Production and Characterization of Monoclonal Antibodies Against N Protein of Rift Valley Fever Virus.

The DNA fragment encoding predicted main antigenic region, aa 14-245 on N protein of Rift Valley virus (RVFV) was cloned into the vector pET-28a (+) and p3xFLAG-CMV-10. The recombinant pET-28a-N1 protein was expressed in Escherichia coli BL21 (DE3) with 1 mM isopropyl-b-thio-galactopyranoside at 37°C for 5 hours, and purified by protein purifier. Three monoclonal antibodies (mAbs) named 3A5, 3A6, and 3A7 against N protein were obtained by fusing mouse myeloma cell line SP2/0 with spleen lymphocytes from pET-28a-N1 protein-immunized mice. Finally, the mAbs were characterized by enzyme-linked immunosorbent assays, indirect immunofluorescent assays, and Western blot. The results show that all the mAbs possess high specificity and react with both prokaryotic and eukaryotic N protein, which could provide important materials for the research on the function of N protein and the diagnostic methods of RVFV.

The challenging management of Rift Valley Fever in humans: literature review of the clinical disease and algorithm proposal.

Rift Valley Fever (RVF) is an emerging zoonotic arbovirus with a complex cycle of transmission that makes difficult the prediction of its expansion. Recent outbreaks outside Africa have led to rediscover the human disease but it remains poorly known. The wide spectrum of acute and delayed manifestations with potential unfavorable outcome much complicate the management of suspected cases and prediction of morbidity and mortality during an outbreak. We reviewed literature data on bio-clinical characteristics and treatments of RVF human illness. We identified gaps in the field and provided a practical algorithm to assist clinicians in the cases assessment, determination of setting of care and prolonged follow-up.

Human immune cell engraftment does not alter development of severe acute Rift Valley fever in mice.

Rift Valley fever (RVF) in humans is usually mild, but, in a subset of cases, can progress to severe hepatic and neurological disease. Rodent models of RVF generally develop acute severe clinical disease. Here, we inoculated humanized NSG-SGM3 mice with Rift Valley fever virus (RVFV) to investigate whether the presence of human immune cells in mice would alter the progression of RVFV infection to more closely model human disease. Despite increased human cytokine expression, including responses mirroring those seen in human disease, and decreased hepatic viral RNA levels at terminal euthanasia, both high- and low-dose RVFV inoculation resulted in lethal disease in all mice with comparable time-to-death as unengrafted mice.

Rift Valley Fever.

Rift Valley fever (RVF) is a severe veterinary disease of livestock that also causes moderate to severe illness in people. The life cycle of RVF is complex and involves mosquitoes, livestock, people, and the environment. RVF virus is transmitted from either mosquitoes or farm animals to humans, but is generally not transmitted from person to person. People can develop different diseases after infection, including febrile illness, ocular disease, hemorrhagic fever, or encephalitis. There is a significant risk for emergence of RVF into new locations, which would affect human health and livestock industries.

Hemorrhagic fever of bunyavirus etiology: disease models and progress towards new therapies.

A growing number of bunyaviruses are known to cause viral hemorrhagic fever (VHF), a severe febrile illness which can progress to hypovolemic shock and multi-organ failure and is characterized by hematologic abnormalities and vascular leak. At present, there are no approved vaccines or antiviral therapies to effectively prevent or treat VHF caused by pathogenic bunyaviruses. Advances in the modeling of bunyaviral infections have facilitated efforts towards the development of novel post-exposure prophylactic and therapeutic countermeasures, several of which may some day be approved for human use. Here, we review recent progress in animal models of severe bunyaviral infections essential to this mission, as well as promising antivirals and biologicals that are at various stages of the development process.

Development of novel mechanisms for housing, handling, and remote monitoring of common marmosets at animal biosafety level 3.

The use of common marmosets as an alternative non-human primate model for infectious disease research using BSL-3 viruses such as Rift Valley fever virus (RVFV) presents unique challenges with respect to housing, handling, and safety. Subject matter experts from veterinary care, animal husbandry, biosafety, engineering, and research were consulted to design a pilot experiment using marmosets infected with RVFV. This paper reviews the caging, handling, and safety-related adaptations and modifications that were required to humanely utilize marmosets as a model for high-hazard BSL-3 viral diseases.

Aerosol exposure to Rift Valley fever virus causes earlier and more severe neuropathology in the murine model, which has important implications for therapeutic development.

