Vaccine Delivery Route Research Articles

Effects of hydrogen peroxidase-inactivated vaccine against Streptococcus agalactiae infection in red tilapia (Oreochromis sp.) via different routes of administration

Hydrogen peroxidase-inactivated vaccines have recently developed in controlling infectious diseases in aquaculture. The present study aimed to compare the efficacy of the hydrogen peroxidase-inactivated Streptococcus agalactiae vaccine via different delivery routes (injection, immersion, and oral administration) in red tilapia (Oreochromis sp.) without using adjuvant. Fish were randomly divided into 4 groups including G1: Control treatment (without vaccine), G2: vaccine-based diet (oral administration), G3: vaccinated by immersion and G4: vaccinated by injection. After 6 weeks of the experiment, fish were intraperitoneally injected with S. agalactiae and the mortality was recorded in 14 days. The results showed that lysozyme activity was differentially increased according to the delivery routes of vaccine, organs and time of sampling. However, the specific antibody levels in all vaccinated groups were only increased in week 6 post-vaccination. After the challenge test with S. agalactiae, the serum lysozyme levels in G3 and G4 were significantly higher than the control group (G1), while the total white blood cells and specific antibody levels were significantly increased in G2 and G4 compared to the control (G1). Similarly, the hydrogen peroxidase-inactivated vaccine statistically reduced the cumulative mortality in G2 (35.29%) and G4 (28.95%) compared to G1 (44.12%) after injected with the S. agalactiae. These results showed that the vaccine delivery routes by oral administration or injection may decrease the pathogen and show better protection for red tilapia than the immersion method. Further studies will be investigated to improve the efficacy of hydrogen peroxidaseinactivated vaccine via using different adjuvants.

Viral-vectored respiratory mucosal vaccine strategies.

Increasing global concerns of pandemic respiratory viruses highlight the importance of developing optimal vaccination strategies thatencompass vaccine platform, delivery route, and regimens. The decades-long effort to develop vaccines to combat respiratory infections such as influenza, respiratory syncytial virus, and tuberculosis has met with challenges, including the inability of systemically administered vaccines to induce respiratory mucosal (RM) immunity. In this regard, ample preclinical and available clinical studies have demonstrated the superiority of RM vaccination to induce RM immunity over parenteral route of vaccination. A great stride has been made in developing vaccines for RM delivery against respiratory pathogens, including M. tuberculosis and SARS-CoV-2. In particular, inhaled aerosol delivery of adenoviral-vectored vaccines has shown significant promise.

The Integrated Consideration of Vaccine Platforms, Adjuvants, and Delivery Routes for Successful Vaccine Development.

Vaccines have proven to be the most cost-efficient and reasonable way to fight and exterminate virulent pathogens. Vaccines can be designed using a variety of platforms including inactivated/attenuated pathogen or subunits of it. The most recent COVID mRNA vaccines have employed nucleic acid sequences for the antigen of interest to combat the pandemic. Different vaccine platforms have been chosen for different licensed vaccines which all have shown their ability to induce durable immune responses and protection. In addition to platforms, different adjuvants have been used to strengthen the immunogenicity of vaccines. Among the delivery routes, intramuscular injection has been the most common for vaccination. In this review, we present a historical overview of the integrated consideration of vaccine platforms, adjuvants, and delivery routes in the success of vaccine development. We also discuss the advantages and limitations of each choice in the efficacy of vaccine development.

A Synthetic SARS-CoV-2-Derived T-Cell and B-Cell Peptide Cocktail Elicits Full Protection against Lethal Omicron BA.1 Infection in H11-K18-hACE2 Mice.

