Search Results (1,078)

Live-attenuated vaccines face a critical challenge in balancing immunogenicity with safety. To address this, we engineered programmable finite-replicated organisms (FROs) by depositing a limited number of indispensable components (such as noncanonical amino acids, ncAAs) within the cell, consuming the coenabling precise control of bacterial replication capability while preserving antigenic breadth. Two strategies were adopted to achieve the following purposes: (1) encoding ncAA in essential genes; (2) encoding ncAA in antitoxin of toxin–antitoxin (TA) systems. As noncanonical amino acids, 3,5-dichlorotyrosine (Cl2Y) was encoded by the amber codon (TAG) and inserted into the essential genes (e.g., serS, murG, and dnaA) or antitoxin genes. After optimizing expression and the number of amber codons in the storage genes, the FRO cells can grow up to six generations, achieving amplification approaching 100 times after depletion of the ncAA in the growth medium. The escape frequencies are 10⁻⁵ to 10⁻⁷, which need to be optimized by combining multiple storage genes in the same genome in the future. This work holds the potential to amplify the amounts of antigens for vaccines, potentially accelerating the development of next-generation vaccines against antibiotic-resistant threats. Full article

Background: Sirolimus is an effective treatment for kaposiform hemangioendothelioma (KHE), a rare vascular tumor in children. However, its immunosuppressive properties raise concerns regarding the safety and efficacy of vaccinations during treatment. This study aims to evaluate the safety and immunogenicity of inactivated and live-attenuated vaccines administered to pediatric KHE patients undergoing sirolimus therapy. Methods: We conducted a prospective study involving 56 KHE children receiving sirolimus who were vaccinated during treatment. Data on vaccine-related adverse events were collected to assess safety. Immunogenicity was evaluated by measuring seroconversion or protective antibody titers against vaccines, including Hepatitis B, DTaP, and MMR. Results: Among 56 catch-up vaccinated children, no serious adverse events related to vaccination were observed. Mild local or systemic reactions occurred in a minority of patients. Serological analysis demonstrated that children with kaposiform hemangioendothelioma (KHE) receiving sirolimus therapy were able to generate and sustain robust protective antibody responses following vaccination. High seroconversion rates and antibody titers were observed for both inactivated vaccines (e.g., hepatitis B and DTaP) and live-attenuated vaccines (e.g., MMR). Protective antibody levels were maintained both within 3 months and beyond 6 months post-vaccination, indicating durable immunogenicity under sirolimus treatment. Conclusions: Vaccination during sirolimus therapy appears to be safe and immunogenic in children with KHE. These findings support the administration of both inactivated and live-attenuated vaccines under appropriate clinical monitoring in this rare patient population. Full article

High temperature typically decreases feed intake, milk production, and efficiency and increases metabolic disorders and health problems, greatly impacting farm economics. Supplements based on Saccharomyces cerevisiae have been suggested to benefit cows under heat stress, but effects on dairy cow performance are contradictory. This study aimed to evaluate the influence of heat stress on the effects of live yeast supplementation on the performance of dairy cows. Environmental temperature parameters were compared to two thermal humidity indices (THI1 and THI2) using wet bulb or dew point temperatures, as explanatory variables of dairy cow performance during the hot season. The experiment followed a randomized complete block design with 12 Holstein cows blocked by lactation number, days in milk, and milk production (two cows per block) and within each block, each cow was randomly assigned to a maize silage-based TMR with a concentrate mixture containing no yeast culture (Control) or 1 g/kg concentrate dry matter of a live yeast culture based on S. cerevisiae (Yeast) for 35 days. The experiment lasted for 35 d. Dry matter intake (DMI) was significantly higher for Yeast than it was for Control for all classes of temperature and THIs studied with an average increase of 2 kg DM per day, except for mean THI1 (from 54 to 60), for which the DMI was similar between treatments. Yeast promoted significantly higher milk yield than Control for all classes of daily maximum and mean temperature, averaging an increase of 4 kg of milk per day. Results suggest a more marked effect of temperature and indicate that yeast supplementation improved lactation performance of dairy cows exposed to hot weather. Full article

