Rift Valley Fever Virus Vaccine Research Articles

Evaluation of a Combined Live Attenuated Vaccine against Lumpy Skin Disease, Contagious Bovine Pleuropneumonia and Rift Valley Fever.

The use of effective vaccines is among the most important strategies for the prevention and progressive control of transboundary infectious animal diseases. However, the use of vaccine is often impeded by the cost, a lack of cold chains and other factors. In resource-limited countries in Africa, one approach to improve coverage and reduce cost is to vaccinate against multiple diseases using combined vaccines. Therefore, the objective of this study was to evaluate a combined vaccine for the prevention and control of Lumpy Skin Disease (LSD), Contagious Bovine Pleuropneumonia (CBPP) and Rift Valley fever (RVF). The LSD and CBPP were formulated as a combined vaccine, and the RVF was formulated separately as live attenuated vaccines. These consisted of a Mycoplasma MmmSC T1/44 strain that was propagated in Hayflick-modified medium, RVF virus vaccine, C13T strain prepared in African green monkey cells (Vero), and the LSDV Neethling vaccine strain prepared in primary testis cells. The vaccines were tested for safety via the subcutaneous route in both young calves and pregnant heifers with no side effect, abortion or teratogenicity. The vaccination of calves induced seroconversions for all three vaccines starting from day 7 post-vaccination (PV), with rates of 50% for LSD, 70% for CBPP and 100% for RVF, or rates similar to those obtained with monovalent vaccines. The challenge of cattle vaccinated with the LSD/CBPP and the RVF vaccine afforded full protection against virulent strains of LSDV and RVFV. A satisfactory level of protection against a CBPP challenge was observed, with 50% of protection at 6 months and 81% at 13 months PV. A mass vaccination trial was performed in four regions of Burkina Faso that confirmed safety and specific antibody responses induced by the vaccines. The multivalent LSD/CBPP+RVF vaccine provides a novel and beneficial approach to the control of the three diseases through one intervention and, therefore, reduces the cost and improves vaccination coverage.

Immunoinformatics approach to Rift Valley fever virus vaccine design in ruminants

Immunoinformatics approach to Rift Valley fever virus vaccine design in ruminants

A Novel Quadruple Gene-Deleted BoHV-1-Vectored RVFV Subunit Vaccine Induces Humoral and Cell-Mediated Immune Response against Rift Valley Fever in Calves.

Rift Valley fever virus (RVFV) is considered to be a high biodefense priority based on its threat to livestock and its ability to cause human hemorrhagic fever. RVFV-infected livestock are also a significant risk factor for human infection by direct contact with contaminated blood, tissues, and aborted fetal materials. Therefore, livestock vaccination in the affected regions has the direct dual benefit and one-health approach of protecting the lives of millions of animals and eliminating the risk of severe and sometimes lethal human Rift Valley fever (RVF) disease. Recently, we have developed a bovine herpesvirus type 1 (BoHV-1) quadruple gene mutant virus (BoHV-1qmv) vector that lacks virulence and immunosuppressive properties due to the deletion of envelope proteins UL49.5, glycoprotein G (gG), gE cytoplasmic tail, and US9 coding sequences. In the current study, we engineered the BoHV-1qmv further by incorporating a chimeric gene sequence to express a proteolytically cleavable polyprotein: RVFV envelope proteins Gn ectodomain sequence fused with bovine granulocyte-macrophage colony-stimulating factor (GMCSF) and Gc, resulting in a live BoHV-1qmv-vectored subunit vaccine against RVFV for livestock. In vitro, the resulting recombinant virus, BoHV-1qmv Sub-RVFV, was replicated in cell culture with high titers. The chimeric Gn-GMCSF and Gc proteins expressed by the vaccine virus formed the Gn-Gc complex. In calves, the BoHV-1qmv Sub-RVFV vaccination was safe and induced moderate levels of the RVFV vaccine strain, MP12-specific neutralizing antibody titers. Additionally, the peripheral blood mononuclear cells from the vaccinated calves had six-fold increased levels of interferon-gamma transcription compared with that of the BoHV-1qmv (vector)-vaccinated calves when stimulated with heat-inactivated MP12 antigen in vitro. Based on these findings, we believe that a single dose of BoHV-1qmv Sub-RVFV vaccine generated a protective RVFV-MP12-specific humoral and cellular immune response. Therefore, the BoHV-1qmv sub-RVFV can potentially be a protective subunit vaccine for cattle against RVFV.

