scholarly journals A Novel System for Rapid and Cost-Effective Production of Detection and Diagnostic Reagents of West Nile Virus in Plants

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Junyun He ◽  
Huafang Lai ◽  
Christopher Brock ◽  
Qiang Chen
Keyword(s):  
West Nile Virus ◽  
Posttranslational Modifications ◽  
Protein Production ◽  
Low Cost ◽  
Expression System ◽  
Cost Effective ◽  
West Nile ◽  
Economical System ◽  
High Scalability ◽  
Detection And Diagnosis

The threat of West Nile virus (WNV) epidemics necessitates the development of a technology platform that can produce reagents to support detection and diagnosis rapidly and inexpensively. A plant expression system is attractive for protein production due to its low-cost and high-scalability nature and its ability to make appropriate posttranslational modifications. Here, we investigated the feasibility of using plants to produce two WNV detection and diagnostic reagents to address the current cost and scalability issues. We demonstrated that WNV DIII antigen and E16 monoclonal antibody are rapidly produced at high levels in two plant species and are easily purified. Furthermore, they are effective in identifying WNV and in detecting human IgM response to WNV infection. E16 mAb does not cross-react with other flaviviruses, therefore, is valuable for improving diagnostic accuracy. This study provides a proof of principle for using plants as a robust and economical system to produce diagnostic reagents for arboviruses.

scholarly journals A Plant-Produced Antigen Elicits Potent Immune Responses against West Nile Virus in Mice

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Junyun He ◽  
Li Peng ◽  
Huafang Lai ◽  
Jonathan Hurtado ◽  
Jake Stahnke ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Immune Responses ◽  
Viral Vector ◽  
Specific Binding ◽  
Low Cost ◽  
Expression System ◽  
Conformational Epitope ◽  
West Nile ◽  
Domain Iii ◽  
Subcutaneous Immunization

We described the rapid production of the domain III (DIII) of the envelope (E) protein in plants as a vaccine candidate for West Nile Virus (WNV). Using various combinations of vector modules of a deconstructed viral vector expression system, DIII was produced in three subcellular compartments in leaves ofNicotiana benthamianaby transient expression. DIII expressed at much higher levels when targeted to the endoplasmic reticulum (ER) than that targeted to the chloroplast or the cytosol, with accumulation level up to 73 μg DIII per gram of leaf fresh weight within 4 days after infiltration. Plant ER-derived DIII was soluble and readily purified to > 95% homogeneity without the time-consuming process of denaturing and refolding. Further analysis revealed that plant-produced DIII was processed properly and demonstrated specific binding to an anti-DIII monoclonal antibody that recognizes a conformational epitope. Furthermore, subcutaneous immunization of mice with 5 and 25 μg of purified DIII elicited a potent systemic response. This study provided the proof of principle for rapidly producing immunogenic vaccine candidates against WNV in plants with low cost and scalability.

scholarly journals Plant-Produced Antigen Displaying Virus-Like Particles Evokes Potent Antibody Responses against West Nile Virus in Mice

Vaccines ◽  
2021 ◽  
Vol 9 (1) ◽  
pp. 60
Author(s):  
Junyun He ◽  
Huafang Lai ◽  
Adrian Esqueda ◽  
Qiang Chen
Keyword(s):  
West Nile Virus ◽  
Fusion Protein ◽  
Nicotiana Benthamiana ◽  
Specific Binding ◽  
Low Cost ◽  
Gradient Centrifugation ◽  
Conformational Epitope ◽  
Protein Domain ◽  
Core Antigen ◽  
West Nile

