "anti" vaccination binaries. The results captured the complex spectrum of vaccine sentiment to directly inform the next strategic phases of a multi-year public health campaign regarding vaccine perceptions and behaviours within the diverse population of London. Methods: Data came from the 2024 Why We Get Vaccinated survey conducted in London, UK using a community-based participatory approach. Participants answered vaccine-related questions based on previous vaccine uptake, future willingness to vaccinate (WTV), perceived effectiveness, and safety concerns. We employed Bayesian Binomial logistic regression to identify sociodemographic predictors of individual responses. Subsequently, a Bayesian Nonparametric Latent Class Analysis (BNP-LCA) using a Dirichlet Process Mixture model was used to identify distinct groups and their sociodemographic compositions. Results: The LCA identified five distinct classes. Class 1, the Consistent Uptakers (37.2%), exhibited high vaccine uptake (89.2%) and near-universal belief in efficacy (98.0%) with minimal safety concerns. Class 2, the Concerned Uptakers (17.5%), maintained high WTV (72.2%) despite significant concerns regarding adverse effects (71.2%). Class 3, the Consistent Refusers (18.4%), demonstrated uniform rejection of vaccines and high levels of safety concern (74.4%). Class 4, the Unconcerned Refusers (20.8%), acknowledged vaccine effectiveness (98.4%) but showed very low uptake (1.7%), possibly due to low perceived personal risk rather than active resistance. Class 5, the Undecided/Uninformed (6.0%), was characterized by pervasive uncertainty and "Don't Know" responses regarding efficacy (80.3%) and vaccination intent. Sociodemographic analysis revealed that residential instability, i.e renting, and caregiving responsibilities were significant barriers to uptake, while older age and retirement were the strongest positive predictors. Conclusions: Vaccine behaviour and attitudes in London are a multidimensional construct. The discovery of "Unconcerned Refusers" and "Concerned Uptakers" highlights that "one-size-fits-all" public health campaigns may not be effective. Tailored public health messaging is required to address the specific uncertainties and socio-economic barriers identified across these distinct profiles. These findings provide a functional framework that will directly guide upcoming resource allocation, messaging adaptations, and policy decisions for the next iterations of the ongoing campaign

Introduction Hepatitis E virus (HEV) in pregnancy is associated with high maternal and perinatal morbidity and mortality. The safety and efficacy of the recombinant protein HEV vaccine (HEV239, Hecolin) have been established in non-pregnant adult populations but there is limited information among pregnant women. This trial has two co-primary objectives: 1) to assess pregnancy-related and/or serious safety events among pregnant women between 14 and 34 weeks of gestation receiving two Hecolin doses four weeks apart compared to placebo recipients, and 2) to determine immune non-inferiority of pregnant recipients of two Hecolin doses four weeks apart compared to non-pregnant women. Methods and Analysis This is a multi-site, randomized, observer-blinded, placebo-controlled vaccine safety and immunogenicity trial in pregnant women and non-pregnant women of reproductive age in Karachi, Pakistan. A total of 2,358 healthy women will be enrolled, including 2,208 pregnant women between 14 and 34 weeks of gestation, who will be randomized in a 1:1 ratio (stratified by gestational age, 14-27 and 28-34 weeks) to receive either Hecolin or a normal saline placebo in two doses administered 1 month apart during pregnancy and a third dose administered postpartum, approximately 5 months after the second dose. A third arm of 150 non-pregnant women aged 16-45 years will receive Hecolin on 0, 1, and 6 months. The co-primary outcomes will be (i) the proportion of pregnancy-related AESIs and SAEs in pregnant participants from the first dose until the end of study follow-up, compared with placebo, and (ii) the geometric mean concentration (GMC) of anti-HEV IgG at four weeks after the second dose, comparing pregnant vaccine recipients with non-pregnant vaccine recipients (non-inferiority margin of 0.67 for the GMC ratio). Immunogenicity will be evaluated in a pre-specified subset of 300 participants receiving Hecolin, including 150 pregnant participants and 150 non-pregnant participants. Secondary outcomes will include maternal, neonatal, and infant safety outcomes, as well as immunogenicity according to the number of Hecolin doses received and the trimester of vaccination. Ethics and Dissemination The trial was approved by the National Bioethics Committee (NBC) of Pakistan (Reference number: 4-87/NBC-910), the institutional Ethics Review Committee (ERC) of the Aga Khan University (Reference number: 8298), and the Institutional Review Board (IRB) of the International Vaccine Institute (IVI) (Reference number: 2022-007). All participants will provide written informed consent in accordance with Good Clinical Practice. The results will be submitted to World Health Organization (WHO) Strategic Advisory Group of Experts in Immunization (SAGE), and disseminated through conference presentations, and peer-reviewed publications.

