Arabidopsis thaliana plants expressing Rift Valley fever virus antigens: Mice exhibit systemic immune responses as the result of oral administration of the transgenic plants

Kalbina, Irina; Lagerqvist, Nina; Moiane, Belisário; Ahlm, Clas; Andersson, Sören; Strid, Åke; Falk, Kerstin I.

doi:10.1016/j.pep.2016.07.003

Cited by 14 publications

(8 citation statements)

References 38 publications

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“…In the current study, use of 197 g of agroinfiltrated leaves resulted in N-protein yields ranging from 500 to 558 μg/g fresh leaf material. These yields are much higher than those previously obtained in transgenic A. thaliana (3.8 μg/g of fresh leaf weight) [33], and comparable to those achieved for recombinantly-produced N-protein in E. coli of 0.34–0.71 mg/mL [10, 16]. Moreover, sufficient yields of soluble plant-produced N-protein were easily purified for testing.…”

Section: Discussionsupporting

confidence: 52%

“…In addition, a plant production system is free of contaminating animal-derived agents, which is often a major concern; it is safe to handle during production, more cost effective than other expression platforms and allows for rapid, scalable production of recombinant proteins ([28–32], Merlin, 2014 #225). RVFV N-protein has recently been expressed in transgenic Arabidopsis thaliana [33]: however, this was intended for use as a vaccine candidate and yields were relatively low, ranging from levels of 3.3 μg/g in root tissue to 3.8 μg/g fresh weight in leaves.…”

Section: Introductionmentioning

confidence: 99%

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Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen

et al. 2018

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BackgroundRift Valley fever virus (RVFV), the causative agent of Rift Valley fever, is an enveloped single-stranded negative-sense RNA virus in the genus Phlebovirus, family Bunyaviridae. The virus is spread by infected mosquitoes and affects ruminants and humans, causing abortion storms in pregnant ruminants, high neonatal mortality in animals, and morbidity and occasional fatalities in humans. The disease is endemic in parts of Africa and the Arabian Peninsula, but is described as emerging due to the wide range of mosquitoes that could spread the disease into non-endemic regions.There are different tests for determining whether animals are infected with or have been exposed to RVFV. The most common serological test is antibody ELISA, which detects host immunoglobulins M or G produced specifically in response to infection with RVFV. The presence of antibodies to RVFV nucleocapsid protein (N-protein) is among the best indicators of RVFV exposure in animals. This work describes an investigation of the feasibility of producing a recombinant N-protein in Nicotiana benthamiana and using it in an ELISA.ResultsThe human-codon optimised RVFV N-protein was successfully expressed in N. benthamiana via Agrobacterium-mediated infiltration of leaves. The recombinant protein was detected as monomers and dimers with maximum protein yields calculated to be 500–558 mg/kg of fresh plant leaves. The identity of the protein was confirmed by liquid chromatography-mass spectrometry (LC-MS) resulting in 87.35% coverage, with 264 unique peptides. Transmission electron microscopy revealed that the protein forms ring structures of ~ 10 nm in diameter. Preliminary data revealed that the protein could successfully differentiate between sera of RVFV-infected sheep and from sera of those not infected with the virus.ConclusionsTo the best of our knowledge this is the first study demonstrating the successful production of RVFV N-protein as a diagnostic reagent by Agrobacterium-mediated transient heterologous expression in N. benthamiana. Preliminary testing of the antigen showed its ability to distinguish RVFV-positive animal sera from RVFV negative animal sera when used in an enzyme linked immunosorbent assay (ELISA). The cost-effective, scalable and simple production method has great potential for use in developing countries where rapid diagnosis of RVFV is necessary.Electronic supplementary materialThe online version of this article (10.1186/s12896-018-0489-z) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 52%

Section: Introductionmentioning

confidence: 99%

Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen

et al. 2018

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“…To this end, a soluble RVFV Gn glycoprotein has been expressed at low levels in stably transformed Arabidopsis thaliana. Although the recombinant protein could not be detected by Western blotting, mRNA expression was confirmed by RT-PCR and fresh plant material was reported to be orally immunogenic in mice (Kalbina et al, 2016). Our group has expressed and purified a chimeric particle-forming Gn antigen from N. benthamiana plants which contains the transmembrane and cytoplasmic domains of influenza HA (Mbewana, 2017).…”

Section: Other Vector-borne and Zoonotic Virusesmentioning

confidence: 99%

Production of complex viral glycoproteins in plants as vaccine immunogens

Margolin

Chapman

Williamson

et al. 2018

Plant Biotechnology Journal

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SummaryPlant molecular farming offers a cost‐effective and scalable approach to the expression of recombinant proteins which has been proposed as an alternative to conventional production platforms for developing countries. In recent years, numerous proofs of concept have established that plants can produce biologically active recombinant proteins and immunologically relevant vaccine antigens that are comparable to those made in conventional expression systems. Driving many of these advances is the remarkable plasticity of the plant proteome which enables extensive engineering of the host cell, as well as the development of improved expression vectors facilitating higher levels of protein production. To date, the only plant‐derived viral glycoprotein to be tested in humans is the influenza haemagglutinin which expresses at ~50 mg/kg. However, many other viral glycoproteins that have potential as vaccine immunogens only accumulate at low levels in planta. A critical consideration for the production of many of these proteins in heterologous expression systems is the complexity of post‐translational modifications, such as control of folding, glycosylation and disulphide bridging, which is required to reproduce the native glycoprotein structure. In this review, we will address potential shortcomings of plant expression systems and discuss strategies to optimally exploit the technology for the production of immunologically relevant and structurally authentic glycoproteins for use as vaccine immunogens.

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“… 36 A plant-made approach to a RVFV vaccine published recently described the expression in transgenic Arabidopsis thaliana of the N protein and a soluble version of the Gn glycoprotein. 37 Oral dosing of fresh transgenic plant material was immunogenic in mice, and elicited systemic antigen-specific IgG: this augurs well for similar experiments in larger animals. A spur to the deployment of human vaccines for RVFV could be the recent finding that the virus may be involved in human miscarriages in South Sudan.…”

Section: Introductionmentioning

confidence: 72%

Plant-made vaccines and reagents for the One Health initiative

Rybicki

2017

Human Vaccines & Immunotherapeutics

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The One Health initiative is increasingly becoming a prominent discussion topic in animal and human health, with its focus on prevention of spread of zoonotic diseases, both in animals, and from animals to humans. An important part of One Health is that diagnostics and vaccines for diseases may be the same thing – and be used for both humans and animals. One potential problem standing in the way of wider adoption of One Health principles, though, is that use of conventional cell fermentation systems for production of the recombinant proteins that could be used as diagnostics or vaccines is often expensive and is not easily scalable. A solution to this may be the use of plants or plant cells as bioreactors: molecular farming, or the production of biologics in plants, is now a well-established science with many proofs of principle and important proofs of efficacy for especially animal vaccines. This review discusses how molecular farming could enable important advances in One Health, using as examples plant-made vacccines, reagents and therapeutics for influenza viruses, ebolaviruses, rabies virus, bunyaviruses and flaviviruses.

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Arabidopsis thaliana plants expressing Rift Valley fever virus antigens: Mice exhibit systemic immune responses as the result of oral administration of the transgenic plants

Cited by 14 publications

References 38 publications

Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen

Expression of Rift Valley fever virus N-protein in Nicotiana benthamiana for use as a diagnostic antigen

Production of complex viral glycoproteins in plants as vaccine immunogens

Plant-made vaccines and reagents for the One Health initiative

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