Expression of H5 hemagglutinin vaccine antigen in common duckweed (Lemna minor) protects against H5N1 high pathogenicity avian influenza virus challenge in immunized chickens

Bertran, Kateri; Thomas, Colleen J.; Guo, Xuan; Bublot, Michel; Pritchard, Nikki; Regan, Jeffrey T.; Cox, Kevin M.; Gasdaska, John R.; Dickey, Lynn F.; Kapczynski, Darrell R.; Swayne, David E.

doi:10.1016/j.vaccine.2015.05.076

Cited by 38 publications

(16 citation statements)

References 46 publications

(58 reference statements)

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“…Efficient nuclear transformation protocols for two species of duckweed, L. gibba and L. minor, were established in 2001 [121], and >20 recombinant proteins were produced with expression levels up to 25% of TSP [32]. Products include industrial enzymes, e.g., E1 endoglucanase [122], and many therapeutic proteins, e.g., mAb [123], plasminogen [124], interferon α2 [125], vaccine antigen avian influenza including H5N1 hemagglutinin [126,127], and M2e peptide [117].…”

Section: Duckweedmentioning

confidence: 99%

Platforms for Plant-Based Protein Production

Xu¹,

Towler²,

Weathers³

2016

Bioprocessing of Plant in Vitro Systems

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Plant molecular farming depends on a diversity of plant systems for production of useful recombinant proteins. These proteins include protein biopolymers, industrial proteins and enzymes, and therapeutic proteins. Plant production systems include microalgae, cells, hairy roots, moss, and whole plants with both stable and transient expression. Production processes involve a narrowing diversity of bioreactors for cell, hairy root, microalgae, and moss cultivation. For whole plants, both field and automated greenhouse cultivation methods are used with

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Section: Duckweedmentioning

confidence: 99%

Platforms for Plant-Based Protein Production

Xu¹,

Towler²,

Weathers³

2016

Bioprocessing of Plant in Vitro Systems

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“…The main targets for subunit vaccine development have been its principal surface antigen hemagglutinin (HA), which plays a major role in functional activity of the virus and is also a main target for neutralizing antibodies of the host, and the M2e viral peptide, the extracellular domain of the viral protein M2, which is highly conserved between various influenza strains and therefore a promising candidate for a universal influenza vaccine. H5N1 HA has been expressed in N. benthamiana in full-length (Kalthoff et al, 2010) and in the form of VLPs (Landry et al, 2010), in Lotus corniculatus (Guo et al, 2012) and most recently, duckweed (Bertran et al, 2015). In addition, recombinant HA from A/Bar-headed Goose/Qinghai/1A/05 (clade 2.2) and A/Anhui/1/2005 (clade 2.3) have been expressed in N. benthamiana (Shoji et al, 2009).…”

Section: Avian Influenza Virus H5n1mentioning

confidence: 99%

“…Recently, synthetic HA gene from H5N1 virus A/chicken/Indonesia/7/2003 (Indo/03) was expressed in duckweed. Its efficacy was first tested in birds immunized with 0.2 µg or 2.3 µg HA and challenged with 10 6 mean chicken embryo infectious doses (EID50) of homologous virus strain (Bertran et al, 2015) and in birds immunized with 0.9 µg or 2.2 µg HA challenged with 10 6 EID50 of heterologous H5N1 virus strains A/chicken/Vietnam/NCVD-421/2010 (VN/10) or A/chicken/West Java/PWT-WIJ/2006 (PWT/06) (Bertran et al, 2015). Almost all birds immunized with 0.2 or 2.3 µg of HA elicited anti-Indo/03 antibodies and were protected against homologous virus challenge, 100 % of birds immunized with either dosage of HA showed protection against VN/10 challenge while birds challenged with PWT/06 showed 50 % mortality when immunized with 0.9 µg HA and 30 % mortality when immunized with 2.2 µg HA.…”