An adenovirus-vectored RVF vaccine confers complete protection against lethal RVFV challenge in A129 mice.

Instruction: Rift valley fever virus (RVFV) is a mosquito-transmitted bunyavirus that causes severe disease in animals and humans. Nevertheless, there are no vaccines applied to prevent RVFV infection for human at present. Therefore, it is necessary to develop a safe and effective RVFV vaccine. Methods: We generated Ad5-GnGcopt, a replication-deficient recombinant Ad5 vector (human adenovirus serotype 5) expressing codon-optimized RVFV glycoproteins Gn and Gc, and evaluated its immunogenicity and protective efficacy in mice. Results and Discussion: Intramuscular immunization of Ad5-GnGcopt in mice induces strong and durable antibody production and robust cellular immune responses. Additionally, a single vaccination with Ad5-GnGcopt vaccination can completely protect interferon-α/β receptor-deficient A129 mice from lethal RVFV infection. Our work indicates that Ad5-GnGcopt might represent a potential vaccine candidate against RVFV. However, further research is needed, first to confirm its efficacy in a natural animal host, and ultimately escalate as a potential vaccine candidate for humans.

Immunogenicity of a Candidate Live Attenuated Vaccine for Rift Valley Fever Virus with a Two-Segmented Genome.

Rift Valley fever virus (RVFV) is an emerging arbovirus that affects both ruminants and humans. RVFV causes severe and recurrent outbreaks in Africa and the Arabian Peninsula with a significant risk for emergence into new locations. Although there are a variety of RVFV veterinary vaccines for use in endemic areas, there is currently no licensed vaccine for human use; therefore, there is a need to develop and assess new vaccines. Herein, we report a live-attenuated recombinant vaccine candidate for RVFV, based on the previously described genomic reconfiguration of the conditionally licensed MP12 vaccine. There are two general strategies used to develop live-attenuated RVFV vaccines, one being serial passage of wild-type RVFV strains to select attenuated mutants such as Smithburn, Clone 13, and MP12 vaccine strains. The second strategy has utilized reverse genetics to attenuate RVFV strains by introducing deletions or insertions within the viral genome. The novel candidate vaccine characterized in this report contains a two-segmented genome that lacks the medium viral segment (M) and two virulence genes (nonstructural small and nonstructural medium). The vaccine candidate, named r2segMP12, was evaluated for the production of neutralizing antibodies to RVFV in outbred CD-1 mice. The immune response induced by the r2segMP12 vaccine candidate was directly compared to the immune response induced by the rMP12 parental strain vaccine. Our study demonstrated that a single immunization with the r2segMP12 vaccine candidate at 105 plaque-forming units elicited a higher neutralizing antibody response than the rMP12 vaccine at the same vaccination titer without the need for a booster.

Estimation of the Minimal Rift Valley Fever Virus Protective Neutralizing Antibody Titer in Human Volunteers Immunized with MP-12 Vaccine Based on Protection in a Mouse Model of Disease.

The Rift Valley fever virus (RVFV) MP-12 vaccine is a promising human and veterinary vaccine. Although the vaccine elicited neutralizing antibody (nAb) in human volunteers, the minimal antibody titer that is needed to afford protection is unknown. Therefore, this study was conducted to determine the minimal nAb titer elicited by the RVFV MP-12 vaccine in human volunteers that protected mice against lethal RVFV challenge as a surrogate assessment of the protective efficacy of the vaccine. Among volunteers who were vaccinated with the MP-12 vaccine during a phase II trial, sera with antibody titers of 1:20 collected 5 years post-vaccination (PV), 1:40 titer collected 2 years PV, and 1:80 titer collected 1 year PV was passively transferred to groups of BALB/c mice. Blood samples were obtained 1 day after passive transfer to determine the RVFV neutralizing nAb titer before challenge with pathogenic RVFV (strain ZH501). Our results indicated that 1 day after passive transfer of the immune sera, an approximate 4-fold reduction in circulating nAb titers was detected in the mice. The presence of RVFV nAb titers in the range of 1:5 to 1:20 were generally protective (75-100% survival). These results suggested that circulating titers of 1:5 or higher offer a high degree of protection by MP-12-elicited antibody in human volunteers. Also, the findings highlighted the value of using the BALB/c mouse RVFV challenge model as a surrogate for evaluating the protective nAb responses elicited by MP-12 and possible use for evaluating the efficacy of other RVFV vaccine candidates.

