Flagellin Enhances the Immunogenicity of Pasteurella multocida Lipoprotein E Subunit Vaccine.

Fowl cholera, caused by Pasteurella multocida infection, poses challenges for prevention because of its many serotypes. Bacterins are currently widely used for vaccination against fowl cholera, but protection is limited to homologous strains. Live attenuated vaccines of P. multocida provide some heterologous protection, but side effects are considerable. More recently, protein-based antigens are promising subunit vaccines when their low immunogenicity has been addressed with effective adjuvants. Bacterial flagellin has been widely considered a promising adjuvant for vaccines. In this study, we tested the adjutancy of flagellin in a subunit vaccine against P. multocida in a mice and chicken models. For vaccine formulation, the antigen fPlpE (P. multocida liporotein E) was combined with fFliC (Salmonella Typhimurium flagellin). The recombinant proteins of fPlpE and fFliC were successfully expressed using the Escherichia coli system as the expected sizes of 55 kDa and 70 kDa, respectively. The fFliC elicited strong expression levels of proinflammatory cytokine (IL-1ß, IL-8, and IL-6) when stimulated in native chicken peripheral blood mononuclear cells. Immunization of mice and chickens with the subunit vaccines containing fFliC accelerated the antibody response. In the challenge tests, fFliC increased vaccine protective efficacy against the heterologous strain P. multocida A1 and highly virulent strain Chu01 in mice and chickens, respectively. These data indicated potential possibilities of using fFliC as an immunostimulant adjuvant in developing a subunit vaccine against fowl cholera.

Nota de Investigación- La flagelina aumenta la inmunogenicidad de una vacuna subunitaria que contiene lipoprote'ina E de Pasteurella multocida. La enfermedad del cólera aviar, causada por la infección por Pasteurella multocida, representa desaf'ios para la prevención debido a sus numerosos serotipos. Actualmente, las bacterinas se utilizan ampliamente para la vacunación contra el cólera aviar, pero la protección se limita a cepas homólogas. Las vacunas vivas atenuadas de P. multocida proporcionan cierta protección heteróloga, pero los efectos secundarios son considerables. Recientemente, el uso de ant'igenos basados en prote'inas parece ser prometedor como vacunas subunitarias cuando se ha abordado el problema de su baja inmunogenicidad con adyuvantes eficaces. La flagelina bacteriana se ha considerado ampliamente como un adyuvante prometedor para las vacunas. En este estudio, se analizó la capacidad como adyuvante de la flagelina en una vacuna de subunitaria contra P. multocida en modelos de ratones y pollos. Para la formulación de la vacuna, el ant'igeno fPlpE (liporote'ina E de P. multocida) se combinó con fFliC (flagelina de Salmonella Typhimurium). Las prote'inas recombinantes de fPlpE y fFliC se expresaron con éxito utilizando el sistema de E. coli con los tamaños esperados de 55 kDa y 70 kDa, respectivamente. El fFliC indujo fuertes niveles de expresión de citoquinas proinflamatorias (IL-1ß, IL-8 e IL-6) cuando se estimuló en células mononucleares de sangre periférica de pollos nativos. La inmunización de ratones y pollos con las vacunas subunitarias que contienen fFliC aceleró la respuesta de anticuerpos. En las pruebas de desaf'io, la fFliC aumentó la eficacia protectora de la vacuna contra la cepa heteróloga de P. multocida A1 y la cepa altamente virulenta Chu01 en ratones y pollos, respectivamente. Estos datos indicaron posibilidades potenciales de utilizar fFliC como adyuvante inmunoestimulante en el desarrollo de una vacuna subunitaria contra el cólera aviar.

Trivalent outer membrane vesicles-based combination vaccine candidate induces protective immunity against Campylobacter and invasive non-typhoidal Salmonella in adult mice.

Campylobacter and invasive non-typhoidal Salmonella (iNTS) are among the most common causative agents of gastroenteritis worldwide. As of now, no single combination licensed vaccine is available for public health use against both iNTS and Campylobacter species. Outer-membrane vesicles (OMVs) are nanoscale proteoliposomes released from the surface of gram-negative bacteria during log phase and harbor a variety of immunogenic proteins. Based on epidemiology of infections, we formulated a novel trivalent outer membrane vesicles (TOMVs)-based vaccine candidate against Campylobacter jejuni (CJ), Salmonella Typhimurium (ST) and Salmonella Enteritidis (SE). Isolated OMVs from CJ, ST and SE were combined in equal ratios for formulation of TOMVs and 5 µg of the developed vaccine candidate was used for intraperitoneal immunization of adult BALB/c mice. Immunization with TOMVs significantly activated both the humoral and cellular arm of adaptive immune response. Robust bactericidal effect was elicited by TOMVs immunized adult mice sera. TOMVs immunization induced long-term protective efficacy against CJ, ST and SE infections in mice. The study illustrates the ability of TOMVs-based combination immunogen in eliciting broad-spectrum protective immunity against prevalent Campylobacter and iNTS pathogens. According to the findings, TOMVs can work as a potent combination-based acellular vaccine candidate for amelioration of Campylobacter and iNTS-mediated gastroenteritis.