Influenza is a serious and global health issue, and it is a major cause of morbidity, fatality, and economic loss every year. Seasonal vaccines exist but are not very effective due to strain mismatches, delays in production, and antigenic drift. This comprehensive overview discusses the current situation of influenza vaccination, including the numerous types of vaccines—inactivated, live attenuated, and recombinant vaccines—and their effectiveness, efficacy, and associated challenges. It highlights the effects of the COVID-19 pandemic on the trends of influenza vaccination and the level to which innovation should be practiced. In the future universal influenza vaccines will be developed that target conserved viral antigens to provide long-term protection to people. In the meantime, novel vaccine delivery platforms, such as mRNA technology, virus-like particle (VLP), and nanoparticle-based systems, and less cumbersome and invasive administration routes, as well as immune responses are also under development to increase access and production capacity. Collectively, these innovations have the potential to not only reduce the global influenza epidemic but also to change the way influenza is prevented and prepare the world for a pandemic. Full article

The Peste des Petits Ruminants (PPR) live attenuated vaccines, the PPR virus (PPRV) Nigeria 75/1 strain (lineage II) and PPRV India Sungry 96 strain (lineage IV), currently used for control and eradication programme are very efficient vaccines as they provide the host, sheep and goats, a lifelong immunity after a single minimum recommended dose of 10^2.5 TCID₅₀/mL. Unfortunately, both live attenuated vaccines are thermolabile and their use requires maintaining the cold chain from the manufactory premises to the field as most PPR-infected regions are facing of hot climate, with poor infrastructure, and the maintenance of an effective cold chain remains a challenge. To address this challenge, efforts have focused on developing thermotolerant (ThT) PPR vaccines using different stabilisers and improving the freeze-drying process. This study aimed to define the criteria for the evaluation of the stability of ThT PPR vaccines. A total of 37 batches of freeze-dried PPR vaccines using the PPRV Nigeria 75/1 strain, including eight (8) and twenty-nine (29) vaccines labelled as ThT and conventional formulations, respectively, were tested to evaluate the stability at temperatures of 40 °C to simulate the field conditions in some hot climate regions. All the vaccine batches included in this study initially showed acceptable levels of residual moisture, below 3%, and titres above the minimum WOAH standard requirement of 10^2.5 TCID₅₀/mL. Following the incubation at 40 °C, 56.7% and 46% of the 37 vaccine batches tested retained titres above 10^2.5 TCID₅₀/mL on day 3 and day 5, respectively. These vaccines use stabilisers such as skimmed milk, lactalbumin–sucrose, trehalose and one unnamed product (which may be protected for patent). The mean of titre loss among the PPR vaccines maintaining titres above 10^2.5 TCID₅₀/mL was 0.78 log10 at day 3 and 0.99 log10 at day 5, suggesting a significant early degradation during the first 3 days. Based on these data, it is proposed that thermotolerant PPR vaccines should maintain a minimum titre of 10^2.5 TCID₅₀/mL for vaccine dose on day 5 post-incubation at 40 °C with a titre loss below 1 log10 per mL. Preliminary immunogenicity test results showed that the PPR ThT vaccine meeting this criterion could be used in the field without maintaining a cold chain for up to 3 weeks, offering a practical solution for vaccination in remote areas. Full article

Background/Objectives: This study aimed to develop a new attenuated live mumps vaccine strain and determine its biological properties and effectiveness. Methods: Plaque purification and amplification were performed in chicken embryo cells. Candidate live attenuated mumps MuV-365 strain sequencing was performed. After evaluating the potential neurotoxicity of the MuV-365 mumps strain, a preclinical safety evaluation of measles–mumps–rubella (MMR) live attenuated vaccine containing the MuV-365 strain was performed to support the registration and application of the MMR vaccine. Finally, mumps neutralization antibody titers and the concentration of anti-serum mumps-specific IgG were determined to evaluate the immunogenicity and efficacy of the MuV-365 strain and MMR vaccine in mice and rhesus monkeys. Results: The plaque of the PL-KUM main seed virus was screened, and strains whose sequences were highly homologous to RIT4385 (JL-5 derived) were selected to amplify. The candidate live attenuated mumps MuV-365 strain was then developed. Safety evaluation results indicated that the MuV-365 strain had no potential neurotoxicity, and the MMR vaccine containing the MuV-365 strain also showed no significant safety hazard. The immunogenicity of MuV-365 strain in BALB/c mice was not inferior to S79 and PL-KUM. After two doses of the MuV-365 strain, the concentration of anti-serum mumps-specific IgG of the MuV-365 strain was significantly higher than that of the S79 strain (p < 0.01). In rhesus monkeys, the MMR vaccine had good immunogenicity against measles and rubella after one dose, while immunogenicity against mumps improved after two doses. Conclusions: The developed MuV-365 strain was genetically stable, with adequate safety and immunogenicity. Full article