Single-dose of a replication-competent adenovirus-vectored vaccine provides sterilizing protection against Rift Valley fever virus challenge

Rift Valley fever virus (RVFV) is one of the most important virulent pathogens causing severe disease in animals and humans. However, there is currently no approved vaccine to prevent RVFV infection in humans. The use of human adenovirus serotype 4 (Ad4) as a vector for an RVFV vaccine has not been reported. Here, we report the generation of a replication-competent recombinant Ad4 vector expressing codon-optimized forms of the RVFV glycoproteins Gn and Gc (named Ad4-GnGc). Intramuscular immunization with Ad4-GnGc elicited robust neutralizing antibodies against RVFV and cellular immune responses in mice. A single low-dose vaccination with Ad4-GnGc completely protected interferon-α/β receptor-deficient A129 mice from lethal RVFV infection. More importantly, Ad4-GnGc efficacy was not affected by pre-existing immunity to adenovirus serotype 5, which currently exists widely in populations. These results suggest that Ad4-GnGc is a promising vaccine candidate against RVFV.

Rift Valley fever virus 78kDa envelope protein attenuates virus replication in macrophage-derived cell lines and viral virulence in mice.

Rift Valley fever virus (RVFV) is a mosquito-borne bunyavirus with a wide host range including ruminants and humans. RVFV outbreaks have had devastating effects on public health and the livestock industry in African countries. However, there is no approved RVFV vaccine for human use in non-endemic countries and no FDA-approved antiviral drug for RVFV treatment. The RVFV 78kDa protein (P78), which is a membrane glycoprotein, plays a role in virus dissemination in the mosquito host, but its biological role in mammalian hosts remains unknown. We generated an attenuated RVFV MP-12 strain-derived P78-High virus and a virulent ZH501 strain-derived ZH501-P78-High virus, both of which expressed a higher level of P78 and carried higher levels of P78 in the virion compared to their parental viruses. We also generated another MP-12-derived mutant virus (P78-KO virus) that does not express P78. MP-12 and P78-KO virus replicated to similar levels in fibroblast cell lines and Huh7 cells, while P78-High virus replicated better than MP-12 in Vero E6 cells, fibroblast cell lines, and Huh7 cells. Notably, P78-High virus and P78-KO virus replicated less efficiently and more efficiently, respectively, than MP-12 in macrophage cell lines. ZH501-P78-High virus also replicated poorly in macrophage cell lines. Our data further suggest that inefficient binding of P78-High virus to the cells led to inefficient virus internalization, low virus infectivity and reduced virus replication in a macrophage cell line. P78-High virus and P78-KO virus showed lower and higher virulence than MP-12, respectively, in young mice. ZH501-P78-High virus also exhibited lower virulence than ZH501 in mice. These data suggest that high levels of P78 expression attenuate RVFV virulence by preventing efficient virus replication in macrophages. Genetic alteration leading to increased P78 expression may serve as a novel strategy for the attenuation of RVFV virulence and generation of safe RVFV vaccines.

A single vaccination with four-segmented Rift Valley fever virus prevents vertical transmission of the wild-type virus in pregnant ewes

Rift Valley fever virus (RVFV) is a mosquito-transmitted bunyavirus that causes severe outbreaks among wild and domesticated ruminants, of which sheep are the most susceptible. Outbreaks are characterised by high mortality rates among new-born lambs and abortion storms, in which all pregnant ewes in a flock may abort their foetuses. In endemic areas, Rift Valley fever (RVF) can be controlled by vaccination with either inactivated or live-attenuated vaccines. Inactivated vaccines are safe for animals during all physiological stages, including pregnancy. However, optimal efficacy of these vaccines depends on multiple vaccinations and yearly re-vaccination. Live-attenuated vaccines are generally highly efficacious after a single vaccination, but currently available live-attenuated vaccines may transmit to the ovine foetus, resulting in stillbirths, congenital malformations or abortion. We have previously reported the development of a novel live-attenuated RVFV vaccine, named RVFV-4s. This vaccine virus was created by splitting the M genome segment and deleting the major virulence determinant NSs, and was shown to be safe even for the most susceptible species, including pregnant ewes. The demonstrated efficacy and safety profile suggests that RVFV-4s holds promise for veterinary and human application. The RVFV-4s vaccine for veterinary application, here referred to as vRVFV-4s, was shown to provide complete protection after a single vaccination of lambs, goats and cattle. In this work, we evaluated the efficacy of the vRVFV-4s vaccine in pregnant ewes. Anticipating on the extremely high susceptibility of pregnant ewes for RVFV, both a single vaccination and double vaccination were evaluated in two independent experiments. The combined results suggest that a single vaccination with vRVFV-4s is sufficient to protect pregnant ewes and to prevent transmission to the ovine foetus.

