Safety and immunogenicity of inactivated Rift Valley Fever Smithburn viral vaccine in sheep.

BACKGROUND: The live-attenuated Rift Valley Fever Smithburn (SB) vaccine is one of the oldest products widely used in ruminants for control of RVF infections. Vaccinations with RVF Smithburn result in residual pathogenic effect and is limited for use in non-pregnant animals. Commercially available RVFV inactivated vaccines are considered safer options to control the disease. These products are prepared from virulent RVFV isolates and present occupational safety concerns. This research study evaluates the ability of an inactivated SB vaccine strain to elicit neutralising antibody response in sheep. METHODS: The RVF Smithburn vaccine was inactivated with binary ethylenimine at 37 °C. Inactivated RVFV cultures were adjuvanted with Montande™ Gel-01 and aluminium hydroxide (Al (OH)3) gel for immunogenicity and safety determination in sheep. The commercial RVF inactivated vaccine and a placebo were included as positive and negative control groups, respectively. RESULTS: Inactivated RVFV vaccine formulations were safe with all animals showing no clinical signs of RVFV infection and temperature reactions following prime-boost injections. The aluminium hydroxide formulated vaccine induced an immune response as early as 14 days post primary vaccination with neutralising antibody titre of 1:20 and a peak antibody titre of 1:83 was reached on day 56. A similar trend was observed in the animal group vaccinated with the commercial inactivated RVF vaccine obtaining the highest antibody titre of 1:128 on day 56. The neutralizing antibody levels remained within a threshold for the duration of the study. Merino sheep vaccinated with Montanide™ Gel-01-Smithburn were characterised with overall lower immune response when compared to aluminium hydroxide vaccine emulsions. CONCLUSIONS: These finding suggests that the inactivated RVF Smithburn vaccine strain adjuvanted with aluminium-hydroxide can be used an alternative to the products prepared from virulent RVFV isolates for protection of ruminants against the disease. The vaccine can further be evaluated for safety in pregnant ewes.

Production of recombinant lumpy skin disease virus A27L and L1R proteins for application in diagnostics and vaccine development.

Vaccination using live attenuated vaccines (LAVs) is considered the most effective method for control of lumpy skin disease (LSD). However, this method is limited by safety concerns, with reports of adverse reactions following vaccination. This study evaluates A27L and L1R which are essential proteins for virus attachment and membrane fusion as recombinant sub-unit vaccines against LSD. These proteins were recombinantly expressed in Escherichia coli and purified using affinity chromatography. Purified proteins were formulated individually (A27L or L1R) and in combination (A27L and L1R) with 10% (w/w) Montanide™ Gel 01 PR adjuvant at a final antigen dose of 20 µg per protein. The safety and immunogenicity of these formulations were evaluated in rabbits in a 42-day clinical trial. Animals were vaccinated on day 0 and boost injection administered 21 days later. No reduced morbidity, increased temperature and any other clinical signs were recorded in vaccinated animals for all three vaccine formulations. The highest neutralizing antibody response was detected on day 42 post-primary vaccination for all formulations when using serum neutralising assay. The neutralisation data correlates with antibody titres quantified using a whole cell ELISA. Evaluating the combination of A27L and L1R as potential diagnostic reagents showed highest sensitivity for detection of antibodies against LSD when compared to individual proteins. This study reports the immunogenicity of recombinant A27L and L1R combination for successful application in LSD vaccine development. Furthermore, these proteins demonstrated the potential use in LSD diagnostics.

Improved safety profile of inactivated Neethling strain of the Lumpy Skin Disease Vaccine.

The Lumpy Skin Disease Virus (LSDV) Neethling vaccine strains have been used for decades for prophylactic immunization of domestic ruminants against the disease. Commercial products against Lumpy skin disease are supplied as live attenuated vaccines and often are associated with adverse reactions warranting studies towards development of safe and efficacious vaccine alternatives. The present study was designed to investigate the ability of Montanide™ Gel 01 PR adjuvanted inactivated Neethling vaccine strain of the lumpy skin disease to induce immune response in rabbits. Complete virus inactivation was achieved following treatment of live vaccine strain with binary ethyleneimine (BEI) at 2 mM final concentration. Inactivated virus antigen, formulated with Montanide™ Gel 01 was injected at 1,00E + 05 and 1,00E + 06 TCID50 per dose in rabbits. The second injection with same vaccine dosages was administered 21 days after the primary vaccination. Rabbits that received a 1,00E + 05 TCID50/dose of inactivated LSDV vaccine formulation induced maximum neutralizing antibody titres on day 13 post second vaccinations. Rabbits vaccinated and prime boosted with the 1,00E + 06 TCID50/dose of inactivated LSDV vaccine formulation, induced neutralizing antibody titres on day 14 after first vaccination. The maximum antibody titres for the 1,00E + 06 TCID50/dose of the inactivated LSDV vaccine formulation was obtained on day 35 post vaccination. The 1,00E + 06 TCID50 dose of the inactivated LSDV vaccine Montanide™ Gel-01 PR formulation induced higher neutralizing antibodies. The MontanideTM Gel-01 PR offers safer profile to oil adjuvants and can be developed further to protect target animals against LSDV in non-endemic areas.