Human metapneumovirus (HMPV) is a major cause of acute respiratory tract infections, particularly in infants, young children, older adults, and immunocompromised individuals. Since its discovery in 2001, the virus has been recognized for its significant clinical and socioeconomic impact. Despite extensive research, no licensed vaccines or antiviral therapies are currently available for HMPV. This review aims to synthesize current knowledge on HMPV prevention and treatment, and to highlight promising avenues for future interventions. Several monoclonal antibodies (mAbs) targeting conserved epitopes of the HMPV fusion (F) protein have shown strong neutralizing activity in vitro and in animal models, although none have reached clinical trials. Vaccine development, including subunit, live attenuated, vector-based, and mRNA platforms, is progressing, with some candidates showing promise in adult populations. However, data in children, especially seronegative infants, remain limited. Antiviral research has explored repurposed drugs such as ribavirin and probenecid, along with novel agents like fusion inhibitors and T-cell-based immunotherapies, though none are yet approved. The development of safe, effective interventions—especially multivalent approaches targeting multiple respiratory viruses—remains a high priority. Continued research is essential to bridge the gap between preclinical promise and clinical application and to reduce the burden of HMPV infection worldwide. Full article

This study reviews the role of epidemiology in the United States in the late 19th and early 20th century, which led to recognition that poliomyelitis is an infectious disease and set the stage for subsequent developments in virology and immunology, the development of inactivated and live attenuated polio vaccines, and a dramatic worldwide decrease in poliomyelitis mortality and morbidity. Epidemiological studies in the United States were systematically reviewed from the mid-19th to early 20th centuries. Isolated cases and scattered small outbreaks of poliomyelitis in the mid-19th century led to epidemics of increasing size by the end of the century, causing public consternation, especially as the disease was considered “new” and had a predilection for young children. By the 1890s, the seasonal pattern of epidemics suggested that poliomyelitis might have an infectious etiology, but direct evidence of communicability or contagiousness was lacking, so an infectious etiology was not widely suspected until the early 20th century. Reports of bacterial isolations from spinal fluid and postmortem tissues suggested that poliomyelitis might be a bacterial disease, and simultaneous outbreaks of paralytic disease in humans and animals suggested a possible zoonotic basis. Although experimental studies showed that it was theoretically possible for flies to serve as vectors of poliovirus, and occasional cases of polio were likely caused by fly-borne transfer of poliovirus from human feces to human food, a fly abatement field trial showed convincingly that flies, whether biting or non-biting, could not explain the bulk of cases during polio epidemics. In conclusion, the early application of epidemiological evidence beginning in the late 19th century strongly suggested the infectious nature of the disease, distinct from previously identified conditions. Subsequent advances in virology and immunology from 1909 to 1954 proved that poliomyelitis was a viral disease with no natural animal host and made feasible the development of an inactivated trivalent poliovirus vaccine by Salk, and, subsequently, a live-attenuated trivalent poliovirus vaccine by Sabin. Full article

There is a need for safe and effective vaccines against the Venezuelan equine encephalitis virus that infects both humans and equines. However, development of a live-attenuated vaccine using the TC-83 strain has been hampered by substantial reactogenicity and the potential for neuroinvasion. In this study, we demonstrate that V4020, a new TC-83-based investigational VEEV vaccine with redundant safety features preventing neuroinvasion and reversion, exhibited no neuroinvasion potential in a murine model. Following subcutaneous or intramuscular administration, a subset of mice that received the TC-83 vaccine succumbed to central nervous system infection, with replicating virus detected in the CNS, demonstrating a low, yet detectable neuroinvasion potential of the TC-83 vaccine in vivo. Sequencing analysis of the TC-83 virus recovered from the brains identified a pseudoreversion of E2 R120I, as E2 R120 is known to confer attenuation for TC-83. In contrast, V4020 showed no evidence of virus in the CNS, highlighting one of the V4020 features, a new synonymous codon to minimize reversion to the wild-type residue. Overall, our study establishes V4020 as a rationally designed, safe vaccine candidate for VEEV with significantly reduced neuroinvasion risk. Full article

The varicella attenuated virus vaccine, developed in Japan in the 1970s, has dramatically reduced the number of pediatric chickenpox cases over the past 30 years due to its widespread use. However, a small number of cases of chickenpox, shingles, aseptic meningitis, and acute retinal necrosis caused by vaccine strains have been reported. There are also issues that need to be addressed, such as breakthrough infections and the persistence of the preventive effect of vaccination. In addition, there is the possibility of the emergence of revertants or mutations in the vaccine strain. In recent years, subunit vaccines have been developed, their immune-stimulating effects have been demonstrated, and they are being applied clinically. In addition, development of an mRNA varicella vaccine is underway. In this review, the history and impact of the varicella vaccine are overviewed, as well as its future challenges. Full article