1.Virus-like particles (VLPs) formed from the murine polyomavirus major capsid protein VP1 are widely used as vaccine antigens and are being explored as nucleic acid and drug delivery vehicles. However, the factors controlling distinct VP1 capsid morphologies remain unclear. We investigated in vitro VP1 assembly with tRNA across NaCl concentrations of 0.15-1.0 M and tRNA mass ratios of 1:1-1:80 (w/w) using SEC-HPLC, transmission electron microscopy, and dynamic light scattering. Two competing assembly pathways were identified. At low ionic strength ([&le;]0.15 M NaCl), nucleic acid-templated assembly produced compact, tRNA-filled T=1 VLPs ([~]28-30 nm). Assembly was maximal at tRNA ratios of 1:10-1:20, whereas excess or insufficient tRNA reduced yields. Increasing NaCl to 0.30 M lowered T=1 yields by 68-97%, and no T=1 particles were detected at [&ge;]0.5 M NaCl. Conversely, high ionic strength ([&ge;]0.5 M NaCl) promoted template-independent formation of hollow T=7 VLPs ([~]55-60 nm). T=7 assembly was inhibited by tRNA and was highest without nucleic acid. At 1.0 M NaCl, reducing the tRNA ratio from 1:40 to 1:80 increased T=7 yield more than 11-fold, while removing tRNA produced the greatest assembly efficiency. Kinetic analyses further showed that VP1 concentration and ionic strength regulate nucleation and assembly rate in the template-driven pathway. These findings show that electrostatic interactions govern pathway selection between tRNA-templated T=1 assembly and salt-driven T=7 self-assembly, providing practical guidance for controlling capsid morphology and cargo loading in VLP-based applications.

We have used synthetic biology recombination methods in yeast to build herpes simplex virus type-1 (HSV-1) and human cytomegalovirus (HCMV) genomes from multiple fragments. The genomes were built using transformation-associated recombination (TAR) in yeast, by virtue of overlapping sequences between the different fragments. This study demonstrates the successful assembly of the Epstein-Barr virus (EBV) genome. We used as the model genome, the Akata Burkitts lymphoma genome, specifically the BX1 genome which encodes a neomycin selectable marker and a GFP expression cassette in the BXLF1 region. The 171.3 kb genome was first deconstructed into 11 fragments in silico, each having 80 bp overlapping sequence between the fragments. The 11 fragments (TAR 1 to TAR 11) were cloned using TAR in yeast, analyzed by restriction enzyme analyses and Nanopore sequencing to validate the cloned fragment. The EBV genome was built in two stages: TAR fragments 1 to 6 and TAR fragments 7 to 11 were assembled to generate two half-genomes. The whole genome (TAR 1-11) was then assembled by joining TAR 1-6 with TAR 7-11. Complete EBV genomes were examined by PCR assays and restriction enzyme analyses and then transfected into HEK-293 cells to generate virus producer cell lines. The HEK-293 cell clones were tested for virus production following lytic induction using baculovirus transduction of Zta, Rta and glycoprotein B (BALF4). The supernatants from these induced cells were harvested and used to infect Raji cells. This analysis revealed a significant number of cells displaying strong GFP fluorescence indicative of infectious virus. We used this supernatant virus to infect primary B cells and were able to derive lymphoblastoid cell lines (LCL) indicative of the ability of this virus to transform B cells. We tested this method for engineering different mutations. Two mutations were made, one in Zta and the other in the small capsid protein (BFRF3). Mutations were engineered in the TAR plasmid in which the genes reside and after sequence validation, assembled into the TAR 1-6 half genome and then the TAR 1-11 genome, which was used to generate HEK-293 cell clones. For the {Delta}Zta cell lines, we could detect virus in the supernatants only if baculovirus expressing Zta in trans was included, this {Delta}Zta EBV virus could transform B cells. The small capsid protein (BFRF3) decorates the capsid shell and is required for capsid assembly in a self-assembly system. When the HEK-293 cell clones were induced using co-expression of Zta, Rta and gB, no virus was detected in the culture supernatants. However, if we provided BFRF3 in trans using baculovirus expressing this protein, virus was detected in the supernatants. This provides the first report of the essential role of the small capsid protein in EBV-infected cells.

