CHARACTERIZATION OF NIPAH VIRUS PATHOGENICITY IN VITRO AND IN VIVO AND PROTECTION FROM DISEASE USING A SINGLE DOSE RECOMBINANT VACCINE

Nipah virus is a zoonotic pathogen that infects a wide species range including humans. It was first discovered in Malaysia in 1998 during a large outbreak, but since has spread to Bangladesh and India causing almost yearly outbreaks in the region since 2001. The distinct geographic locations have led to two genetically varied strains. Infection of humans by Nipah virus leads to respiratory distress and acute encephalitis. Pathology caused by the virus is characterized by vasculitis, necrosis, and edema of small vessels of the lung and brain primarily. This work investigates differences of pathology and clinical signs in the hamster model between strains, aiming to explain differences seen in epidemiology reports. We also characterize infection of endothelial and smooth muscle cells, which make up the vasculature, and how they react to infection. After better understanding the pathology in vivo and in vitro, we developed and efficacy tested a single-dose Vesicular stomatitis virus based vaccine. Data from this work demonstrates that although the Bangladesh strain is delayed 2 days compared to the Malaysian strain they cause similar pathology in hamsters. We also show that Nipah virus replicated in smooth muscle cells but does not cause adverse effects. Finally this study presents a vaccine that is protective against Nipah virus pathology. Overall this work allows for future studies using the Bangladesh strain, better defines infection of primary vascular cells, and proposes a possible vaccine candidate for outbreak use

Nipah Virus Efficiently Replicates in Human Smooth Muscle Cells without Cytopathic Effect

Nipah virus (NiV) is a highly pathogenic zoonotic virus with a broad species tropism, originating in pteropid bats. Human outbreaks of NiV disease occur almost annually, often with high case-fatality rates. The specific events that lead to pathogenesis are not well defined, but the disease has both respiratory and encephalitic components, with relapsing encephalitis occurring in some cases more than a year after initial infection. Several cell types are targets of NiV, dictated by the expression of the ephrin-B2/3 ligand on the cell’s outer membrane, which interact with the NiV surface proteins. Vascular endothelial cells (ECs) are major targets of infection. Cytopathic effects (CPE), characterized by syncytia formation and cell death, and an ensuing vasculitis, are a major feature of the disease. Smooth muscle cells (SMCs) of the tunica media that line small blood vessels are infected in humans and animal models of NiV disease, although pathology or histologic changes associated with antigen-positive SMCs have not been reported. To gain an understanding of the possible contributions that SMCs might have in the development of NiV disease, we investigated the susceptibility and potential cytopathogenic changes of human SMCs to NiV infection in vitro. SMCs were permissive for NiV infection and resulted in high titers and prolonged NiV production, despite a lack of cytopathogenicity, and in the absence of detectable ephrin-B2/3. These results indicate that SMC might be important contributors to disease by producing progeny NiV during an infection, without suffering cytopathogenic consequences.Peer Reviewe

Designing for Sustainability:Lessons Learned from Four Industrial Projects

Scientific research addressing the relation between software and sustainability is slowly maturing in two focus areas, related to `sustainable software' and `software for sustainability'. The first is better understood and may include research foci like energy-efficient software and software maintainability. It most-frequently covers `technical' concerns. The second, `software for sustainability', is much broader in both scope and potential impact, as it entails how software can contribute to sustainability goals in any sector or application domain. Next to the technical concerns, it may also cover economic, social, and environmental sustainability. Differently from researchers, practitioners are often not aware or well-trained in all four types of software sustainability concerns. To address this need, in previous work we have defined the Sustainability-Quality Assessment Framework (SAF) and assessed its viability via the analysis of a series of software projects. Nevertheless, it was never used by practitioners themselves, hence triggering the question: What can we learn from the use of SAF in practice? To answer this question, we report the results of practitioners applying the SAF to four industrial cases. The results show that the SAF helps practitioners in (1) creating a sustainability mindset in their practices, (2) uncovering the relevant sustainability-quality concerns for the software project at hand, and (3) reasoning about the inter-dependencies and trade-os of such concerns as well as the related short- and long-term implications. Next to improvements for the SAF, the main lesson for us as researchers is the missing explicit link between the SAF and the (technical) architecture design

Syrian Hamster as an Animal Model for the Study on Infectious Diseases

Infectious diseases still remain one of the biggest challenges for human health. In order to gain a better understanding of the pathogenesis of infectious diseases and develop effective diagnostic tools, therapeutic agents, and preventive vaccines, a suitable animal model which can represent the characteristics of infectious is required. The Syrian hamster immune responses to infectious pathogens are similar to humans and as such, this model is advantageous for studying pathogenesis of infection including post-bacterial, viral and parasitic pathogens, along with assessing the efficacy and interactions of medications and vaccines for those pathogens. This review summarizes the current status of Syrian hamster models and their use for understanding the underlying mechanisms of pathogen infection, in addition to their use as a drug discovery platform and provides a strong rationale for the selection of Syrian hamster as animal models in biomedical research. The challenges of using Syrian hamster as an alternative animal model for the research of infectious diseases are also addressed. Keywords: infectious diseases, Syrian hamster, drug discovery, infection mechanism, biomedical researc

Crystal Structure of the Hendra Virus Attachment G Glycoprotein Bound to a Potent Cross-Reactive Neutralizing Human Monoclonal Antibody

The henipaviruses, represented by Hendra (HeV) and Nipah (NiV) viruses are highly pathogenic zoonotic paramyxoviruses with uniquely broad host tropisms responsible for repeated outbreaks in Australia, Southeast Asia, India and Bangladesh. The high morbidity and mortality rates associated with infection and lack of licensed antiviral therapies make the henipaviruses a potential biological threat to humans and livestock. Henipavirus entry is initiated by the attachment of the G envelope glycoprotein to host cell membrane receptors. Previously, henipavirus-neutralizing human monoclonal antibodies (hmAb) have been isolated using the HeV-G glycoprotein and a human naïve antibody library. One cross-reactive and receptor-blocking hmAb (m102.4) was recently demonstrated to be an effective post-exposure therapy in two animal models of NiV and HeV infection, has been used in several people on a compassionate use basis, and is currently in development for use in humans. Here, we report the crystal structure of the complex of HeV-G with m102.3, an m102.4 derivative, and describe NiV and HeV escape mutants. This structure provides detailed insight into the mechanism of HeV and NiV neutralization by m102.4, and serves as a blueprint for further optimization of m102.4 as a therapeutic agent and for the development of entry inhibitors and vaccines

A Human Lung Xenograft Mouse Model of Nipah Virus Infection

Nipah virus (NiV) is a member of the genus Henipavirus (family Paramyxoviridae) that causes severe and often lethal respiratory illness and encephalitis in humans with high mortality rates (up to 92%). NiV can cause Acute Lung Injury (ALI) in humans, and human-to-human transmission has been observed in recent outbreaks of NiV. While the exact route of transmission to humans is not known, we have previously shown that NiV can efficiently infect human respiratory epithelial cells. The molecu