Background: Abrupt dietary changes may disrupt gut microbiota populations and lead to gastrointestinal issues. This study aimed to determine the effects of live Bacillus pumilus SG154 or Lacticaseibacillus paracasei 327 postbiotic on fecal characteristics and microbiota populations of dogs following an abrupt diet change. Methods: Twelve healthy adult English pointer dogs (6.38 ± 2.75 yr) were used in a replicated 3 × 3 Latin square design to test the following treatments: (1) placebo (control; 250 mg maltodextrin/d); (2) live B. pumilus [5 × 10⁹ colony-forming units (CFU)/d]; and (3) L. paracasei postbiotic (100 mg; derived from 2 × 10⁹ CFU/d). Each period lasted 42 days, with the diet change occurring on day 28. Fecal samples were scored and analyzed for pH, dry matter content, and microbiota before and 2, 6, 10, and 14 days after the diet change. Results: The abrupt diet change increased (p < 0.01) fecal pH, increased (p < 0.01) the dysbiosis index, decreased (p < 0.0001) fecal dry matter, and led to a large shift in the fecal microbiota community. Fecal scores were lower (p < 0.05) in the B. pumilus group. B. pumilus reduced (p < 0.05) the relative abundance of fecal Prevotella and Muribaculaceae, while both treatments (B. pumilus; L. paracasei) increased (p < 0.05) the relative abundance of fecal Holdemanella. Conclusions: These results suggest that an abrupt diet change leads to large shifts in fecal microbiota and modified fecal characteristics. The supplementation with a B. pumilus probiotic and a L. paracasei postbiotic slightly altered the relative abundance of a few microbial taxa but was unable to attenuate most responses. Full article

Background/Objectives: Vaccine development for the prevention of ASF has been very challenging due to the extensive genetic and largely unknown antigenic diversity. Inactivated vaccines, using different inactivation methods and a variety of adjuvants, have been consistently inefficacious. Historically, animals recovering from an infection with an attenuated virus became protected from the development of a clinical disease caused by an antigenically related strain. Therefore, immunization of susceptible animals with attenuathe ted virus strains has become a common method of vaccination with the first two commercially available vaccines based on recombinant live-attenuated viruses (LAVs). An important limitation is that the efficacy of the LAV is restricted to those strains that are antigenically related and, in most cases, only provide protection against homologous strains. Due to the unknown antigenic heterogeneity among all ASFV field isolates, the development of broad-spectrum vaccines is a challenge. Besides the anecdotal data, there is not a large amount of information describing patterns of cross-protection between different ASFV strains. Methods: We evaluated the cross-protection induced by the ASFV live-attenuated vaccine ASFV-G-ΔI177L against different biotypes of ASFV and compared their genomic sequences to determine potential genetic mutations that could cause the lack of cross-protection. Results: Results presented here demonstrate different patterns of protection when ASFV-G-ΔI177L vaccinated pigs were challenged with six different ASFV field isolates belonging to different biotypes. Conclusions: The presence of cross-protection cannot be predicted solely by the classical methodology for genotyping-based B646L ORF only. Biotyping, considering the entire virus proteome, appears to be a more promising prediction tool, although additional gathering of experimental data will be necessary to fully validate it; until then, the presence of cross-protection needs to be confirmed in efficacy trials challenging vaccinated animals. Full article

Background: Brucellosis is a zoonotic bacterial disease primarily controlled through quarantine, culling, and vaccination. Live attenuated vaccines remain the most effective countermeasure, yet their application is limited by residual virulence and diagnostic interference. This study developed three rough-type attenuated Brucella melitensis mutants (G7, G8, G16) and evaluated their potential as DIVA (Differentiating Infected from Vaccinated Animals) vaccine candidates. Methods: Rough phenotypes were characterized through heat agglutination, acridine orange staining, and immunoblotting. Macrophage cytotoxicity was assessed via LDH release assays, while RT-qPCR analyzed macrophage activation capacity. Mouse infection and immunization-challenge experiments, complemented by histopathology, evaluated residual virulence and protective immunity. Antibody profiles were determined by ELISA, and DIVA capability was verified using LPS-coated ELISA. Results: G7 and G8 exhibited complete rough phenotypes, whereas G16 retained partial O-antigen (semi-rough). All rough mutants induced macrophage cytotoxicity and activation. The strains showed attenuated virulence with no viable bacteria recovered from spleens at 4 weeks post-inoculation. Histopathology revealed no liver lesions at 6 weeks post-inoculation. Immunized mice predominantly produced IgG2a-dominated Th1-type responses. The immune protection levels of G7 and G16 matched the reference vaccine M5–90Δ26, while G8 showed slightly lower efficacy. LPS-ELISA effectively differentiated vaccinated from infected animals via concurrent IgM/IgG detection. Conclusions: This study demonstrates that the rough-type B. melitensis mutants G7 and G16 serve as promising DIVA vaccine candidates, offering strong protection with low residual virulence while enabling serological differentiation between vaccinated and infected animals, highlighting their potential as effective vaccines for brucellosis control. Full article