Emerging variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been developing the capacity for immune evasion and resistance to existing vaccines and drugs. To address this, development of vaccines against coronavirus disease 2019 (COVID-19) has focused on universality, strong T cell immunity, and rapid production. Synthetic peptide vaccines, which are inexpensive and quick to produce, show low toxicity, and can be selected from the conserved SARS-CoV-2 proteome, are promising candidates. In this study, we evaluated the effectiveness of a synthetic peptide cocktail containing three murine CD4+ T-cell epitopes from the SARS-CoV-2 nonspike proteome and one B-cell epitope from the Omicron BA.1 receptor-binding domain (RBD), along with aluminum phosphate (Al) adjuvant and 5' cytosine-phosphate-guanine 3' oligodeoxynucleotide (CpG-ODN) adjuvant in mice. The peptide cocktail induced good Th1-biased T-cell responses and effective neutralizing-antibody titers against the Omicron BA.1 variant. Additionally, H11-K18-hACE2 transgenic mice were fully protected against lethal challenge with the BA.1 strain, with a 100% survival rate and reduced pulmonary viral load and pathological lesions. Subcutaneous administration was found to be the superior route for synthetic peptide vaccine delivery. Our findings demonstrate the effectiveness of the peptide cocktail in mice, suggesting the feasibility of synthetic peptide vaccines for humans. IMPORTANCE Current vaccines based on production of neutralizing antibodies fail to prevent the infection and transmission of SARS-CoV-2 Omicron and its subvariants. Understanding the critical factors and avoiding the disadvantages of vaccine strategies are essential for developing a safe and effective COVID-19 vaccine, which would include a more effective and durable cellular response, minimal effects of viral mutations, rapid production against emerging variants, and good safety. Peptide-based vaccines are an excellent alternative because they are inexpensive, quick to produce, and very safe. In addition, human leukocyte antigen T-cell epitopes could be targeted at robust T-cell immunity and selected in the conserved region of the SARS-CoV-2 variants. Our study showed that a synthetic SARS-CoV-2-derived peptide cocktail induced full protection against lethal infection with Omicron BA.1 in H11-K18-hACE2 mice for the first time. This could have implications for the development of effective COVID-19 peptide vaccines for humans.

Intranasal multivalent adenoviral-vectored vaccine protects against replicating and dormant M.tb in conventional and humanized mice

Viral-vectored vaccines are highly amenable for respiratory mucosal delivery as a means of inducing much-needed mucosal immunity at the point of pathogen entry. Unfortunately, current monovalent viral-vectored tuberculosis (TB) vaccine candidates have failed to demonstrate satisfactory clinical protective efficacy. As such, there is a need to develop next-generation viral-vectored TB vaccine strategies which incorporate both vaccine antigen design and delivery route. In this study, we have developed a trivalent chimpanzee adenoviral-vectored vaccine to provide protective immunity against pulmonary TB through targeting antigens linked to the three different growth phases (acute/chronic/dormancy) of Mycobacterium tuberculosis (M.tb) by expressing an acute replication-associated antigen, Ag85A, a chronically expressed virulence-associated antigen, TB10.4, and a dormancy/resuscitation-associated antigen, RpfB. Single-dose respiratory mucosal immunization with our trivalent vaccine induced robust, sustained tissue-resident multifunctional CD4+ and CD8+ T-cell responses within the lung tissues and airways, which were further quantitatively and qualitatively improved following boosting of subcutaneously BCG-primed hosts. Prophylactic and therapeutic immunization with this multivalent trivalent vaccine in conventional BALB/c mice provided significant protection against not only actively replicating M.tb bacilli but also dormant, non-replicating persisters. Importantly, when used as a booster, it also provided marked protection in the highly susceptible C3HeB/FeJ mice, and a single respiratory mucosal inoculation was capable of significant protection in a humanized mouse model. Our findings indicate the great potential of this next-generation TB vaccine strategy and support its further clinical development for both prophylactic and therapeutic applications.

The race toward a universal influenza vaccine: Front runners and the future directions

Influenza virus is the pathogen of influenza (flu) and millions of people suffer from the infection worldwide, posing a significant health risk. The current influenza vaccines induce neutralizing antibodies against hemagglutinin (HA) to achieve strain-specific neutralization. The effectiveness of seasonal vaccines is usually low and unpredictable because of the antigenic variation and genetic plasticity of viruses, as well as the interference of preexisting immunity. A universal influenza vaccine is urgently needed to prevent a wide variety of influenza viruses. Nevertheless, reaching this difficult optimal goal requires a step-by-step approach. Innovative strategies and vaccine platforms are being developed in order to generate robust cross-protective immunity. In this review, we summarize candidate influenza vaccines that meet two criteria: first, they are designed to provide protection against multiple influenza viruses; second, they had passed regulatory evaluations and have entered various stages of clinical trials. We discuss these vaccine candidates based on the different vaccine-production platforms, with the focus on antigen selection, design, adjuvants, immunomodulators, and routes of vaccine delivery in the development of universal influenza vaccines.