Evaluation of the phase-specific antibody response in water buffalo (Bubalus bubalis) after two doses of an inactivated phase I Coxiella burnetii vaccine.

The control and management of Q fever outbreaks in ruminants are currently based on vaccination. Although buffalo (Bubalus bubalis) are intensively farmed in several countries and represent a reservoir for Coxiellosis, no evidence has been described regarding the efficacy of vaccination in this species. This work aimed to evaluate the humoral response, using appropriate phase-specific ELISAs, and the effects on abortion rate in buffalo by a field study. A total of 15 seropositive and 20 seronegative animals were vaccinated twice, three weeks apart, with a commercial phase I vaccine, and phase-specific antibodies were determined in the course of vaccination. Although anti-phase II antibody reactivity predominated after vaccination compared to phase I, both anti-phase I- and -phase II-antibody-reactivity significantly increased after the first (p = 0.001) and again after the second vaccination (p = 0.05). Seroconversion did not significantly depend on age or natural infection status. Once the vaccination cycle was completed, the herd study observed a reduced rate of abortion and placenta retention. Our data demonstrated that the vaccine principally induced a similar antibody response as in goats and sheep. These preliminary data appeared to support vaccination in buffalo, even in seropositive animals, although further studies are needed to better define the dynamics concerning seroconversion in this species.

A multivalent mRNA-LNP vaccine protects against Clostridioides difficile infection.

Clostridioides difficile infection (CDI) is an urgent public health threat with limited preventative options. In this work, we developed a messenger RNA (mRNA)-lipid nanoparticle (LNP) vaccine targeting C. difficile toxins and virulence factors. This multivalent vaccine elicited robust and long-lived systemic and mucosal antigen-specific humoral and cellular immune responses across animal models, independent of changes to the intestinal microbiota. Vaccination protected mice from lethal CDI in both primary and recurrent infection models, and inclusion of non-toxin cellular and spore antigens improved decolonization of toxigenic C. difficile from the gastrointestinal tract. Our studies demonstrate mRNA-LNP vaccine technology as a promising platform for the development of novel C. difficile therapeutics with potential for limiting acute disease and promoting bacterial decolonization.

Field efficacy of a recombinant toxoid vaccine against Shiga toxin 2e during a naturally occurring edema disease infection.

The objective of this field trial was to determine the efficacy of a recombinant toxoid vaccine against Shiga toxin 2e (Stx2e) in piglets suffering from edema disease (ED). Three farms with confirmed ED cases were selected for the field trials. On each farm, a total of 40 4-day-old pigs were randomly allocated to either the vaccinated or unvaccinated group, with 20 pigs per group (10 males and 10 females). A 1.0-mL dose of the recombinant toxoid vaccine was administered intramuscularly to pigs in the vaccinated groups in accordance with the manufacturer's recommendations at 4 d of age. A single 2.0-mL dose of phosphate-buffered saline was administered to unvaccinated pigs at the same age. Clinical signs of ED were observed in 12 piglets in the unvaccinated groups and 7 unvaccinated pigs died as a result of ED out of the total number of piglets on Farms A, B, and C. Vaccination had a positive effect on pig growth performance compared to that of unvaccinated pigs on 2 of the 3 farms. Seroconversion of neutralizing antibodies against Stx2e occurred in 70% of piglets in the vaccinated group on Farm A, 75% of vaccinated piglets on Farm B, and 55% of vaccinated piglets on Farm C, when detectable antibodies were measured at 17 d post-vaccination (dpv). Detectable antibodies were measured in 85% of vaccinated piglets on Farms A and B and in 90% of these piglets on Farm C at 37 dpv. These field trials determined that the ED recombinant toxoid vaccine successfully reduced clinical signs and mortality, improved average daily weight gain, and resulted in the production of neutralizing antibodies against Stx2e in pigs.