Comparative thermo-stability of two Rift Valley fever virus vaccine candidate CL13T with a recombinant arMP-12ΔNSm21/384.

Rift Valley fever (RVF) is a zoonotic, viral disease, transmitted by mosquitoes, characterized by high mortality rates in young animals. RVF is an endemic and enzootic disease in the Arabian Peninsula and Africa, causing public health and economic instability. Therefore, it is important to develop vaccines to minimize outbreaks and combat the disease. We documented the stability of the thermo-stability of live attenuated RVF CL13T and recombinant arMP-12ΔNSm21/384 vaccine candidates at different temperatures, including these vaccine viruses in liquid and lyophilized form. The study revealed that both CL13T and recombinant arMP-12ΔNSm21/384 strains were stable for more than 18 months at 4°C. We show that at room temperatures (37°C and 45°C) the CL13T was less temperature sensitive than MP-12NSm-del in both lyophilized and liquid form. These findings are useful for the preparation of RVF vaccines that will avoid the need for a cold chain and therefore, will improve the application of the vaccines under field conditions.

Sensitivity of Lateral Flow technique for Evaluation of Inactivated Rift Valley Fever Virus Vaccine in Comparison with Serum Neutralization Test

Sensitivity of Lateral Flow technique for Evaluation of Inactivated Rift Valley Fever Virus Vaccine in Comparison with Serum Neutralization Test

Correlation Between Rift Valley Fever Virus (RVFV) Neutralizing Antibody Titers in Vaccinated Sheep and Effective Dose 50 (ED50) in Vaccinated Mice

Rift valley fever is an arthropod-born, multispecies zoonotic viral disease. Control of RVF disease depends mainly on vector control and vaccination of susceptible animals. The present work aims to detect the correlation between Rift Valley Fever Virus (RVFV) neutralizing antibody titers in vaccinated sheep using Serum Neutralization test as in vitro test and effective dose fifty in vaccinated mice as in vivo potency test and determine if they can be alternative to each other. In this work,17 inactivated RVFV vaccine batches were evaluated, applying SNT for serum samples of vaccinated sheep and ED50 in vaccinated mice. The two models of tests showed compatible results, where the same 14 vaccine batches showed satisfactory results [(SNT >1.5) and (ED50 less than 0.02)], while the other three batches revealed unsatisfactory results in both two tests. Statistical analysis of results using Wilcoxon’s test was (0.0001), indicating a significant correlation between the tests so it could be recommended to depend on SNT instead of mice inoculation in the evaluation of RVF vaccine to reduce the numbers of animals being used and to avoid the possible public health hazard.

Rift Valley fever virus vaccination induces long-lived, antigen-specific human T cell responses

Rift Valley fever virus (RVFV) is a zoonotic arbovirus of clinical significance in both livestock and humans. A formalin-inactivated virus preparation was initially developed for human use and tested in laboratory workers in the 1960s. Vaccination resulted in generation of neutralizing antibody titers in most recipients, but neutralization titers waned over time, necessitating frequent booster doses. In this study, T cell-based immune responses to the formalin-inactivated vaccine were examined in a cohort of seven individuals who received between 1 and 6 doses of the vaccine. RVFV-specific T cell responses were detectable up to 24 years post vaccination. Peripheral blood mononuclear cells from this cohort of individuals were used to map out the viral epitopes targeted by T cells in humans. These data provide tools for assessing human RVFV-specific T cell responses and are thus a valuable resource for future human RVFV vaccine efforts.

Chitosan and chitosan nanoparticles as adjuvant in local Rift Valley Fever inactivated vaccine.