In this study, we developed a hepatitis B core antigen (HBcAg)-based virus-like particle (VLP) that displays the West Nile virus (WNV) Envelope protein domain III (wDIII) as a vaccine candidate for WNV. The HBcAg-wDIII fusion protein was quickly produced in Nicotiana benthamiana plants and reached a high expression level of approximately 1.2 mg of fusion protein per gram of leaf fresh weight within six days post gene infiltration. Electron microscopy and gradient centrifugation analysis indicated that the introduction of wDIII did not interfere with VLP formation and HBcAg-wDIII successfully assembled into VLPs. HBcAg-wDIII VLPs can be easily purified in large quantities from Nicotiana benthamiana leaves to >95% homogeneity. Further analysis revealed that the wDIII was displayed properly and demonstrated specific binding to an anti-wDIII monoclonal antibody that recognizes a conformational epitope of wDIII. Notably, HBcAg-wDIII VLPs were shown to be highly immunogenic and elicited potent humoral responses in mice with antigen-specific IgG titers equivalent to that of protective wDIII antigens in previous studies. Thus, our wDIII-based VLP vaccine offers an attractive option for developing effective, safe, and low-cost vaccines against WNV.

scholarly journals Twenty Years of Progress Toward West Nile Virus Vaccine Development

Viruses ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 823 ◽  
Author(s):  
Jaclyn A. Kaiser ◽  
Alan D.T. Barrett
Keyword(s):  
Clinical Trials ◽  
Single Dose ◽  
West Nile Virus ◽  
Vaccine Development ◽  
Cost Effective ◽  
Cumulative Number ◽  
West Nile ◽  
Transmitted Infection ◽  
Large Outbreak ◽  
Human Vaccine

Although West Nile virus (WNV) has been a prominent mosquito-transmitted infection in North America for twenty years, no human vaccine has been licensed. With a cumulative number of 24,714 neurological disease cases and 2314 deaths in the U.S. since 1999, plus a large outbreak in Europe in 2018 involving over 2000 human cases in 15 countries, a vaccine is essential to prevent continued morbidity, mortality, and economic burden. Currently, four veterinary vaccines are licensed, and six vaccines have progressed into clinical trials in humans. All four veterinary vaccines require multiple primary doses and annual boosters, but for a human vaccine to be protective and cost effective in the most vulnerable older age population, it is ideal that the vaccine be strongly immunogenic with only a single dose and without subsequent annual boosters. Of six human vaccine candidates, the two live, attenuated vaccines were the only ones that elicited strong immunity after a single dose. As none of these candidates have yet progressed beyond phase II clinical trials, development of new candidate vaccines and improvement of vaccination strategies remains an important area of research.

scholarly journals Immunogenicity and Protective Efficacy of a Recombinant Subunit West Nile Virus Vaccine in Rhesus Monkeys

2009 ◽  
Vol 16 (9) ◽  
pp. 1332-1337 ◽  
Author(s):  
Michael M. Lieberman ◽  
Vivek R. Nerurkar ◽  
Haiyan Luo ◽  
Bruce Cropp ◽  
Ricardo Carrion ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Cellular Immunity ◽  
Cytokine Production ◽  
Protective Efficacy ◽  
Expression System ◽  
High Yield ◽  
Separate Experiment ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
West Nile

ABSTRACT The immunogenicity and protective efficacy of a recombinant subunit West Nile virus (WNV) vaccine was evaluated in rhesus macaques (Macaca mulatta). The vaccine consisted of a recombinant envelope (E) protein truncated at the C-terminal end, resulting in a polypeptide containing 80% of the N-terminal amino acids of the native WNV protein (WN-80E), mixed with an adjuvant (GPI-0100). WN-80E was produced in a Drosophila melanogaster expression system with high yield and purified by immunoaffinity chromatography using a monoclonal antibody specific for flavivirus E proteins. Groups of monkeys were vaccinated with formulations containing 1 or 25 μg of WN-80E antigen, and both humoral and cellular immunity were assessed after vaccination. The results demonstrated potent antibody responses to vaccination, as determined by both enzyme-linked immunosorbent assay and virus-neutralizing antibody assays. All vaccinated animals responded favorably, and there was little difference in response between animals immunized with 1 or 25 μg of WN-80E. Cellular immunity was determined by lymphocyte proliferation and cytokine production assays using peripheral blood mononuclear cells from vaccinated animals stimulated in vitro with WN-80E. Cell-mediated immune responses varied from animal to animal within each group. About half of the animals responded with lymphoproliferation, cytokine production, or both. Again, there was little difference in response between animals immunized with a 1- or 25-μg dose of WN-80E in the vaccine formulations. In a separate experiment, groups of monkeys were immunized with the WN-80E/GPI-0100 vaccine or an adjuvant-only control formulation. Animals were then challenged by inoculation of wild-type WNV, and the level of viremia in each animal was monitored daily for 10 days. The results showed that whereas all animals in the control group had detectable viremia for at least 3 days after challenge, all of the vaccinated animals were negative on all days after challenge. Thus, the WN-80E vaccine was 100% efficacious in protecting monkeys against infection with WNV.