Background Human papillomavirus (HPV) vaccination is an effective cancer prevention strategy, yet HPV vaccine awareness remains uneven across sociodemographic groups. In the current digital information environment, awareness may be shaped not only by access to health information but also by exposure to false or misleading health information, difficulty evaluating information accuracy, and echo-chamber dynamics on social media. Objective This study examined associations between perceived exposure to false or misleading health information on social media, difficulty determining whether social media health information is true or false, perceived echo-chamber exposure, and HPV vaccine awareness among U.S. adults. Methods We analyzed nationally representative Health Information National Trends Survey data using survey-weighted descriptive statistics and logistic regression models. The analytic sample included 2,371 respondents, representing a weighted population of 49.2 million U.S. adults. The outcome was HPV vaccine awareness. Primary predictors included perceived exposure to false or misleading health information on social media, difficulty determining whether social media health information was true or false, and perceived same-view health network exposure on social media. Models adjusted for age, sex, race/ethnicity, education, household income, rurality, and Census division. Results Overall, 60.37% of respondents reported HPV vaccine awareness. Most respondents reported encountering false or misleading health information on social media, with 45.57% reporting "some" and 32.83% reporting "a lot." In unadjusted models, greater perceived exposure to false or misleading health information was associated with higher odds of HPV vaccine awareness. After adjustment, respondents reporting "some" false or misleading health information had significantly higher odds of HPV vaccine awareness compared with those reporting none (AOR=2.40, 95% CI: 1.06-5.43), while the association for "a lot" was marginal (AOR=2.29, 95% CI: 0.97-5.38). Difficulty identifying true versus false social media health information and perceived echo-chamber exposure were associated with HPV vaccine awareness in unadjusted models but were attenuated after adjustment. HPV vaccine awareness was substantially higher among females and respondents with higher educational attainment, and lower among Hispanic, non-Hispanic Asian, and non-Hispanic other respondents compared with non-Hispanic White respondents. Conclusions HPV vaccine awareness is associated with both digital health information exposure and persistent sociodemographic inequities. Greater perceived exposure to misleading health information may reflect broader engagement with health-related content on social media, where accurate and inaccurate information coexist. Public health communication strategies should address misinformation vulnerability while expanding accurate, culturally responsive HPV vaccine messaging across digital platforms.

Cervical cancer remains a major public health challenge in Indonesia, with Human Papillomavirus (HPV) infection responsible for nearly all cases. The government has integrated HPV vaccination into the national School Children Immunization Month (BIAS). However, out-of-school (OOS) girls remain difficult to reach because they are not covered by the school-based vaccination platform. Micro-costing approach was used to estimate both economic and financial costs of delivering HPV vaccination to OOS girls in Bekasi City, Bandar Lampung City, and Bangka District. This study aimed to estimate the cost of delivering a single-dose HPV vaccination program to OOS girls and to project the national-level cost of scaling up the program. Costs were categorized into outreach efforts, such as community mobilization and identification of girls, and delivery components including logistics, storage, and administration. A scale-up costing analysis was also conducted to estimate national-level costs. The results show that the cost per vaccinated OOS girl aged 11 ranged from US$ 22.77 to 38.55, while the financial cost range from US$ 12.70 to 29.50, varied by implementation context, with outreach activities, transportation, and vaccine delivery supplies identified as the main cost drivers. The estimated national annual economic and financial cost of scaling-up HPV vaccination for OOS girls are US$ 644,215.23 and US$ 483,138.35, respectively. Our national cost estimates show that reaching and vaccinating OOS girls accounts for less than 3% of the national immunization budget. Factors such as local context and implementation challenges should be recognized as they may directly influence the cost and hinder the program scale-up.