Effects of oil-based adjuvants on the immune response of pigs after dermal administration of antigen and evaluation of the immunization level after a subsequent Actinobacillus pleuropneumoniae challenge in pigs

Route of vaccine delivery can greatly impact the immunogenicity, efficacy and safety of the vaccine. Four groups of piglets were immunised transdermally (t.d.), intradermally (i.d.) or intramuscularly (i.m.) with the same doses of antigen in combination with a water-in-oil-in-water emulsion adjuvant Montanide™ ISA 201 VG or with a microemulsion adjuvant Montanide™ IMS 1313 VG N ST (Seppic, France). The last group was left without vaccination as a control group. All animals were subsequently exposed to the infection induced by Actinobacillus pleuropneumoniae (App). The immune response was evaluated with respect to the intensity of systemic and mucosal antibody formation, their isotype characterisation and rate of cell-mediated immunity. These findings were compared with the intensity of adverse local reactions and level of protection in experimental challenge. Monitoring of the local reaction at the injection site after each administration showed that microemulsion adjuvant IMS 1313 was less reactogenic than the water-in-oil-in-water emulsion ISA 201. In terms of efficacy, both dermal administrations were less immunogenic than the i.m route. The i.m. injection induced higher anti-App9 IgG and IgM titres. Nevertheless, IgG1 and IgG2 isotypes analysis revealed a close immunological profile between i.m. and i.d. routes. The concentration of IFN-γ from peripheral blood after in vitro restimulation with the specific antigen was only increased in the i.m. group at the day of challenge (D35) and two weeks after (D49). Interestingly, the smallest gross pulmonary lesions were observed in the i.d. vaccinated group (3.4%) compared to the control group (39.4%) and to groups with other routes of administration. Taken together, these results suggest that i.d. administration of vaccines is a promising approach. Even the i.d. vaccine was more reactogenic and slightly less immunogenic than the i.m. vaccine, its protection effectiveness seemed to be superior.

Comparative immunogenicity analysis of intradermal versus intramuscular immunization with a recombinant human adenovirus type 5 vaccine against Ebola virus.

The proper route for vaccine delivery plays an important role in activating a robust immune response. Several viral vector-based vaccines against Ebola disease administered intramuscularly have been found to have excellent immunogenicity and protectiveness. In this study, we evaluated different vaccine routes for Ad5-EBOV delivery by comparing humoral and cellular responses, germinal center reactions, dendritic cell activation and antigen expression. Mice injected intramuscularly with the vaccine exhibited an advantage in antigen expression, leading to more robust germinal center and humoral responses, while intradermal injection recruited more migrating DCs and induced a more polyfunctional cellular response. Our study provides more data for future use of viral vector-based vaccines.

Differential Biodistribution of Adenoviral-Vectored Vaccine Following Intranasal and Endotracheal Deliveries Leads to Different Immune Outcomes.

Infectious diseases of the respiratory tract are one of the top causes of global morbidity and mortality with lower respiratory tract infections being the fourth leading cause of death. The respiratory mucosal (RM) route of vaccine delivery represents a promising strategy against respiratory infections. Although both intranasal and inhaled aerosol methods have been established for human application, there is a considerable knowledge gap in the relationship of vaccine biodistribution to immune efficacy in the lung. Here, by using a murine model and an adenovirus-vectored model vaccine, we have compared the intranasal and endotracheal delivery methods in their biodistribution, immunogenicity and protective efficacy. We find that compared to intranasal delivery, the deepened and widened biodistribution in the lung following endotracheal delivery is associated with much improved vaccine-mediated immunogenicity and protection against the target pathogen. Our findings thus support further development of inhaled aerosol delivery of vaccines over intranasal delivery for human application.

Development and Testing of a Spray-Dried Tuberculosis Vaccine Candidate in a Mouse Model.