L'objectif de cette étude sur le terrain était de déterminer l'efficacité d'un vaccin à base d'anatoxine recombinante contre la toxine Shiga 2e (Stx2e) chez les porcelets souffrant de la maladie de l'oedème (ED). Trois fermes ayant des cas confirmés de ED ont été sélectionnées pour les études sur le terrain. Dans chaque ferme, un total de 40 porcs âgés de 4 jours ont été répartis au hasard dans le groupe vacciné ou non vacciné, avec 20 porcs par groupe (10 mâles et 10 femelles). Une dose de 1,0 ml du vaccin à base d'anatoxine recombinante a été administrée par voie intramusculaire aux porcs des groupes vaccinés conformément aux recommandations du fabricant à l'âge de 4 jours. Une dose unique de 2,0 ml de solution saline tamponnée au phosphate a été administrée aux porcs non vaccinés au même âge. Des signes cliniques de ED ont été observés chez 12 porcelets des groupes non vaccinés et 7 porcs non vaccinés sont morts des suites d'une maladie de l'oedème sur le nombre total de porcelets des fermes A, B et C. La vaccination a eu un effet positif sur les performances de croissance des porcs par rapport à celles des porcs non vaccinés dans 2 des 3 fermes. La séroconversion des anticorps neutralisants contre Stx2e s'est produite chez 70 % des porcelets du groupe vacciné de la ferme A, 75 % des porcelets vaccinés de la ferme B et 55 % des porcelets vaccinés de la ferme C, lorsque des anticorps détectables ont été mesurés 17 jours après la vaccination (dpv). Des anticorps détectables ont été mesurés chez 85 % des porcelets vaccinés des fermes A et B et chez 90 % de ces porcelets de la ferme C à 37 dpv. Ces études sur le terrain ont déterminé que le vaccin toxoïde recombinant ED réduisait avec succès les signes cliniques et la mortalité, améliorait le gain de poids quotidien moyen et entraînait la production d'anticorps neutralisants contre Stx2e chez les porcs.(Traduit par Docteur Serge Messier).

Protection of Broiler Chickens Against Necrotic Enteritis by Intrapulmonary Delivery of a Live Clostridium perfringens Vaccine Exploiting the Gut-Lung-Axis Concept.

Clostridium perfringens (CP)-induced necrotic enteritis (NE) is an economically important disease in the broiler chicken industry. The incidence of NE is common in 3-to-6-wk-old broiler chickens, once maternal antibodies start declining. Developing an effective vaccination strategy against NE, preferably delivering a single dose of vaccine at hatch to protect broiler chickens against NE without a booster vaccine, is an enormous challenge. The objective of this study was to induce mucosal immunity in the intestines against NE by intrapulmonary (IPL) delivery of a live CP vaccine at hatch, exploiting the gut-lung-axis (GLA) concept by vaccine delivery following in ovo administration of cytosine-phosphorothioate-guanine oligodeoxynucleotides (CpG-ODN) to induce immune cell maturation in the lungs. Experiments were conducted to explore the dose of CP and immune protection against heterologous CP challenge, and to study the efficacy of IPL delivery of a CP vaccine without a booster. Additional studies were conducted to measure serum immunoglobulin (Ig)Y, mucosal IgA, and histopathology of lungs following vaccination. Delivery of a live CP vaccine by the IPL route, with or without in ovo CpG-ODN, provided significant protection against NE (P < 0.0001). Systemic IgY and mucosal IgA against CP were correlated with protection against NE. There was no necrosis or inflammation in the pulmonary parenchyma. There was a low number of CP isolated from the lungs following live CP delivery by the IPL route. A significant influx of (P < 0.001) of CD8+ T cells and macrophages were noted in the lungs 2 days following live CP delivery by the IPL route. IPL delivery of a live CP vaccine, rather than inactivated CP, provided better protection. This study demonstrated the utility in exploiting the GLA concept in vaccine delivery in broiler chickens.

Protección de pollos de engorde contra la enteritis necrótica mediante la administración intrapulmonar de una vacuna viva de Clostridium perfringens que aprovecha el concepto del eje intestino-pulmón. La enteritis necrótica (NE) inducida por Clostridium perfringens (CP) es una enfermedad económicamente importante en la industria de los pollos de engorde. La incidencia de enteritis necrótica es común en pollos de engorde de tres a seis semanas de edad, una vez que los anticuerpos maternos comienzan a disminuir. El desarrollo de una estrategia de vacunación eficaz contra la enteritis necrótica, preferiblemente administrando una dosis única de vacuna en la eclosión para proteger a los pollos de engorde contra dicha enfermedad sin la necesidad de aplicar una vacuna de refuerzo, es un desafío importante. El objetivo de este estudio fue inducir inmunidad de mucosas en los intestinos contra la enteritis necrótica mediante la administración intrapulmonar (IPL) de una vacuna de C. perfringens viva en la eclosión, explotando el concepto del eje intestino-pulmón (GLA) mediante la administración de la vacuna después de la administración in ovo de citosin oligodesoxinucleótidos de fosforotioato-guanina (CpG-ODN) para inducir la maduración de las células inmunitarias en los pulmones. Se realizaron experimentos para explorar la dosis de C. perfringens y la protección inmune contra la exposición heteróloga a C. perfringens, y para estudiar la eficacia de la administración intrapulmonar de una vacuna C. perfringens sin refuerzo. Se realizaron estudios adicionales para medir la inmunoglobulina (Ig)Y sérica, la IgA de mucosas y la histopatología de los pulmones después de la vacunación. La administración de una vacuna de C. perfringens viva por vía intrapulmonar, con o sin CpG-ODN in ovo, proporcionó una protección significativa contra la enteritis necrótica (P < 0.0001). La IgY sistémica y la IgA mucosa contra C. perfringens se correlacionaron con la protección contra la enteritis necrótica. No hubo necrosis ni inflamación en el parénquima pulmonar. Hubo un número bajo de C. perfringens aislado de los pulmones después de la administración de la vacuna viva contra C. perfringens por vía intrapulmonar. Se observó una afluencia significativa (P < 0.001) de células T CD8+ y macrófagos en los pulmones dos días después de la administración de C. perfringens viva por vía intrapulmonar. La administración mediante vía intrapulmonar de una vacuna viva contra C. perfringens, en lugar de la vacuna contra C. perfringens inactivada, proporcionó una mejor protección. Este estudio demostró la utilidad de explotar el concepto del eje intestino-pulmón en la administración de vacunas en pollos de engorde.