Rift Valley fever virus (RVFV) is an important mosquito-borne veterinary and human pathogen that can cause severe disease including acute-onset hepatitis, delayed-onset encephalitis, retinitis and blindness, or a hemorrhagic syndrome. Currently, no licensed vaccine or therapeutics exist to treat this potentially deadly disease. Detailed studies describing the pathogenesis of RVFV following aerosol exposure have not been completed and candidate therapeutics have not been evaluated following an aerosol exposure. These studies are important because while mosquito transmission is the primary means for human infection, it can also be transmitted by aerosol or through mucosal contact. Therefore, we directly compared the pathogenesis of RVFV following aerosol exposure to a subcutaneous (SC) exposure in the murine model by analyzing survival, clinical observations, blood chemistry, hematology, immunohistochemistry, and virus titration of tissues. Additionally, we evaluated the effectiveness of the nucleoside analog ribavirin administered prophylactically to treat mice exposed by aerosol and SC. The route of exposure did not significantly affect the survival, chemistry or hematology results of the mice. Acute hepatitis occurred despite the route of exposure. However, the development of neuropathology occurred much earlier and was more severe in mice exposed by aerosol compared to SC exposed mice. Mice treated with ribavirin and exposed SC were partially protected, whereas treated mice exposed by aerosol were not protected. Early and aggressive viral invasion of brain tissues following aerosol exposure likely played an important role in ribavirin's failure to prevent mortality among these animals. Our results highlight the need for more candidate antivirals to treat RVFV infection, especially in the case of a potential aerosol exposure. Additionally, our study provides an account of the key pathogenetic differences in RVF disease following two potential exposure routes and provides important insights into the development and evaluation of potential vaccines and therapeutics to treat RVFV infection.

Rift Valley fever: the Nigerian story.

Rift Valley fever (RVF) is an arthropod-borne zoonotic disease of livestock. It is characterised by fever, salivation, abdominal pain, diarrhoea, mucopurulent to bloody nasal discharge, abortion, rapid decrease in milk production and death in animals. Infected humans experience an influenza-like illness that is characterised by fever, malaise, headaches, nausea and epigastric pain followed by recovery, although mortality can occur. RVF was thought to be a disease of sub-Saharan Africa but with the outbreaks in Egypt and the Arabian Peninsula, it may be extending its range further afield. Virological and serological evidence indicates that the virus exists in Nigeria and, with the warning signal sent by international organisations to countries in Africa about an impending outbreak, co-ordinated research between veterinarians and physicians in Nigeria is advocated.

[Rift Valley fever virus: evolution in progress]. / Le virus de la Fièvre de la Vallée du Rift: évolution en cours.

Several viruses now circulating in tropical zones around the globe are potential threats for ever-increasing human populations even in temperate zones that have long remained unaffected. The mechanisms underlying transport and transmission, which can be enhanced by human activity, can be even stronger in zones where factors needed to support development of these viruses, i.e., hosts, reservoirs and vectors, are already present. This possibility has been illustrated by dengue virus, and now by the rapid spread of the Chikungunya virus on Reunion Island in 2005 and then in Italy in 2007. The spreading of Chikungunya virus despite its mild reputation had a major unexpected impact. It showed that the evolution of the virus, whether a cause or consequence of observed events, could be determinant. The risk of extension of more pathogenic viruses due to similar mechanisms must be considered as a possibility. In this regard the Rift Valley fever virus, that already involves a large area and has a major reservoir, is one of the viruses that deserves close surveillance.

Application of knowledge-driven spatial modelling approaches and uncertainty management to a study of Rift Valley fever in Africa.