Converting a vaccine into a thermostable dry powder is advantageous as it reduces the resource burden linked with the cold chain and provides flexibility in dosage and administration through different routes. Such a dry powder presentation may be especially useful in the development of a vaccine towards the respiratory infectious disease tuberculosis (TB). This study assesses the immunogenicity and protective efficacy of spray-dried ID93+GLA-SE, a promising TB vaccine candidate, against Mycobacterium tuberculosis (Mtb) in a murine model when administered via different routes. Four administration routes for the spray-dried ID93+GLA-SE were evaluated along with relevant controls—1) reconstitution and intramuscular injection, 2) reconstitution and intranasal delivery, 3) nasal dry powder delivery via inhalation, and 4) pulmonary dry powder delivery via inhalation. Dry powder intranasal and pulmonary delivery was achieved using a custom nose-only inhalation device, and optimization using representative vaccine-free powder demonstrated that approximately 10 and 44% of the maximum possible delivered dose would be delivered for intranasal delivery and pulmonary delivery, respectively. Spray-dried powder was engineered according to the different administration routes including maintaining approximately equivalent delivered doses of ID93 and GLA. Vaccine properties of the different spray-dried lots were assessed for quality control in terms of nanoemulsion droplet diameter, polydispersity index, adjuvant content, and antigen content. Our results using the Mtb mouse challenge model show that both intranasal reconstituted vaccine delivery as well as pulmonary dry powder vaccine delivery resulted in Mtb control in infected mice comparable to traditional intramuscular delivery. Improved protection in these two vaccinated groups over their respective control groups coincided with the presence of cytokine-producing T cell responses. In summary, our results provide novel vaccine formulations and delivery routes that can be harnessed to provide protection against Mtb infection.

The Route of Vaccine Administration Determines Whether Blood Neutrophils Undergo Long-Term Phenotypic Modifications.

Innate immunity modulates adaptive immunity and defines the magnitude, quality, and longevity of antigen-specific T- and B- cell immune memory. Various vaccine and administration factors influence the immune response to vaccination, including the route of vaccine delivery. We studied the dynamics of innate cell responses in blood using a preclinical model of non-human primates immunized with a live attenuated vaccinia virus, a recombinant Modified vaccinia virus Ankara (MVA) expressing a gag-pol-nef fusion of HIV-1, and mass cytometry. We previously showed that it induces a strong, early, and transient innate response, but also late phenotypic modifications of blood myeloid cells after two months when injected subcutaneously. Here, we show that the early innate effector cell responses and plasma inflammatory cytokine profiles differ between subcutaneous and intradermal vaccine injection. Additionally, we show that the intradermal administration fails to induce more highly activated/mature neutrophils long after immunization, in contrast to subcutaneous administration. Different batches of antibodies, staining protocols and generations of mass cytometers were used to generate the two datasets. Mass cytometry data were analyzed in parallel using the same analytical pipeline based on three successive clustering steps, including SPADE, and categorical heatmaps were compared using the Manhattan distance to measure the similarity between cell cluster phenotypes. Overall, we show that the vaccine per se is not sufficient for the late phenotypic modifications of innate myeloid cells, which are evocative of innate immune training. Its route of administration is also crucial, likely by influencing the early innate response, and systemic inflammation, and vaccine biodistribution.

Protection against avian coronavirus conferred by oral vaccination with live bacteria secreting LTB-fused viral proteins

The devastating impact of infectious bronchitis (IB) triggered by the IB virus (IBV), on poultry farms is generally curbed by livestock vaccination with live attenuated or inactivated vaccines. Yet, this approach is challenged by continuously emerging variants and by time limitations of vaccine preparation techniques. This work describes the design and evaluation of an anti-IBV vaccine comprised of E. coli expressing and secreting viral spike 1 subunit (S1) and nucleocapsid N-terminus and C-terminus polypeptides fused to heat-labile enterotoxin B (LTB) (LS1, LNN, LNC, respectively). Following chicken oral vaccination, anti-IBV IgY levels and cellular-mediated immunity as well as protection against virulent IBV challenge, were evaluated 14 days following the booster dose. Oral vaccination induced IgY levels that exceeded those measured following vaccination with each component separately. Following exposure to inactivated IBV, splenocytes isolated from chicks orally vaccinated with LNN or LNC -expressing bacteria, showed a higher percentage of CD8+ cells as compared to splenocytes isolated from chicks vaccinated with wild type or LTB-secreting E. coli and to chicks subcutaneously vaccinated. Significant reduction in viral load and percent of shedders in the vaccinated chicks was evident starting 3 days following challenge with 107.5 EID50/ml virulent IBV. Taken together, orally delivered LTB-fused IBV polypeptide-expressing bacteria induced virus-specific IgY antibody production and was associated with significantly shorter viral shedding on challenge with a live IBV. The proposed vaccine design and delivery route promise an effective and rapidly adaptable means of protecting poultry farms from devastating IB outbreaks.