Stability characterizations of feed-based bivalent vaccine containing inactivated Streptococcus agalactiae and Aeromonas hydrophila against streptococcosis and Aeromonas infections in red hybrid tilapia (Oreochromis sp.).

Feed-based bivalent vaccine (FBBV) containing killed whole organism (KWO) of Streptococcus agalactiae and Aeromonas hydrophila with 10% palm oil was previously proved to improve red hybrid tilapia's (Oreochromis sp.) immunity against streptococcosis and Aeromonas infections. This study characterized the FBBV's stability following the preparatory process and storage. The FBBV was prepared, and the KWO's stability was determined microscopically and molecularly. The efficacy of FBBV stored at room temperature (25 ± 2 °C) for 0, 30 and 60 days was investigated in red hybrid tilapia. The results indicated the addition of palm oil was not affecting the KWO's structure and helping in the FBBV's pelletization. In 1 g of FBBV contained 1.5 × 109 CFU/g of S. agalactiae and 4.9 × 109 CFU/g of A. hydrophila, respectively, even after 60 days of storage at room temperature. The KWO's structure in FBBV was not affected following in vitro acidic tolerance analysis, as noted from light and electron microscopies. The FBBV's carbohydrate, energy, moisture, total protein and total ash contents remained stable at 95% after 60 days of storage at room temperature, while the KWO's concentration was slightly reduced to 83.3% for S. agalactiae (1.25 × 109 CFU/g) and 80.6% for A. hydrophila (3.85 × 109 CFU/g), respectively. Fish vaccinated with FBBV that was stored for 0, 30 and 60 days did not show any significant differences (p ≥ 0.05) in the relative percent survival when challenged with pathogenic Streptococcus spp. and Aeromonas spp. These findings suggested that the FBBV is a stable vaccine, which underscores its potential application as aquatic vaccines in aquaculture.

Protection of Rabbits Against Colonization and Morbidity Associated With Toxigenic Pasteurella multocida by Immunization With Inactivated Heat-labile Toxin.

BACKGROUND/AIM: Pasteurella multocida is a significant cause of morbidity and mortality in rabbits, as well as other species. Some isolates elaborate a heat-labile toxin (PMT) that has been shown to be an important virulence factor. Though previous studies have demonstrated protective immunity can be conferred via immunization of rabbits with heat-inactivated PMT (IPMT), we investigated the ability of immunization to impact colonization of P. multocida. MATERIALS AND METHODS: Rabbits were immunized at days 0, 7 and 14 with either phosphate buffered saline (PBS), the mucosal adjuvant cholera toxin (CT), IMPT or IPMT + CT. Male New Zealand white rabbits were used and confirmed to be free of P. multocida prior to experimentation. RESULTS: Serum IgG and nasal lavage fluid IgA responses directed against PMT were found in rabbits immunized with IPMT, with or without CT, but not in those immunized with only PBS or CT; and the addition of CT to IPMT enhanced the response. Significantly more P. multocida CFUs (p≤0.05) were cultured from the lungs of rabbits immunized with IPMT, with or without CT, compared to those administered only PBS or CT, although no differences were observed in nasal lavage fluid samples. Further, immunization IPMT, with or without CT, conferred protection against pleuritis and pneumonia. CONCLUSION: PMT, in addition to its role as a virulence factor, may serve as a colonization factor for P. multocida in the lungs of rabbits.

Enhancement of clinical signs in C3H/HeJ mice vaccinated with a highly immunogenic Leptospira methyl-accepting chemotaxis protein following challenge.