scholarly journals Novel infectious diseases in europe

2017 ◽  
Vol 70 (11-12) ◽  
pp. 385-390
Author(s):  
Dajana Lendak ◽  
Tomislav Preveden ◽  
Nadica Kovacevic ◽  
Slavica Tomic ◽  
Maja Ruzic ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Infectious Diseases ◽  
Zika Virus ◽  
Ebola Virus ◽  
Cost Effective ◽  
Hepatitis E ◽  
West Nile ◽  
Human Contact ◽  
Fast Spreading ◽  
Virus Zika

Introduction. The end of 20th and beginning of 21st century is marked by the discovery of new, supercontagious and fast spreading viral diseases. Since 1967, more than 40 new agents have been identified, including human immunodeficiency virus, Ebola, Marburg fever, severe acute respiratory syndrome, hepatitis C, hepatitis E viruses and Zika virus. Modern lifestyle, availability and speed of air traffic, migrations, as well as climate changes, enable faster spreading of infectious diseases from the regions that were hardly reachable. We selected a few diseases that raised the greatest attention among experts and public in general. Ebola. Ebola virus raises anxiety due to high mortality and fast spreading by using inter-human contact. Zika virus. Zika virus, that most often causes mild symptoms, is potentially responsible for microcephaly in neonates. Dengue. Dengue virus is an ?old story?, but in last decades incidence has multiplied by 30. West Nile virus. Although discovered in 1937, West Nile virus has been found exclusively in rural parts of Africa, while nowadays it represents one of the most important etiological factors of viral meningo-encephalitis all over the world. Hepatitis E. Today it is well-known that hepatitis E virus can cause not only acute viral hepatitis but also potentially blood-transmitted chronic hepatitis in immunocompromised, as well as some neurological disorders. Conclusion. One of the scientific challenges in the future will certainly be the discovery of available and cost-effective diagnostic tests, as well as efficient and safe vaccines for these diseases. Up to now, efficient prophylaxis is available only for Denga virus.

scholarly journals Mosquito excreta reveals circulation of West Nile virus and its underlying ecosystem

2021 ◽  
Author(s):  
Grégory L’Ambert ◽  
Mathieu Gendrot ◽  
Sébastien Briolant ◽  
Agnès Nguyen ◽  
Sylvain Pages ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Febrile Illness ◽  
Cost Effective ◽  
Community Diversity ◽  
Entomological Surveillance ◽  
West Nile ◽  
Non Invasive ◽  
Virus Surveillance ◽  
Vertebrate Hosts ◽  
Asymptomatic Infections