Live-Attenuated Vaccines Prevent Respiratory Syncytial Virus-Associated Illness in Young Children

To determine the preliminary efficacy and durability of live-attenuated respiratory syncytial virus RSV vaccine candidates from phase 1 clinical trials.Healthy RSV-naïve children aged 6 to 24 months (n = 241) were randomly assigned 2:1 to receive either a live-attenuated RSV vaccine or placebo.Separate trials of 7 different RSV vaccine candidates were similarly conducted, and data were pooled for analysis. The surveillance period was from ∼5.5 to 13 months after day 56 postvaccination. The RSV neutralizing antibody (NeutAb) response, also considered indicative of vaccine efficacy, was defined as more than a fourfold increase in complement-enhanced plaque reduction-neutralizing antibody titers after vaccination. Data were further grouped by “more promising” vaccines (NeutAb response in ≥80% of vaccine recipients; n = 5) and “less promising” (NeutAb response in ≤59% of vaccines; n = 2).There was 66% efficacy against respiratory syncytial virus–associated medically attended acute respiratory illness (RSV-MAARI) infections (P = .007) and 100% efficacy against respiratory syncytial virus–associated medically attended acute lower respiratory illness (RSV-MAALRI) (P = .009) among all NeutAb responders. A NeutAb response overall was significantly associated with protection against RSV-MAARI and RSV-MAALRI (odds ratio: 0.26; 95% confidence interval: 0.09 to 0.75; P = .014). Vaccine virus titers shed by vaccine recipients did not correlate with protection against RSV-MAARI or RSV-MAALRI. Although there was no significant difference in the primary NeutAb responses between the vaccinated and placebo groups (P = .21), the magnitude of anamnestic NeutAb response was ∼10-fold higher in vaccine recipients than in those with the placebo after infection with wild-type RSV during surveillance (P = .0001). In the vaccinated group not infected with wild-type RSV (n = 89), the postsurveillance anamnestic NeutAb titers (79 [6.3 log2]) were less than postimmunization (111 [6.8 log2]; P < .001); however, the absolute difference was small. On the basis of the disease rate model, the authors determined that future study sample sizes between 540 and 1300 would be needed to demonstrate efficacy.Live-attenuated RSV vaccines provide protection against acute respiratory and lower respiratory illnesses and can produce durable response.Currently, palivizumab, a monoclonal antibody, is still the only Food and Drug Administration–approved medication for RSV, providing short-term prophylaxis against severe RSV disease in high-risk infants and children with preexisting conditions, such as prematurity with or without bronchopulmonary dysplasia. The preliminary data from these small clinical trials identifying these “more promising” vaccines suggest that we may be 1 step closer toward actually developing an RSV vaccine that can protect against infection and address the burden of RSV disease. Obvious considerations for future and later stage clinical trials include appropriately powered sample sizes (including RSV-exposed subjects), study duration, geographic locations, timing of the RSV season, and vaccine delivery routes and dosing.

Delivery Routes for COVID-19 Vaccines.

The novel coronavirus, SARS-CoV-2, which causes COVID-19, has resulted in a pandemic with millions of deaths. To eradicate SARS-CoV-2 and prevent further infections, many vaccine candidates have been developed. These vaccines include not only traditional subunit vaccines and attenuated or inactivated viral vaccines but also nucleic acid and viral vector vaccines. In contrast to the diversity in the platform technology, the delivery of vaccines is limited to intramuscular vaccination. Although intramuscular vaccination is safe and effective, mucosal vaccination could improve the local immune responses that block the spread of pathogens. However, a lack of understanding of mucosal immunity combined with the urgent need for a COVID-19 vaccine has resulted in only intramuscular vaccinations. In this review, we summarize the history of vaccines, current progress in COVID-19 vaccine technology, and the status of intranasal COVID-19 vaccines. Future research should determine the most effective route for vaccine delivery based on the platform and determine the mechanisms that underlie the efficacy of different delivery routes.