Leptospirosis is the most widespread zoonosis and a life-threatening disease in humans and animals. Licensed killed whole-cell vaccines are available for animals; however, they do not offer heterologous protection, do not induce long-term protection, or prevent renal colonization. In this study, we characterized an immunogenic Leptospira methyl-accepting chemotaxis protein (MCP) identified through a reverse vaccinology approach, predicted its structure, and tested the protective efficacy of a recombinant MCP fragment in the C3H/HeJ mice model. The predicted structure of the full-length MCP revealed an architecture typical for topology class I MCPs. A single dose of MCP vaccine elicited a significant IgG antibody response in immunized mice compared to controls (P < 0.0001), especially the IgG1 and IgG2a subclasses. The vaccination with MCP, despite eliciting a robust immune response, did not protect mice from disease and renal colonization. However, survival curves significantly differed between groups, and the MCP-vaccinated group developed clinical signs faster than the control group. There were differences in gross and histopathological changes between the MCP-vaccinated and control groups. The factors leading to enhanced disease process in vaccinated animals need further investigation. We speculate that anti-MCP antibodies may block the MCP signaling cascade and may limit chemotaxis, preventing Leptospira from reaching its destination, but facilitating its maintenance and replication in the blood stream. Such a phenomenon may exist in endemic areas where humans are highly exposed to Leptospira antigens, and the presence of antibodies might lead to disease enhancement. The role of this protein in Leptospira pathogenesis should be further evaluated to comprehend the lack of protection and potential exacerbation of the disease process. The absence of immune correlates of protection from Leptospira infection is still a major limitation of this field and efforts to gather this knowledge are needed.

The current status and development forecasts of vaccines for aquaculture and its effects on bacterial and viral diseases.

The aquaculture sector predicts protein-rich meals by 2040 and has experienced significant economic shifts since 2000. However, challenges emanating from disease control measures, brood stock improvement, feed advancements, hatchery technology, and water quality management due to environmental fluctuations have been taken as major causative agents for hindering the sector's growth. For the past years, aquatic disease prevention and control have principally depended on the use of various antibiotics, ecologically integrated control, other immunoprophylaxis mechanisms, and chemical drugs, but the long-term use of chemicals such as antibiotics not only escalates antibiotic-resistant bacteria and genes but also harms the fish and the environments, resulting in drug residues in aquatic products, severely obstructing the growth of the aquaculture sector. The field of science has opened new avenues in basic and applied research for creating and producing innovative and effective vaccines and the enhancement of current vaccines to protect against numerous infectious diseases. Recent advances in vaccines and vaccinology could lead to novel vaccine candidates that can tackle fish diseases, including parasitic organism agents, for which the current vaccinations are inadequate. In this review, we study and evaluate the growing aquaculture production by focusing on the current knowledge, recent progress, and prospects related to vaccinations and immunizations in the aquaculture industry and their effects on treating bacterial and viral diseases. The subject matter covers a variety of vaccines, such as conventional inactivated and attenuated vaccines as well as advanced vaccines, and examines their importance in real-world aquaculture scenarios. To encourage enhanced importation of vaccines for aquaculture sustainability and profitability and also help in dealing with challenges emanating from diseases, national and international scientific and policy initiatives need to be informed about the fundamental understanding of vaccines.

Optimization and evaluation of a chitosan-coated PLGA nanocarrier for mucosal delivery of Porphyromonas gingivalis antigens.

Recent advances in understanding Alzheimer's disease (AD) suggest the possibility of an infectious etiology, with Porphyromonas gingivalis emerging as a prime suspect in contributing to AD. P. gingivalis may invade systemic circulation via weakened oral/intestinal barriers and then cross the blood-brain barrier (BBB), reaching the brain and precipitating AD pathology. Based on the proposed links between P. gingivalis and AD, a prospective approach is the development of an oral nanovaccine containing P. gingivalis antigens for mucosal delivery. Targeting the gut-associated lymphoid tissue (GALT), the nanovaccine may elicit both mucosal and systemic immunity, thereby hampering P. gingivalis ability to breach the oral/intestinal barriers and the BBB, respectively. The present study describes the optimization, characterization, and in vitro evaluation of a candidate chitosan-coated poly(lactic-co-glycolic acid) (PLGA-CS) nanovaccine containing a P. gingivalis antigen extract. The nanocarrier was prepared using the double emulsion solvent evaporation method and optimized for selected experimental factors, e.g. PLGA amount, surfactant concentration, w1/o phase ratio, applying a d-optimal statistical design to target the desired physicochemical criteria for its intended application. After nanocarrier optimization, the nanovaccine was characterized in terms of particle size, polydispersity index (PdI), ζ-potential, encapsulation efficiency (EE), drug loading (DL), morphology, and in vitro release profile, as well as for mucoadhesivity, stability under simulated gastrointestinal conditions, antigen integrity, in vitro cytotoxicity and uptake using THP-1 macrophages. The candidate PLGA-CS nanovaccine demonstrated appropriate physicochemical, mucoadhesive, and antigen release properties for oral delivery, along with acceptable levels of EE (55.3 ± 3.5 %) and DL (1.84 ± 0.12 %). The integrity of the encapsulated antigens remained uncompromised throughout NPs production and simulated gastrointestinal exposure, as confirmed by SDS-PAGE and Western blotting analyses. Furthermore, the nanovaccine showed effective in vitro uptake, while exhibiting low cytotoxicity. Taken together, these findings underscore the potential of PLGA-CS NPs as carriers for adequate antigen mucosal delivery, paving the way for further investigations into their applicability as vaccine candidates against P. gingivalis.