Infectious bronchitis virus (IBV) remains a major threat to poultry health worldwide due to frequent genetic changes mainly driven by recombination and limited cross-protection between genotypes. In this study, we analyzed IBV strains collected from clinical outbreaks in Chile between 1986 and 2021 to assess the long-term impacts of live-attenuated vaccines (Massachusetts and 4/91) on viral evolution. Phylogenetic analysis of the S1 and N genes revealed four major lineages circulating in Chile—GI-1, GI-13, GI-16, and a novel monophyletic clade we propose as GI-31. The latter, identified in isolates from 1986 to 1988, is highly divergent (22–24%) from other known lineages, representing a previously unreported South American IBV variant. Despite widespread Mass vaccination, genetically distinct field strains circulated during the 1980s, facilitating potential recombination with GI-1 vaccine-derived strains, including evidence of shared ancestry with GI-11, an endemic lineage from Brazil. Non-recombinant GI-16, likely introduced from Asia, was detected in isolates from 2009. Notably, a recombinant strain emerged in 2015, four years after 4/91 vaccine introduction, indicating vaccine–field-strain genetic exchange. By 2017, isolates with >99% identity to the 4/91 strain were recovered, suggesting vaccine-derived variants. In 2021, GI-1 re-emerged, showing recombination signatures between GI-1 and GI-13 (4/91-derived) strains, likely reflecting suboptimal or inconsistent vaccination strategies. Selection analyses showed strong purifying selection across most of the S1 gene, with limited sites under positive selection in the receptor-binding domain. Phylodynamic reconstruction revealed time-structured evolution and multiple introduction events over 35 years, with lineage-specific tMRCA estimates. Collectively, these findings highlight the emergence of a novel lineage in South America and demonstrate that vaccine use, while mitigating disease, has significantly shaped the evolution of IBV in Chile. Our results underscore the importance of continuous genomic surveillance to inform vaccine strategies and limit recombinant emergence. Full article

Background: Exercise-induced fatigue arises primarily from energy substrate depletion and the accumulation of metabolites such as lactate and ammonia, which impair performance and delay recovery. Emerging evidence implicates gut microbiota modulation—particularly via probiotics—as a means to optimize host energy metabolism and accelerate clearance of fatigue-associated by-products. Objective: This study aimed to determine whether live or heat-inactivated Lacticaseibacillus paracasei NB23 can enhance exercise endurance and attenuate fatigue biomarkers in a murine model. Methods: Forty male Institute of Cancer Research (ICR) mice were randomized into four groups (n = 10 each) receiving daily gavage for six weeks with vehicle, heat-killed NB23 (3 × 10¹⁰ cells/human/day), low-dose live NB23 (1 × 10¹⁰ CFUs/human/day), or high-dose live NB23 (3 × 10¹⁰ CFUs/human/day). Forelimb grip strength and weight-loaded swim-to-exhaustion tests assessed performance. Blood was collected post-exercise to measure serum lactate, ammonia, blood urea nitrogen (BUN), and creatine kinase (CK). Liver and muscle glycogen content was also quantified, and safety was confirmed by clinical-chemistry panels and histological examination. Results: NB23 treatment produced dose-dependent improvements in grip strength (p < 0.01) and swim endurance (p < 0.001). All NB23 groups exhibited significant reductions in post-exercise lactate (p < 0.0001), ammonia (p < 0.001), BUN (p < 0.001), and CK (p < 0.0001). Hepatic and muscle glycogen stores rose by 41–59% and 65–142%, respectively (p < 0.001). No changes in food or water intake, serum clinical-chemistry parameters, or tissue histology were observed. Conclusions: Our findings suggest that both live and heat-treated L. paracasei NB23 may contribute to improved endurance performance, increased energy reserves, and faster clearance of fatigue-related metabolites in our experimental model. However, these results should be interpreted cautiously given the exploratory nature and limitations of our study. Full article