Immune response after an experimental intramammary challenge with killed Staphylococcus aureus in cows and heifers vaccinated and not vaccinated with Startvac, a polyvalent mastitis vaccine.

An experimental trial was conducted to explore the effect of vaccination with a polyvalent vaccine against mastitis (Startvac) on the early immune response after experimental intramammary challenge with a heterologous killed Staphylococcus aureus strain. The effect of vaccination on milk production, clinical signs, quarter milk somatic cell count, milk polymorphonuclear neutrophilic leukocyte (PMN) concentration and viability, the concentration of antigen-specific antibodies [slime associated antigenic complex (SAAC) and J5] and their IgG1 and IgG2 subtypes in both serum and whey, and the antigen-specific IFN-γ, IL-4, and IL-17 production by blood lymphocytes after in vitro stimulation with S. aureus and Escherichia coli extracts were determined. A cohort of 8 clinically healthy end-term cows and heifers were conveniently selected, of which half was vaccinated with Startvac at 45 and 10 d before the expected calving date and half served as nonvaccinated control animals. At 15 d in milk, 2 contralateral quarters of each of the 8 animals were challenged with 2×109 cfu/mL of the formaldehyde-killed S. aureusC195strain. The 2 other quarters were infused with phosphate-buffered saline and served as control quarters. The increase in both quarter milk somatic cell count and PMN concentration and the drop in milk production after S. aureus inoculation was less pronounced in the vaccinates than in the nonvaccinates, reflecting a less severe inflammatory response. No significant differences in PMN viability between vaccinates and nonvaccinates could be demonstrated. The serum SAAC- and J5-specific antibody concentration significantly increased across the dry period in the vaccinated animals only. The whey concentration of SAAC-specific antibodies was significantly higher in vaccinates than in nonvaccinates at both 15 and 17 d in milk, independent from the challenge status of the quarters. No significant differences in the whey J5-specific antibody concentration were observed. Vaccination with Startvac seems to primarily evoke a Th2 response for S. aureus characterized by a shift toward the IgG1 antibody subtype and accompanied by a less pronounced Th1 response. The type of response against E. coli was less clear, though a weak but significant shift toward the IgG2 antibody subtype after vaccination and high IFN-γ levels after in vitro stimulation suggest a Th1 response. The increased SAAC-specific antibody concentration in whey in vaccinates compared with nonvaccinates most probably triggers the opsonization of the inoculated S. aureus bacteria, resulting in a more efficient elimination of the bacteria from the mammary gland.

Immunogenicity and safety in pigs of PHH-1V, a SARS-CoV-2 RBD fusion heterodimer vaccine candidate.

The continuing high global incidence of COVID-19 and the undervaccinated status of billions of persons strongly motivate the development of a new generation of efficacious vaccines. We have developed an adjuvanted vaccine candidate, PHH-1V, based on a protein comprising the receptor binding domain (RBD) of the Beta variant of SARS-CoV-2 fused in tandem with the equivalent domain of the Alpha variant, with its immunogenicity, safety and efficacy previously demonstrated in mouse models. In the present study, we immunized pigs with different doses of PHH-1V in a prime-and-boost scheme showing PHH-1V to exhibit an excellent safety profile in pigs and to produce a solid RBD-specific humoral response with neutralising antibodies to 7 distinct SARS-CoV-2 variants of concern, with the induction of a significant IFNγ+ T-cell response. We conclude that PHH-1V is safe and elicits a robust immune response to SARS-CoV-2 in pigs, a large animal preclinical model.

Preclinical evaluation of PHH-1V vaccine candidate against SARS-CoV-2 in non-human primates.

SARS-CoV-2 emerged in December 2019 and quickly spread worldwide, continuously striking with an unpredictable evolution. Despite the success in vaccine production and mass vaccination programs, the situation is not still completely controlled, and therefore accessible second-generation vaccines are required to mitigate the pandemic. We previously developed an adjuvanted vaccine candidate coded PHH-1V, based on a heterodimer fusion protein comprising the RBD domain of two SARS-CoV-2 variants. Here, we report data on the efficacy, safety, and immunogenicity of PHH-1V in cynomolgus macaques. PHH-1V prime-boost vaccination induces high levels of RBD-specific IgG binding and neutralizing antibodies against several SARS-CoV-2 variants, as well as a balanced Th1/Th2 cellular immune response. Remarkably, PHH-1V vaccination prevents SARS-CoV-2 replication in the lower respiratory tract and significantly reduces viral load in the upper respiratory tract after an experimental infection. These results highlight the potential use of the PHH-1V vaccine in humans, currently undergoing Phase III clinical trials.

Preclinical evaluation of a COVID-19 vaccine candidate based on a recombinant RBD fusion heterodimer of SARS-CoV-2.