BACKGROUND: There are few studies that have investigated uncertainties surrounding the scientific community's knowledge of the geographical distribution of major animal diseases. This is particularly relevant to Rift Valley fever (RVF), a zoonotic disease causing destructive outbreaks in livestock and man, as the geographical range of the disease is widening to involve previously unaffected regions. In the current study we investigate the application of methods developed in the decision sciences: multiple criteria decision making using weighted linear combination and ordered weighted averages, and Dempster-Shafer theory, implemented within the geographical information system IDRISI, to obtain a greater understanding of uncertainty related to the geographical distribution of RVF. The focus is on presenting alternate methods where extensive field data are not available and traditional, model-based approaches to disease mapping are impossible to conduct. RESULTS: Using a compensatory multiple criteria decision making model based on weighted linear combination, most of sub-Saharan Africa was suitable for endemic circulation of RVF. In contrast, areas where rivers and lakes traversed semi-arid regions, such as those bordering the Sahara, were highly suitable for RVF epidemics and wet, tropical areas of central Africa had low suitability. Using a moderately non-compensatory model based on ordered weighted averages, the areas considered suitable for endemic and epidemic RVF were more restricted. Varying the relative weights of the different factors in the models did not affect suitability estimates to a large degree, but variations in model structure had a large impact on our suitability estimates. Our Dempster-Shafer analysis supported the belief that a range of semi-arid areas were suitable for RVF epidemics and the plausibility that many other areas of the continent were suitable. Areas where high levels of uncertainty were highlighted included the Ethiopian Highlands, southwest Kenya and parts of West Africa. CONCLUSION: We have demonstrated the potential of methods developed in the decision sciences to improve our understanding of uncertainties surrounding the geographical distribution of animal diseases, particularly where information is sparse, and encourage wider application of the decision science methodology in the field of animal health.

Rift Valley fever virus.

Rift Valley fever is considered to be one of the most important viral zoonoses in Africa. In 2000, the Rift valley fever virus spread to the Arabian Peninsula and caused two simultaneous outbreaks in Yemen and Saudi Arabia. It is transmitted to ruminants and to humans by mosquitoes. The viral agent is an arbovirus, which belongs to the Phlebovirus genus in the Bunyaviridae family. This family of viruses comprises more than 300 members grouped into five genera: Orthobunyavirus, Phlebovirus, Hantavirus, Nairovirus, and Tospovirus. Several members of the Bunyaviridae family are responsible for fatal hemorrhagic fevers: Rift Valley fever virus (Phlebovirus), Crimean-Congo hemorrhagic fever virus (Nairovirus), Hantaan, Sin Nombre and related viruses (Hantavirus), and recently Garissa, now identified as Ngari virus (Orthobunyavirus). Here are reviewed recent advances in Rift Valley fever virus, its epidemiology, molecular biology and focus on recent data on the interactions between viral and cellular proteins, which help to understand the molecular mechanisms utilized by the virus to circumvent the host cellular response.

Potential role of immunomodulators for treatment of phlebovirus infections of animals.

Rift Valley fever (RVFV) is a major phlebovirus-induced epizootic disease of domestic animals (primarily cattle and sheep) in Africa. No therapies for the disease are known. A related phlebovirus, Punta Toro virus (PTV), has been adapted to induce an RVFV-like disease in C57BL/6 mice. This PTV infection has been used as a model for RVFV because it is reasonably safe and does not require high-level biologic containment. The infection model has been used to study the potential role of immunomodulating substances as therapies. A spectrum of immunomodulators has been studied; those immunomodulators most capable of preventing death and other disease manifestations are ampligen, bropirimine, poly (ICLC), AM-3, P-136, and 7-thia-8-oxoguanosine. An immunologic parameter common to all these substances has been their ability to induce interferon. Timing studies have indicated that these active substances may be administered therapeutically as well as prophylactically to inhibit markedly the progress of the disease. Further work is needed in the development of these materials for use in treating viral infections in domestic animals. As a next step, studies need to be run to compare the immunologic profiles induced by each substance in domestic animals and in mice.

Prevention of Rift Valley fever in rhesus monkeys with interferon-alpha.

Prophylactic and therapeutic efficacy of recombinant leukocyte A interferon (rIFN-alpha A) and Sendai virus-induced human leukocyte interferon (HuIFN-alpha) administered intramuscularly to Rift Valley fever virus (RVFV)-infected rhesus monkeys was studied. Clinical, virologic, immunologic, and hemostatic parameters were monitored. Five daily inoculations of 5 X 10(5) units of either interferon product per kilogram of body weight, initiated 24 hours before or 6 hours after RVFV infection, prevented or greatly suppressed viremia. No clinical signs of disease or laboratory evidence of impaired hemostasis was observed. Serum neutralizing antibody to RVFV was detected within 6 days of virus inoculation. Prophylactic administration of 5 X 10(4) or 5 X 10(3) units of rIFN-alpha A per kilogram also limited viremia, hepatocellular damage, and hemostatic derangement. Untreated, RVFV-infected, control monkeys developed high-titered viremia, clinical disease, and impaired hemostasis. These data suggest that rIFN-alpha A and HuIFN-alpha are effective in protecting RVFV-infected rhesus monkeys from viremia and hepatocellular damage and may be beneficial in human RVF infection.