The present study aimed to improve the potency of inactivated Rift Valley Fever Virus (RVFV) vaccine using chitosan (CS) or chitosan nanoparticles (CNP) as adjuvants. Chitosan nanoparticles were prepared by ionic gelation method. Rift Valley Fever Virus (RVFV) inactivated antigen was loaded on CS and CNP to form two vaccine formulations, RVFV-chitosan nanoparticles based vaccine (RVFV-CNP) and RVFV chitosan based vaccine (RVFV-CS). Five groups of mice were used in this study, each group was injected with one of the following: phosphate buffer saline (group1 G1), RVFV-CNP (G2), (RVF-CS) (G3), RVFV-Alum based vaccine (RVFV-Alum) (G4) and adjuvant free RVFV inactivated antigen (RVFV-Ag) (G5). The immunization was performed twice with 2 weeks interval. The results showed that, RVFV-CNP vaccine enhanced strongly the phagocytic activity of peritoneal macrophage (PM), neutralization antibodies titer against RVFV and IgG values against RVFV nucleoprotein than other vaccine formulations did. In addition, the RVFV-CNP and RVF-CS vaccines upregulate the gene expression of IL-2, IFN-γ (which promote cell mediated immunity) and IL-4 (which promote humeral immunity), while RVFV-Alum vaccine upregulate the gene expression of IL-4 only. These findings indicated that CS and CNP were comparable to the alum as adjuvant in efficacy but superior to it in inducing cell-mediated immune response and might be a candidate adjuvant for inactivated RVFV vaccine.

Genetically Modified Rabies Virus Vector-Based Rift Valley Fever Virus Vaccine is Safe and Induces Efficacious Immune Responses in Mice.

Rift Valley fever virus (RVFV), which causes Rift Valley fever (RVF), is a mosquito-borne zoonotic pathogen that causes serious morbidity and mortality in livestock and humans. RVF is a World Health Organization (WHO) priority disease and, together with rabies, is a major health burden in Africa. Here, we present the development and characterization of an inactivated recombinant RVFV and rabies virus (RABV) vaccine candidate (rSRV9-eGn). Immunization with rSRV9-eGn stimulated the production of RVFV-specific IgG antibodies and induced humoral and cellular immunity in mice but did not induce the production of neutralizing antibodies. IgG1 and IgG2a were the main isotypes observed by IgG subtype detection, and IgG3 antibodies were not detected. The ratios of IgG1/IgG2a > 1 indicated a Type 2 humoral immune response. An effective vaccine is intended to establish a long-lived population of memory T cells, and mice generated memory cells among the proliferating T cell population after immunization with rSRV9-eGn, with effector memory T cells (TEM) as the major population. Due to the lack of prophylactic treatment experiments, it is impossible to predict whether this vaccine can protect animals from RVFV infection with only high titres of anti-RVFV IgG antibodies and no neutralizing antibodies induced, and thus, protection confirmation needs further verification. However, this RVFV vaccine designed with RABV as the vector provides ideas for the development of vaccines that prevent RVFV and RABV infections.

In vitro and in vivo efficacy of a Rift Valley fever virus vaccine based on pseudovirus

ABSTRACT Rift Valley fever virus (RVFV), a recognized category A priority pathogen, causes large outbreaks of Rift Valley fever with some fatalities in humans in humans and huge economic losses in livestock. As wild-type RVFV must be handled in BSL-3 or BSL-4 laboratories, we constructed a high-titer vesicular stomatitis virus (VSV) pseudotype bearing RVFV envelope glycoproteins to detect neutralizing antibodies in vitro under BSL-2 conditions. The neutralizing properties of 39 amino acid mutant sites that have occurred naturally over time in the RVFV envelope glycoproteins were analyzed with their corresponding pseudoviral mutants separately. Compared with the results in the primary strain, the variants showed no statistically significant differences. We next established a Balb/c mouse pseudovirus infection model for detecting neutralizing antibodies against pseudovirus. Five immunizations with pseudoviral DNA protected the mice from infection with the pseudovirus. Bioluminescence imaging, which we used to evaluate viral dissemination and distribution in the mice, showed a good relationship between the neutralizing antibodies titers in vitro. These pseudovirus methods will allow for the safe determination of neutralizing antibodies in vivo and in vitro, and will assist with studies on vaccines and drugs against RVFV with the long term objective of Rift Valley fever prevention.