AbstractEmerging and endemic mosquito-borne viruses can be difficult to detect and monitor because they often cause asymptomatic infections in human or vertebrate animals or cause nonspecific febrile illness with a short recovery waiting period. Cases’ detection in vertebrate hosts can be complemented by entomological surveillance, but this method is not adapted to low infection rates in mosquito populations that typically occur in low or non-endemic areas. We identified West Nile Virus circulation in Camargue, a wetland area in South of France, using a cost effective innovative xenomonitoring method based on the molecular detection of virus in excreta from trapped mosquitoes. We also succeeded at identifying the mosquito community diversity dynamic on several sampling sites, together with the vertebrate hosts on which they fed prior to be captured using amplicon-based metagenomic on mosquito excreta without processing any mosquito. Mosquito excreta-based virus surveillance can be considered as a cost-effective and non-invasive strategy that offers the additional asset to reveal the ecological network underlying arbovirus circulation.

scholarly journals The Role of Birds of Prey in West Nile Virus Epidemiology

Vaccines ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 550 ◽  
Author(s):  
Beatriz Vidaña ◽  
Núria Busquets ◽  
Sebastian Napp ◽  
Elisa Pérez-Ramírez ◽  
Miguel Ángel Jiménez-Clavero ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Current Knowledge ◽  
Critical Role ◽  
Cost Effective ◽  
Birds Of Prey ◽  
West Nile ◽  
Transmission Cycle ◽  
Mediterranean Countries ◽  
Dead End ◽  
Lineage 2

Reported human cases of West Nile virus (WNV) in Europe increased dramatically in 2018. Lineage 1 strains had been circulating in Euro-Mediterranean countries since the early 1990s. The subsequent introduction of WNV lineage 2 has been responsible for the remarkable upsurge of European WNV outbreaks since 2004, including the dramatic increase in human cases observed since 2018. The virus exists in a natural cycle between mosquitoes and wild birds, with humans and horses acting as dead-end hosts. As the key vertebrate hosts in the transmission cycle of WNV, avian species have been the focus of surveillance across many countries. Raptors appear particularly susceptible to WNV infection, resulting in higher prevalence, and in some cases exhibiting neurological signs that lead to the death of the animal. In addition, birds of prey are known to play an important role as WNV reservoir and potentially amplifying hosts of infection. Importantly, raptor higher susceptibility/prevalence may indicate infection through predation of infected prey. Consequently, they are considered important target species when designing cost-effective surveillance for monitoring both seasonal WNV circulation in endemic countries and its emergence into new areas, where migrating raptors may play a critical role in virus introduction. This review summarizes the different aspects of the current knowledge of WNV infection in birds of prey and evaluates their role in the evolution of the epizootic that is spreading throughout Europe.

scholarly journals Plant Molecular Farming: A Viable Platform for Recombinant Biopharmaceutical Production

Plants ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 842 ◽  
Author(s):  
Balamurugan Shanmugaraj ◽  
Christine Joy I. Bulaon ◽  
Waranyoo Phoolcharoen
Keyword(s):  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Molecular Farming ◽  
Expression System ◽  
Cost Effective ◽  
Biologically Active ◽  
Native Proteins ◽  
Plant Molecular Farming ◽  
Major Factors

The demand for recombinant proteins in terms of quality, quantity, and diversity is increasing steadily, which is attracting global attention for the development of new recombinant protein production technologies and the engineering of conventional established expression systems based on bacteria or mammalian cell cultures. Since the advancements of plant genetic engineering in the 1980s, plants have been used for the production of economically valuable, biologically active non-native proteins or biopharmaceuticals, the concept termed as plant molecular farming (PMF). PMF is considered as a cost-effective technology that has grown and advanced tremendously over the past two decades. The development and improvement of the transient expression system has significantly reduced the protein production timeline and greatly improved the protein yield in plants. The major factors that drive the plant-based platform towards potential competitors for the conventional expression system are cost-effectiveness, scalability, flexibility, versatility, and robustness of the system. Many biopharmaceuticals including recombinant vaccine antigens, monoclonal antibodies, and other commercially viable proteins are produced in plants, some of which are in the pre-clinical and clinical pipeline. In this review, we consider the importance of a plant- based production system for recombinant protein production, and its potential to produce biopharmaceuticals is discussed.

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