Syphilis vaccine development: Aligning vaccine design with manufacturing requirements.

Syphilis, caused by Treponema pallidum subsp. pallidum, is a global health concern with increasing rates worldwide. Current prevention strategies, including screen-and-treat approaches, are not sufficient to resolve rising infection rates, emphasizing the need for a vaccine. Developing a syphilis vaccine necessitates a range of cross-disciplinary considerations, including essential disease-specific protection, technical requirements, economic feasibility, manufacturing constraints, public acceptance, equitable vaccine access, alignment with global public vaccination programs, and identification of essential populations to be vaccinated to achieve herd immunity. Central to syphilis vaccine development is prioritization of global vaccine availability, including access in low- to middle-income settings. Various vaccine platforms, including subunit, virus-like particle (VLP), mRNA, and outer membrane vesicle (OMV) vaccines, present both advantages and challenges. The proactive consideration of both manufacturing feasibility and efficacy throughout the pre-clinical research and development stages is essential for producing an efficacious, inexpensive, and scalable syphilis vaccine to address the growing global health burden caused by this disease.

Preferences for a Clostridioides difficile vaccine among adults in the United States.

INTRODUCTION: Clostridioides difficile (C.diff) infection (CDI) causes significant morbidity and mortality among older adults. Vaccines to prevent CDI are in development; however, data on the target population's preferences are needed to inform vaccination recommendations in the United States (US). This study assessed US adults' willingness to receive a C.diff vaccine and examined how vaccine attributes influence their choices. METHODS: A cross-sectional online survey with a discrete choice experiment (DCE) was conducted among US adults aged ≥50 years. DCE attributes included effectiveness, duration of protection, reduction in symptom severity, out-of-pocket (OOP) costs, number of doses, and side effects. The DCE included 11 choice tasks, each with two hypothetical vaccine profiles and an opt-out (i.e., no vaccine). Attribute-level preference weights were estimated using hierarchical Bayesian modeling. Attribute relative importance (RI) was compared between select subgroups. RESULTS: Of 1216 adults in the analyses, 29.9% reported they knew either 'a little' (20.7%) or 'a lot' (9.2%) about C.diff before the study. A C.diff vaccine was chosen 58.0% of the time (vs. opt-out) across choice tasks. It was estimated that up to 75.0% would choose a vaccine when OOP was $0. Those who were immunocompromised/high-risk for CDI (vs. not) more frequently chose a C.diff vaccine. Decreases in OOP costs (RI = 56.1), improvements in vaccine effectiveness (RI = 17.7), and reduction in symptom severity (RI = 10.3) were most important to vaccine choice. The rank ordering of attributes by importance was consistent across subgroups. CONCLUSION: OOP cost, improvements in vaccine effectiveness, and reduction in CDI severity were highly influential to vaccine selection. Most adults aged ≥50 years were receptive to a C.diff vaccine, especially with little-to-no OOP cost, suggesting that mandating insurance coverage of vaccination with no copayment may increase uptake. The limited awareness about C.diff among adults presents an opportunity for healthcare providers to educate their patients about CDI prevention.

Evaluation of virulence of Aeromonas veronii strain GZ21-2 and development of a highly effective vaccine for grass carp with the potential for industrial application.

Bacterial septicemia represents a significant disease affecting cultured grass carp culture, with the primary etiological agent identified as the Gram-negative bacterium Aeromonas veronii. In response to an outbreak of septicemia in Guangzhou, we developed a formaldehyde-inactivated vaccine against an A. veronii strain designated AV-GZ21-2. This strain exhibited high pathogenicity in experimental infections across at all developmental stages of grass carp. Mortality rates for grass carp weighing 15 ± 5 g ranged from 16 % to 92 % at exposure temperatures of 19 °C-34 °C, respectively. The median lethal dose (LD50) for grass carp groups weighing 15 ± 5 g, 60 ± 10 g, 150 ± 30 g and 500 ± 50 g were determined to be 1.43, 2.52, 4.65 and 7.12 × 107(CFU/mL), respectively. We investigated the inactivated vaccine in conbination with aluminum hydroxide gel (AV-AHG), Montanide ISA201VG (AV-201VG), and white oil (AV-WO) adjuvants. This study aimed to optimize inactivation conditions and identify the adjuvant that elicits the most robust immune response. The AV-GZ21-2 inactivated bacterial solution (AV),when combined with various adjuvants, was capable of inducing a strong specific immune response in grass carp. The relative percent survival (RPS) following a lethal challenge with AV-GZ21-2 were 94 % for AV-AHG, 88 % for AV-201VG, 84 % for AV-WO and 78 % for AV alone. The minimum immunization dose of the AV-AHG vaccine was determined to be 6.0 × 107 CFU per fish, providing immunity for a duration of six months with an immune protection level exceeding 75 %. Furthermore, the AV-AHG vaccine demonstrated significant protective efficacy against various epidemic isolates of A. veronii. Consequently, we developed an inactivated vaccine targeting a highly pathogenic strain of A. veronii, incorporating an aluminum hydroxide gel adjuvant, which resulted in high immune protection and a duration of immunity exceeding six months. These findings suggest that the AV-AHG vaccine holds substantial potential for industrial application.