Current COVID-19 vaccines have been associated with a decline in infection rates, prevention of severe disease, and a decrease in mortality rates. However, SARS-CoV-2 variants are continuously evolving, and development of new accessible COVID-19 vaccines is essential to mitigate the pandemic. Here, we present data on preclinical studies in mice of a receptor-binding domain (RBD)-based recombinant protein vaccine (PHH-1V) consisting of an RBD fusion heterodimer comprising the B.1.351 and B.1.1.7 SARS-CoV-2 variants formulated in SQBA adjuvant, an oil-in-water emulsion. A prime-boost immunisation with PHH-1V in BALB/c and K18-hACE2 mice induced a CD4+ and CD8+ T cell response and RBD-binding antibodies with neutralizing activity against several variants, and also showed a good tolerability profile. Significantly, RBD fusion heterodimer vaccination conferred 100% efficacy, preventing mortality in SARS-CoV-2 infected K18-hACE2 mice, but also reducing Beta, Delta and Omicron infection in lower respiratory airways. These findings demonstrate the feasibility of this recombinant vaccine strategy.

Efficacy of a Salmonella enterica serovar Abortusovis (S. Abortusovis) inactivated vaccine in experimentally infected gestating ewes.

Salmonella enterica serovar Abortusovis (S. Abortusovis) infection is one of the most important causes of infectious late-term abortion as well as birth of weak lambs in sheep in many countries throughout the world. Implementation of protocols based on the application of effective vaccines is one of the most effective approaches for controlling this disease, but variable efficacy has been reported, possibly related to factors associated with the host, the vaccine, the parameters used for determining efficacy and the challenge protocols. In this context, a new commercial inactivated vaccine (INMEVA; Laboratorios Hipra S.A., Spain) was evaluated in 20 control and 17 vaccinated gestating ewes, subcutaneously challenged at 90 days of gestation with 5 × 106 colony-forming units (cfu) of a wild strain of S. Abortusovis. Incidence of reproductive failures, bacterial vaginal excretion (by real time PCR), and lamb survival were evaluated as indicators of the vaccine's level of protection. Moreover, humoral response (by ELISA test in serum samples) was studied. Vaccination was showed to be safe under the study conditions. Vaccine efficacy was demonstrated in two different ways: i) it significantly decreased the percentage of abortions [29.4% (5/17) in the vaccinated group compared to the control group (65%; 13/20)] and ii) there was a significant reduction of the overall vaginal excretion in the sampling period (3.05 log cfu/mL ±â€¯0.84 in the vaccinated group vs. 5.68 ±â€¯0.67 in the control group). Given these results, the vaccine evaluated can be considered as an effective alternative for controlling S. Abortusovis infection in ovine flocks.

NASYM, a live intranasal vaccine, protects young calves from bovine respiratory syncytial virus in the presence of maternal antibodies.

BACKGROUND: Bovine respiratory syncytial virus (BRSV) is a major problem for cattle worldwide during their first year of life. The aim of the present study was to evaluate efficacy and longevity of immunity of a live vaccine (NASYM, HIPRA) in the presence of maternally derived antibodies (MDA). METHOD: Calves (36) were distributed in four groups, based on MDA status and treatment. They received NASYM or a placebo at an early age (less than two weeks) by intranasal route. Eight weeks later, animals were challenged with the Asquith strain of BRSV. Efficacy was assessed by monitoring clinical signs and mortality, PaO2 , virus shedding and lung lesions. The immunological response was evaluated by measuring IgG in serum and IgA in nasal secretions. RESULTS: A reduction of mortality, lung lesions, shedding and a higher PaO2 was achieved in NASYM vaccinated groups, independently of MDA status. An anamnestic IgG response was observed after challenge in vaccinated animals, both in MDA+ and MDA- groups. An IgA response was also observed in vaccinated animals after vaccination and challenge. CONCLUSION: NASYM protected newborn calves with MDAs during the first 10 weeks of life, against a very virulent challenge that caused extensive pulmonary lesions and deaths in control animals, with just a single intranasal dose.

Evaluation of the Efficacy of a New Commercially Available Inactivated Vaccine Against Ovine Enzootic Abortion.