Section: Avian Influenza Virus H5n1mentioning

confidence: 99%

Production of vaccines for treatment of infectious diseases by transgenic plants

Ledl

Luthar

2016

AAS

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Since the first pathogen antigen was expressed in transgenic plants with the aim of producing edible vaccine in early 1990s, transgenic plants have become a well-established expression system for production of alternative vaccines against various human and animal infectious diseases. The main focus of plant expression systems in the last five years has been on improving expression of well-studied antigens such as porcine reproductive and respiratory syndrome (PRRSV), bovine viral diarrhea disease virus (BVDV), footh and mouth disease virus (FMDV), hepatitis B surface antigen (HBsAg), rabies G protein, rotavirus, Newcastle disease virus (NDV), Norwalk virus capsid protein (NVCP), avian influenza virus H5N1, Escherichia coli heat-labile enterotoxin subunit B (LT-B), cholera toxin B (CT-B), human immunodeficiency virus (HIV), artherosclerosis, ebola and anthrax. Significant increases in expression have been obtained using improved expression vectors, different plant species and transformation methods.Key words: transgenic plants; vaccine production; recombinant proteins; antibodies; infectious diseases; risk assessment IZVLEČEK ZDRAVLJENJE NALEZLJIVIH BOLEZNI S CEPIVI PRIDOBLJENIMI S TRANSGENIMI RASTLINAMIPridobivanje aktivnih sestavin za cepiva s transgenimi rastlinami se je začelo v devetdesetih letih prejšnjega stoletja. Od takrat se transgene rastline uveljavljajo kot alternativni ekspresijski sistem za sintezo aktivnih komponent za cepiva proti številnim humanim in živalskim nalezljivim boleznim. V zadnjih petih letih je bil glavni poudarek razvoja namenjen optimizaciji rastlinskih ekspresijskih sistemov za dobro proučene antigene, kot so virus prašičjega respiratornega in reproduktivnega sindroma, virus goveje virusne diareje, virus parkljevke, površinski antigen hepatitisa B, G protein virusa stekline, rotavirus, virus atipične kokošje kuge, plaščni protein Norwalk virusa, sev ptičje gripe H5N1, temperaturno nestabilna B podenota enterotoksina Escherischia coli, kolera toksin B, HIV, arterioskleroza, ebola in antraks. Uporaba optimiziranih transformacijskih metod in ekspresijskih vektorjev v kombinaciji s primerno izbrano rastlinsko vrsto je v večini naštetih primerov vodila do opaznega izboljšanja sinteze posameznih učinkovin.

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“…Several commercially important proteins have already been successfully expressed in duckweed plants: the endoglucanase E1 from Acidothermus cellulolyticus (Sun et al, 2007 ), and monoclonal antibodies against CD20, CD30, and interferon α2b (Cox et al, 2006 ). Protective antigen of swine epidemic diarrhea (Ko et al, 2011 ), tuberculosis antigens ESAT6 and Ag85B (Peterson et al, 2015 ), and hemagglutinin of avian influenza virus H5N1 (Bertran et al, 2015 ; Huong et al, 2015 ) have also been successfully expressed in these plants.…”

Section: Introductionmentioning

confidence: 99%

Expression and Immunogenicity of M2e Peptide of Avian Influenza Virus H5N1 Fused to Ricin Toxin B Chain Produced in Duckweed Plants

et al. 2018

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The amino acid sequence of the extracellular domain of the virus-encoded M2 matrix protein (peptide M2e) is conserved among all subtypes of influenza A strains, enabling the development of a broad-range vaccine against them. We expressed M2e from avian influenza virus A/chicken/Kurgan/5/2005 (H5N1) in nuclear-transformed duckweed plants for further development of an avian influenza vaccine. The 30-amino acid N-terminal fragment of M2, including M2e (denoted M130), was selected for expression. The M2e DNA sequence fused in-frame to the 3′ end of ricin toxin B chain (RTB) was cloned under control of the CaMV 35S promoter into pBI121. The resulting plasmid was used for duckweed transformation, and 23 independent transgenic duckweed lines were obtained. Asialofetuin-binding ELISA of protein samples from the transgenic plants using polyclonal anti-RTB antibodies confirmed the expression of the RTB–M130 fusion protein in 20 lines. Quantitative ELISA of crude protein extracts from these lines showed RTB–M130 accumulation ranging from 0.25–2.5 μg/g fresh weight (0.0006–0.01% of total soluble protein). Affinity chromatography with immobilized asialofetuin and western blot analysis of protein samples from the transgenic plants showed expression of fusion protein RTB–M130 in the aggregate form with a molecular mass of about 70 kDa. Mice were immunized orally with a preparation of total soluble protein from transgenic plants, receiving four doses of 7 μg duckweed-derived RTB–M130 each, with no additional adjuvant. Specific IgG against M2e was detected in immunized mice, and the endpoint titer of nti-M2e IgG was 1,024. It was confirmed that oral immunization with RTB-M130 induces production of specific antibodies against peptide M2e, one of the most conserved antigens of the influenza virus. These results may provide further information for the development of a duckweed-based expression system to produce a broad-range edible vaccine against avian influenza.

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Expression of H5 hemagglutinin vaccine antigen in common duckweed (Lemna minor) protects against H5N1 high pathogenicity avian influenza virus challenge in immunized chickens

Cited by 38 publications

References 46 publications

Platforms for Plant-Based Protein Production

Platforms for Plant-Based Protein Production

Production of vaccines for treatment of infectious diseases by transgenic plants

Expression and Immunogenicity of M2e Peptide of Avian Influenza Virus H5N1 Fused to Ricin Toxin B Chain Produced in Duckweed Plants

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