Immune response and safety of co-administered peste des petits ruminants, contagious caprine pleuropneumonia, sheep and goat pox, and Pasteurellosis vaccines in goats.

Background: Infectious diseases such as peste des petits ruminants (PPRs), contagious caprine pleuropneumonia (CCPP), sheep and goat pox (SGPX), and pasteurellosis have considerable impacts on the optimal utilization of sheep and goat resources in Ethiopia. Immunization using multiple vaccines administered simultaneously has been suggested as a cost-effective and safe approach to controlling and preventing these diseases. Aim: The aim of this study was to assess the immunogenicity and safety of multiple vaccines administered simultaneously in goats. Methods: Sero-negative PPR, CCPP, SGPX, and Pasteurellosis goats were immunized with multiple vaccines. Goats vaccinated with a single vaccine against each disease served as a positive control. The immune response of the goats was assessed using serological tests, and any adverse effects were monitored. Results: The results of the present study showed that goats vaccinated with multiple vaccines exhibited a remarkable immune response against PPR, CCPP, and pasteurellosis. In contrast, they did not produce a protective immune response against sheep or goat pox. No adverse effects were observed with any of the vaccines. Conclusion: This study suggested that combined vaccines can be effective at inducing a protective immune response in goats. However, further research is needed to fully understand the immune response to combined vaccines.

Development of novel multi-protein chimeric immunogens that protect against infection with the Lyme disease agent, Borreliella burgdorferi.

Lyme disease is the most common tick-borne disease in North America. A vaccine for use in humans is not available. Here, we detail the development of two chimeric vaccine antigens, BAF and Chv2M. BAF elicits Abs that target proteins and protein variants produced by Borreliella species in ticks (OspB and OspA) and mammals (FtlA/B). OspB serves as the backbone structure for the BAF chimeric. Two OspA221-240 epitope-containing domain (ECD) variants (#A1 and #A15) were inserted into a loop in OspB. The N-terminal region of the FtlA protein was joined to the C-terminus of the chimeric. The second chimeric, Chv2M, consists of L5 (loop 5) and H5 (helix 5) ECDs derived from diverse OspC proteins. Borreliella species produce OspC upon exposure to the bloodmeal and during early infection in mammals. Here, we demonstrate that BAF and Chv2M are potent immunogens that elicit Abs that bind to each component protein (FtlA, FtlB, OspB, and multiple OspA and OspC variants). Anti-BAF and anti-Chv2M Abs kill Borreliella burgdorferi strains through Ab-mediated complement-dependent and complement-independent mechanisms. Eighty percent (32/40) of mice that received a three-dose vaccine regimen were protected from infection with B. burgdorferi B31. Efficacy increased to 90% (18/20) when the amount of Chv2M was increased in the third vaccine dose. Readouts for infection were flaB PCR and seroconversion to VlsE. This study establishes proof of principle for a chimeric immunogen vaccine formulation that elicits Abs to multiple targets on the B. burgdorferi cell surface produced during tick and mammalian stages of the enzootic cycle.IMPORTANCELyme disease is a growing public health threat across parts of the Northern Hemisphere. Regions that can support sustained tick populations are expanding, and the incidence of tick-borne diseases is increasing. In light of the increasing risk of Lyme disease, effective preventive strategies are needed. Most vaccine development efforts have focused on outer surface protein A, a Borreliella burgdorferi protein produced only in ticks. Herein, we describe the development of a novel vaccine formulation consisting of two multivalent chimeric proteins that are immunogenic and elicit antibodies with bactericidal activity that target several cell surface proteins produced by the Lyme disease spirochetes in feeding ticks and mammals. In a broader sense, this study advances efforts to develop custom-designed vaccinogens comprised of epitope-containing domains from multiple proteins.

Novel recombinant vaccinia virus-vectored vaccine affords complete protection against homologous Borrelia burgdorferi infection in mice.