Early immune responses in skin and lymph node after skin delivery of Toll-like receptor agonists in neonatal and adult pigs

The skin is potentially an important vaccine delivery route facilitated by a high number of resident antigen presenting cells (APCs), which are known to be stimulated by different Toll-like receptor agonists (TLRa). In this study, neonatal and adult pigs were vaccinated in the skin using dissolving microneedle patches to investigate the immuno-stimulatory potential of different TLRa and possible age-dependent differences early after vaccination. These patches contained TLR1/2a (Pam3Cys), TLR7/8a (R848) or TLR9a (CpG ODN) combined with inactivated porcine reproductive and respiratory syndrome virus (PRRSV) or with an oil-in-water stable emulsion. Vaccinated skin and draining lymph nodes were analysed for immune response genes using microfluidic high-throughput qPCR to evaluate the early immune response and activation of APCs. Skin pathology and immunohistochemistry were used to evaluate the local immune responses and APCs in the vaccinated skin, respectively.In both neonatal and adult pigs, skin vaccination with TLR7/8a induced the most prominent early inflammatory and immune cell responses, particularly in the skin. Skin histopathology and immunohistochemistry of APCs showed comparable results for neonatal and adult pigs after vaccination with the different TLRa vaccines. However, in vaccinated neonatal pigs in the skin and draining lymph node more immune response related genes were upregulated compared to adult pigs. We showed that both neonatal and adult skin could be stimulated to develop an immune response, particularly after TLR7/8a vaccination, with age-dependent differences in regulation of immune genes. Therefore, age-dependent differences in local early immune responses should be considered when developing skin vaccines.

Assessment of the Immunogenicity of a Novel Live Recombinant Rift Valley Fever arMP-12andDelta;NSm21/384 Vaccine Candidate Following Intranasal Vaccination of Goats, Sheepand Calves in Tanzania

Background: Rift Valley fever virus (RVFV) is an arbovirus that causes morbidity and mortality in livestock and humans throughout Africa and in the Arabian Peninsula. Vaccines are effective for the prevention of Rift Valley fever (RVF) disease, but new and improved vaccines are needed to improve the safety of available vaccines. Also, non-invasive needle free vaccine delivery routes should be evaluated as an alternative for invasive routes of vaccination. Objective: The aim of this proof of concept study was to evaluate the safety and immunogenicity of a novel live attenuated recombinant RVFV arMP-12ΔNSm21/384 vaccine candidate following intranasal vaccination of goats, calves, and sheep in Tanzania. Methods: Healthy, 6-9 months old breeds of African sheep (Ovis aeris), goats (Capra aegagrus) and zebu calves (Bos taurus indicus) were used in this study. The animals were purchased from local livestock keepers in the Mvomero district of Morogoro region, Tanzania. Animals were seronegative to both RVFV and antibody at the time of use in the vaccine trials. Animals in the test group included 10 goats, 7 sheep and 10 calves that were vaccinated in the left nares with 50 μl each and 2 sheep were vaccinated with 100 μl each (50 μl each in the left and right nares) of a dose that contained 4 × 105 PFU/50 ul of arMP-12ΔNSm21/384 vaccine, while the control group, including 2 goats, 3 sheep and 2 calves that were injected in the left nares with 50 μl of phosphate buffered saline to serve as placebo controls. Rectal temperature was measured and blood samples were collected on day 14 and 0 before vaccination, and on days 3, 5, 7, 14, 21, 28 and 35 post vaccinations (PV). Serum samples collected on days 14 and 0 before vaccination were tested for RVFV neutralizing antibody by a plaque reduction neutralization test, and on days 3 and 5 PV, serum samples were tested for virus as possible evidence of a viremia in cell culture and weekly collected samples thereafter were tested for RVFV neutralizing antibody. Results: All animals were negative for RVFV neutralizing antibody at 14 and 0 days before vaccination and none of the animals had detectable viremia on days 3 and 5 PV, and none had clinical manifestations throughout the study. Among the 7 sheep, 10 goats, and 10 calves that received 50 μl each of the vaccine dose, 70% had the first detectable antibody on either day 5, 7 or 14 PV with titers ranging from 1:10 to 1:40. The 2 sheep that received the 100 μl each of the virus dose had the first detectable antibody on day 5 PV with a titer of 1:160. Subsequently, animals vaccinated with the 50 μl dose had antibody titers ranged from 1:10 to 640 on days 21, 28 and 35 PV, while those vaccinated with 100 μl maintained an antibody titer of 1:160 throughout the study. Moreover, there was no difference in the antibody titers between animal species p=0.34, although mean antibody titers of goats were highest. Conclusion: As a proof of concept studies, the findings demonstrated that intranasal vaccination is a promising route for vaccinating domestic ruminants with the RVFV arMP-12ΔNSm21/384 vaccine candidate. However, these preliminary results suggested that a larger dose of 4 × 105 PFU/100 ul of arMP-12ΔNSm21/384 vaccine needed to be administered to each animal to consistently elicit a robust immune response. Also, further studies are warranted using a larger number of domestic ruminants to confirm the immune response elicited by the larger dose of the vaccine administered via the intranasal route.