Ovine enzootic abortion (OEA), caused by Chlamydia abortus, is an economically important disease in many countries. Inactivated vaccines have been used for many years as they induce immunity in sheep, although outbreaks of abortions have been described in vaccinated flocks. In addition, there is a commercially available live attenuated vaccine that provides good protective results. Recently however, reports question the attenuation of this vaccine and associate it with the appearance of outbreaks of OEA in vaccinated flocks. In the present study, a recently commercialized inactivated vaccine (INMEVA®; Laboratorios Hipra S.A., Amer, Spain) has been evaluated using mouse and sheep experimental models. In the mouse models (non-pregnant and pregnant models), the efficacy of INMEVA vaccine has been compared to an unvaccinated control group and to an experimental inactivated vaccine considered as a positive protection control (UMU vaccine). In the non- pregnant model, the UMU vaccine was more effective than the INMEVA vaccine regarding the impact on body weight or the presence of C. abortus in the liver, but both vaccinated groups (UMU and INMEVA) had significantly lower C. abortus in the liver compared to the control group. In the pregnant model in terms of reproductive failures, pups per mouse or the presence of C. abortus in the liver or uterus, no significant differences were found between both vaccines, inducing protection compared to the control group. In the ovine pregnant model, where INMEVA vaccine was compared only to an unvaccinated group, the results indicate that this new commercial vaccine is safe and provides a suitable level of protection against an experimental challenge with C. abortus. A 75% reduction in reproductive disorders, 55% reduction in animals with C. abortus shedding on day of parturition/abortion, and a significant reduction of the average amount of chlamydial shedding from parturition/abortion over the next 21 days was observed, in relation to the infected control group. The results suggest that this vaccine is adequate for the control and prevention of OEA; however, future studies are necessary to elucidate the type of protective immune response that it induces.

Study of the humoral immunological response after vaccination with a Staphylococcus aureus biofilm-embedded bacterin in dairy cows: possible role of the exopolysaccharide specific antibody production in the protection from Staphylococcus aureus induced mastitis.

The objective of the present study was to analyze an extracellular component from Staphylococcus aureus (S. aureus), which we refer to as slime associated antigenic complex (SAAC), and to investigate the role of SAAC-specific antibody production in protection from S. aureus bovine mastitis. Twelve primiparous gestating cows were randomly assigned to one of the three groups: Group 1 was vaccinated with a S. aureus bacterin with very limited SAAC content; Group 2 received a S. aureus bacterin with high SAAC content and Group 3 served as unvaccinated controls. Animals were vaccinated at 45 days before the expected parturition date and revaccinated 35 days later. All groups were challenged by intramammary infusion with a virulent heterologous strain of S. aureus 23 days after calving. Antibody response against SAAC in serum and in milk, general clinical signs, mastitis score, somatic cell count (SCC) and count of S. aureus in milk were evaluated before and after challenge. Immunization with a high SAAC content in the S. aureus bacterin (Group 2) significantly enhanced antibody titers against SAAC (in serum and milk) and reduced the S. aureus concentration in milk during the post-challenge period compared to Group 1 and Group 3. Moreover, a significant negative correlation was observed between SAAC antibody production on the day of the challenge and the S. aureus count in milk by 1 day after challenge. However, there was no evidence of a difference between vaccinated and control groups with regard to clinical signs of mastitis following the challenge. Nevertheless, the SAAC antibody concentration on the day of the challenge negatively correlated with the mastitis score in quarters infected with S. aureus at 2 days post-challenge. These results indicate that the vaccines did not prevent S. aureus intramammary infection (IMI) after the experimental challenge, but immunization with a S. aureus bacterin with high SAAC content was able to reduce S. aureus multiplication in the mammary gland after challenge and suggests that the SAAC-specific antibody response could be involved in the protection against S. aureus intramammary infection. Although further studies should be performed to confirm the efficacy (under experimental conditions and in field trials), we propose bacterins from strong biofilm-producing bacteria and with high SAAC content, rather than with limited SAAC content, as a cost-efficient vaccine design against S. aureus bovine mastitis.

Reactivity of 2-pyridinecarboxylic esters with cadmium(II) halides: study of (113)Cd NMR solid state spectra and crystal structures of hexacoordinated complexes [CdI(2)(C(5)H(4)NCOOMe)(2)] and [CdI(2)(C(5)H(4)NCOOPr(n))(2)].

The series of complexes [CdX(2)(C(5)H(4)NCOOR)] (X = Cl or Br; R = Me, Et, Pr(n)() or Pr(i)()) and [CdX(2)(C(5)H(4)NCOOR)(2)] (X = I; R = Me, Et, Pr(n)(), or Pr(i)()) have been obtained by the addition reaction of esters of 2-pyridinecarboxylic acid to cadmium(II) halides. X-ray crystal structures of two complexes [CdI(2)(C(5)H(4)NCOOR)(2)], R = Me (10) and R = Pr(n)() (12), have been determined. In both cases, the structure consists of discrete neutral monomeric units where the cadmium atom has a distorted octahedral coordination with CdI(2)N(2)O(2) core, two halides being in cis disposition. Structural information is compared with that deduced from (113)Cd CPMAS NMR experiments. Chemical shift anisotropies are discussed in terms of distortions produced in cadmium octahedra. The orientation of the principal axes of (113)Cd shielding tensor is also analyzed and related to the disposition of ligands in the structures of two analyzed compounds.