Evaluation of the Immunogenicity of a Mutagenized Rift Valley Fever MP-12 and a Recombinant Rift Valley Fever arMP12ΔNsm21/384 Vaccine Candidates in Indigenous Species of Cattle, Sheep, and Goats in Tanzania

Objective: Rift valley fever virus (RVFV) is the cause of devastating outbreaks among domestic ruminants and humans in most African countries and in the Arabian Peninsula. Current efforts to develop and evaluate an improved veterinary vaccine to prevent RVF disease among domestic ruminants includes a live attenuated RVFV MP-12 candidate and a recombinant vaccine referred to as arMP12ΔNsm21/384 that was derived from the RVF MP-12 vaccine candidate. The aim of this study was to evaluate these RVFV vaccine candidates in domestic ruminants in Tanzania. Methods: Six to nine months old goats (Capra aegagrus hircus), calves (Bos taurus indicus) and sheep (Ovis aries) were vaccinated subcutaneously with one ml each of 1 × 105 plaque forming units (PFU)/ml of the RVF MP-12 and/or the arMP-12ΔNSm21/384 candidate vaccines. The controls included six animals, two of each species which received 1 ml of Eagle's Minimum Essential Medium as a placebo. Blood samples were collected on day 2 before vaccination, and on the day (0) immediately before vaccination were tested for RVFV antibody to insure that only antibody negative animals were used in the vaccine trials. Samples collected on days, 3, 4, 5, 7 post-vaccination (PV) to determine the possibility of a vaccine induced viremia, and on days 4, 5, 7, 14, 21, 28, 35, 42 and 67 to determine the immune response of the vaccinated animals. Sera samples were tested for RVFV RNA by RT-PCR and for infectious virus in Vero E6 cells. The immune response was determined by testing sera samples for antibody using a commercial IDVERT ELISA kit (Montpellier-France) as well as a plaque reduction neutralization test (PRNT). Animals were observed daily for adverse effects and rectal temperature was recorded at the time of blood collection. Results: All vaccinated animals developed RVFV neutralizing antibodies with titers ranging from 1:10 in some animals as early as day 4 and 5 PV to as high as 1:160-1:2560 over the 67 day study period with no adverse effect observed in any of the animals. The antibody titers of goats to both MP-12 and arMP-12ΔNSm21/384 vaccines was significantly higher than the response observed for sheep and cattle. A viremia was not detected in any of the vaccinated and control animals. Conclusions: The findings of this study demonstrated that the RVFV MP-12 and arMP12ΔNsm21/384 vaccine candidates administered by the SC route elicited RVFV neutralizing antibodies in indigenous species of cattle, sheep, and goats in Tanzania, and therefore warrants further studies to assess the safety and protective efficacy of these vaccine candidates in domestic ruminants.

Preliminary evaluation of diagnostic accuracy and precision of a competitive ELISA for detection of antibodies to Rift Valley fever virus in cattle and sheep sera

Rift Valley fever virus (RVFV), a mosquito-borne, zoonotic pathogen, is endemic to sub-Saharan Africa and has spread beyond the continent to the Arabian Peninsula. The high likelihood of RVFV’s spread to other non-endemic countries spurs the need for development and implementation of rapid diagnostic tests and surveillance programs. In this preliminary evaluation, we assessed the diagnostic accuracy and precision of a recombinant RVFV nucleoprotein based competitive ELISA (cELISA) assay to detect RVFV antibodies compared to a plaque reduction neutralization test (PRNT80), using sera of cattle and sheep that were experimentally infected with either the MP-12 RVFV vaccine or a wild-type RVFV strain, as well as using known RVFV negative sera. With the manufacturer recommended 60% inhibition for the cELISA and a 1:40 titer for the PRNT80, the sensitivity and specificity of the cELISA assay was determined to be 95.1% (95% CI = 83.5–99.4%) and 91.8% (95% CI = 85.0–96.2%), respectively. Antibodies to RVFV were first detected at 5 days post inoculation (dpi) in both species’ sera. The prototype cELISA is an easy to implement, sensitive, specific, and safe test for the detection of antibodies to RVFV that can be used in surveillance programs.

Attenuation and efficacy of live-attenuated Rift Valley fever virus vaccine candidates in non-human primates.