The rising prevalence of Lyme disease (LD) in North America and Europe has emerged as a pressing public health concern. Despite the availability of veterinary LD vaccines, no vaccine is currently available for human use. Outer surface protein C (OspC) found on the outer membrane of the causative agent, Borrelia burgdorferi, has been identified as a promising target for LD vaccine development due to its sustained expression during mammalian infection. However, the efficacy and immunological mechanisms of LD vaccines solely targeting OspC are not well characterized. In this study, we developed an attenuated Vaccinia virus (VV) vectored vaccine encoding type A OspC (VV-OspC-A). Two doses of the VV-OspC-A vaccine conferred complete protection against homologous B. burgdorferi challenge in mice. Furthermore, the candidate vaccine also prevented the development of carditis and lymph node hyperplasia associated with LD. When investigating the humoral immune response to vaccination, VV-OspC-A was found to induce a robust antibody response predominated by the IgG2a subtype, indicating a Th1-bias. Using a novel quantitative flow cytometry assay, we also determined that elicited antibodies were capable of inducing antibody-dependent cellular phagocytosis in vitro. Finally, we demonstrated that VV-OspC-A vaccination generated a strong antigen-specific CD4+ T-cell response characterized by the secretion of numerous cytokines upon stimulation of splenocytes with OspC peptides. This study suggests a promising avenue for LD vaccine development utilizing viral vectors targeting OspC and provides insights into the immunological mechanisms that confer protection against B. burgdorferi infection.

Oral administration of Lactobacillus plantarum expressing aCD11c modulates cellular immunity alleviating inflammatory injury due to Klebsiella pneumoniae infection.

BACKGROUND: Klebsiella pneumoniae (KP), responsible for acute lung injury (ALI) and inflammation of the gastrointestinal tract, is a zoonotic pathogen that poses a threat to livestock farming worldwide. Nevertheless, there is currently no validated vaccine to prevent KP infection. The development of mucosal vaccines against KP using Lactobacillus plantarum (L. plantarum) is an effective strategy. RESULTS: Firstly, the L. plantarum strains NC8-pSIP409-aCD11c' and NC8-pLc23-aCD11c were constructed via homologous recombination to express the aCD11c protein either inducibly or constitutively. Both NC8-pSIP409-aCD11c' and NC8-pLc23-aCD11c strains could enhance the adhesion and invasion of L. plantarum on bone marrow-derived dendritic cells (BMDCs), and stimulate the activation of BMDCs compared to the control strain NC8-pSIP409 in vitro. Following oral immunization of mice with NC8-pSIP409-aCD11c' and NC8-pLc23-aCD11c, the cellular, humoral, and mucosal immunity were significantly improved, as evidenced by the increased expression of CD4+ IL-4+ T cells in the spleen, IgG in serum, and secretory IgA (sIgA) in the intestinal lavage fluid (ILF). Furthermore, the protective effects of L. plantarum against inflammatory damage caused by KP infection were confirmed by assessing the bacterial loads in various tissues, lung wet/dry ratio (W/D), levels of inflammatory cytokines, and histological evaluation, which influenced T helper 17 (Th17) and regulatory T (Treg) cells in peripheral blood and lung. CONCLUSIONS: Both the inducible and constitutive L. plantarum strains NC8-pSIP409-aCD11c' and NC8-pLc23-aCD11c have been found to stimulate cellular and humoral immunity levels and alleviate the inflammatory response caused by KP infection. These findings have provided a basis for the development of a novel vaccine against KP.

Clostridium perfringens antigens and challenges for development of vaccines against necrotic enteritis in poultry.

INTRODUCTION: Chickens with Necrotic Enteritis (NE), caused by Clostridium perfringens, exhibit acute and chronic symptoms that are difficult to diagnose, leading to significant economic losses. Vaccination is the best method for controlling and preventing NE. However, only two vaccines based on the CPA and NetB toxins have been commercialized, offering partial protection, highlighting the urgent need for more effective vaccines. OBJECTIVE: This review aimed to identify promising antigens for NE vaccine formulation and discuss factors affecting their effectiveness. METHODS: A systematic review using five scientific databases identified 30 eligible studies through the Rayyan tool, which were included for quality review. RESULTS: We identified 25 promising antigens, including CPA, NetB, FBA, ZMP, CnaA, FimA, and FimB, categorized by their role in disease pathogenesis. This review discusses the biochemical, physiological, and genetic traits of recombinant antigens used in vaccine prototypes, their expression systems, and immunization potential in chickens challenged with virulent C. perfringens strains. Market supply challenges, immunogenic potential, vaccine platforms, adjuvants, and factors related to vaccination schedules-such as administration routes, dosing intervals, and age at immunization-are also addressed. Additionally, the study notes that vaccine formulations tested under mild challenges may not offer adequate field-level protection due to issues replicating aggressive conditions, strain virulence loss, and varied methodologies. CONCLUSIONS: An ideal NE vaccine should incorporate multiple antigens, molecular adjuvants, and delivery systems via in ovo and oral routes. The review underscores the challenges in developing and validating NE vaccines and the urgent need for a standardized protocol to replicate aggressive challenges accurately.