Vaccination Route Determines the Kinetics and Magnitude of Nasal Innate Immune Responses in Rainbow Trout (Oncorhynchus mykiss).

Simple SummaryMany pathogens exploit the olfactory route to reach critical organs in the body such as the brain or lungs. Thus, effective onset of an early innate immune response in the nasal epithelium is key to stopping pathogen progression. The stimulation of nasal immunity by vaccines may depend on the type of vaccine and vaccination route. The goal of this study was to evaluate the ability of a live attenuated viral vaccine to stimulate innate immunity in the olfactory organ of rainbow trout, a teleost fish of commercial aquaculture value. The kinetics and magnitude of the innate immune response depended on the route of vaccination, with the strongest and fastest responses recorded in intranasally vaccinated fish. Injection vaccination had an intermediate effect, whereas immersion vaccination resulted in delayed and weak nasal innate immunity. Injection vaccination, even with the vehicle control, induced mortality in fingerlings, whereas nasal and immersion vaccines were safe. Challenge experiments with the live virus revealed that nasal and injected vaccines conferred very high and comparable levels of protection, but immersion vaccination only induced transient protection. In conclusion, the route of vaccination determines the type, magnitude and velocity of the innate immune response in the nasal epithelium of animals.Many pathogens infect animal hosts via the nasal route. Thus, understanding how vaccination stimulates early nasal immune responses is critical for animal and human health. Vaccination is the most effective method to prevent disease outbreaks in farmed fish. Nasal vaccination induces strong innate and adaptive immune responses in rainbow trout and was shown to be highly effective against infectious hematopoietic necrosis (IHN). However, direct comparisons between intranasal, injection and immersion vaccination routes have not been conducted in any fish species. Moreover, whether injection or immersion routes induce nasal innate immune responses is unknown. The goal of this study is to compare the effects of three different vaccine delivery routes, including intranasal (IN), intramuscular (i.m.) injection and immersion (imm) routes on the trout nasal innate immune response. Expression analyses of 13 immune-related genes in trout nasopharynx-associated lymphoid tissue (NALT), detected significant changes in immune expression in all genes analyzed in response to the three vaccination routes. However, nasal vaccination induced the strongest and fastest changes in innate immune gene expression compared to the other two routes. Challenge experiments 7 days post-vaccination (dpv) show the highest survival rates in the IN- and imm-vaccinated groups. However, survival rates in the imm group were significantly lower than the IN- and i.m.-vaccinated groups 28 dpv. Our results confirm that nasal vaccination of rainbow trout with live attenuated IHNV is highly effective and that the protection conferred by immersion vaccination is transient. These results also demonstrate for the first time that immersion vaccines stimulate NALT immune responses in salmonids.

Construction of a Live-Attenuated Vaccine Strain of Yersinia pestis EV76-B-SHUΔpla and Evaluation of Its Protection Efficacy in a Mouse Model by Aerosolized Intratracheal Inoculation.