Rift Valley fever virus (RVFV) is an important mosquito-borne veterinary and human pathogen that has caused large outbreaks of severe disease throughout Africa and the Arabian Peninsula. Currently, no licensed vaccine or therapeutics exists to treat this potentially deadly disease. The explosive nature of RVFV outbreaks and the severe consequences of its accidental or intentional introduction into RVFV-free areas provide the impetus for the development of novel vaccine candidates for use in both livestock and humans. Rationally designed vaccine candidates using reverse genetics have been used to develop deletion mutants of two known RVFV virulence factors, the NSs and NSm genes. These recombinant viruses were demonstrated to be protective and immunogenic in rats, mice, and sheep, without producing clinical illness in these animals. Here, we expand upon those findings and evaluate the single deletion mutant (ΔNSs rRVFV) and double deletion mutant (ΔNSs-ΔNSm rRVFV) vaccine candidates in the common marmoset (Callithrix jacchus), a non-human primate (NHP) model resembling severe human RVF disease. We demonstrate that both the ΔNSs and ΔNSs-ΔNSm rRVFV vaccine candidates were found to be safe and immunogenic in the current study. The vaccinated animals received a single dose of vaccine that led to the development of a robust antibody response. No vaccine-induced adverse reactions, signs of clinical illness or infectious virus were detected in the vaccinated marmosets. All vaccinated animals that were subsequently challenged with RVFV were protected against viremia and liver disease. In summary, our results provide the basis for further development of the ΔNSs and ΔNSs-ΔNSm rRVFV as safe and effective human RVFV vaccines for this significant public health threat.

Evaluation of Competitive ELISA For Detection of Antibodies to Rift Valley Fever Virus in Cattle and Sheep Sera

Rift Valley fever virus (RVFV) is a mosquito‐borne, zoonotic pathogen that causes mass abortions and deaths in ruminants. In humans, it can cause illnesses ranging from fever to occasionally fatal hemorrhagic fever, encephalitis and liver failure. RVFV is an enveloped, single‐stranded, negative‐sense tripartite RNA virus of the Phlebovirus genus (family: Phenuiviridae, order: Bunyavirales). It is categorized as a Select Agent by the CDC and USDA. Originally endemic to sub‐Saharan Africa only, it has recently spread beyond the continent to the Arabian Peninsula. Thus, the high likelihood of RVFV's spread to non‐endemic countries spurs the need for rapid diagnostics and surveillance tests. In this study, we assessed the efficacy of the recombinant RVFV nucleoprotein based competitive ELISA (cELISA) assay to detect RVFV antibodies in cattle and sheep sera. This cELISA is a new prototype packaged by Veterinary Medical Research & Development (VMRD). We used heat inactivated serum samples from ruminants (cattle=66, sheep=99) that were experimentally infected with either the MP‐12 RVFV vaccine or a virulent strain as well as known RVFV negative sera (cattle=330, sheep=179). We compared the cELISA assay with a plaque reduction neutralization test (PRNT80), the gold standard method for the detection of anti‐RVFV neutralizing antibodies. The recommended cut‐off value for the cELISA was 60%. Additionally, using experimental sera determined positive or negative by PRNT80 (at cut‐offs <1:10 and 1:40) and ROC analysis, we determined the optimal cut‐offs to be 46% and 68% respectively. With the cut‐off of 60% for cELISA and 1:40 titer for PRNT80, the sensitivity and specificity of the cELISA assay was determined to be 95.1% and 91.8% respectively. Antibodies to RVFV were first detected at 5 days post inoculation (dpi) in both sheep and cattle. Interestingly, at 5 dpi, the cELISA detected antibodies in 2/4 samples from RVFV inoculated cattle in comparison to 0/4 by PRNT80. We found the prototype cELISA to be an easy, sensitive, specific, and safe test for the detection of antibodies to RVFV in cattle and sheep. The prototype cELISA thus, could be used for early diagnosis and surveillance that will help in diminish the disease burden.Support or Funding InformationThis project was funded by the Science and Technology Directorate of the U.S. Department of Homeland Security under Award Number HSHQDC‐13‐J‐00278 to the Institute for Infectious Animal Diseases within the Texas A&M University System, the USDA, Agricultural Research Service Project # 5430‐050‐009‐00D, and Department of Homeland Security Center of Excellence for Emerging and Zoonotic Animal Diseases (CEEZAD), Grant No. 2010‐ST061‐AG0001. The views and conclusions expressed in this publication are those of authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of the U.S. Department of Homeland Security or the USDA.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.