Plague, which is caused by Yersinia pestis, is one of the most dangerous infectious diseases. No FDA-approved vaccine against plague is available for human use at present. To improve the immune safety of Y. pestis EV76 based live attenuated vaccine and to explore the feasibility of aerosolized intratracheal inoculation (i.t.) route for vaccine delivery, a plasminogen activator protease (pla) gene deletion mutant of the attenuated Y. pestis strain EV76-B-SHU was constructed, and its residual virulence and protective efficacy were evaluated in a mouse model via aerosolized intratracheal inoculation (i.t.) or via subcutaneous injection (s.c.). The residual virulence of EV76-B-SHUΔpla was significantly reduced compared to that of the parental strain EV76-B-SHU following i.t. and s.c. infection. The EV76-B-SHUΔpla induced higher levels of mucosal antibody sIgA in the bronchoalveolar lavage fluid of mice immunized by i.t. but not by s.c.. Moreover, after lethal challenge with Y. pestis biovar Microtus strain 201 (avirulent in humans), the protective efficacy and bacterial clearance ability of the EV76-B-SHUΔpla-i.t. group were comparable to those of the EV76-B-SHUΔpla-s.c. and EV76-B-SHU immunized groups. Thus, the EV76-B-SHUΔpla represents an excellent live-attenuated vaccine candidate against pneumonic plague and aerosolized i.t. represents a promising immunization route in mouse model.

Lipids as Activators of Innate Immunity in Peptide Vaccine Delivery.

Innate immune system plays an important role in pathogen detection and the recognition of vaccines, mainly through pattern recognition receptors (PRRs) that identify pathogen components (danger signals). One of the typically recognised bacterial components are lipids in conjugation with peptides, proteins and saccharides. Lipidic compounds are readily recognised by the immune system, and thus are ideal candidates for peptide- based vaccine delivery. Thus, bacterial or synthetic lipids mixed with, or conjugated to, antigens have shown adjuvant properties. These systems have many advantages over traditional adjuvants, including low toxicity and good efficacy for stimulating mucosal and systemic immune responses. The most recent literature on the role of lipids in stimulation of immune responses was selected for this review. The vast majority of reviewed papers were published in the last decade. Older but significant findings are also cited. This review focuses on the development of lipopeptide vaccine systems including application of palmitic acid, bacterial lipopeptides, glycolipids and the lipid core peptide and their routes of administration. The use of liposomes as a delivery system that incorporates lipopeptides is discussed. The review also includes a brief description of immune system in relation to vaccinology and discussion on vaccine delivery routes. Lipids and their conjugates are an ideal frontrunner in the development of safe and efficient vaccines for different immunisation routes.

Oral Immunization with HIV-1 Envelope SOSIP trimers elicits systemic immune responses and cross-reactive anti-V1V2 antibodies in non-human primates.

Development of a successful HIV vaccine is dependent upon a determination of the optimum antigen and adjuvant as well as choosing an optimal site for vaccine delivery. The site of delivery is particularly relevant as HIV transmission generally requires that the virus crosses a mucosal membrane to infect a new host. Here we undertake a pilot study comparing three vaccine delivery routes, two to the oral cavity (intraepithelial (iEp) and needle-free (NF-Injex)) as well as intramuscular (IM) delivery. These vaccinations utilized a recombinant HIV-1 Env trimer 10042.05 from an elite neutralizer, subject VC10042, that has previously induced high titers of cross-clade reactive V1V2 antibodies. The 10042.05.SOSIP fused trimer was administered with adjuvants R848 (Resiquimod), MPLA and Alhydrogel to characterize the innate cellular and anti-HIV Envelope (Env) antibody responses following the administration of the vaccine to the oral mucosa. Oral delivery of the 10042.05.SOSIP induced high titers of anti-V1V2 antibodies, which together with previous studies, indicates an immunogenic bias toward the V1V2 regions in 10042-derived Envs. Both types of oral vaccine delivery resulted in immunologic and serologic responses that were comparable to the IM delivery route. Furthermore, induction of anti-V1-V2 specific antibodies was best following iEp delivery of the oral vaccine identifying this as the optimal method to orally deliver this vaccine formulation.