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Yoko Shoji,
R Mark Jones,
Vadim Mett,
Jessica A Chichester,
Konstantin Musiychuk,
Xiangjie Sun,
Terrence M Tumpey,
Brian J Green,
Moneim Shamloul,
Joey Norikane, Hong Bi,
Caitlin E Hartman,
Cory Bottone,
Michelle Stewart,
Stephen J Streatfield,
Vidadi Yusibov
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ABSTRACT: The increased worldwide awareness of seasonal and pandemic influenza, including pandemic H1N1 virus, has stimulated interest in the development of economic platforms for rapid, large-scale production of safe and effective subunit vaccines. In recent years, plants have demonstrated their utility as such a platform and have been used to produce vaccine antigens against various infectious diseases. Previously, we have produced in our transient plant expression system a recombinant monomeric hemagglutinin (HA) protein (HAC1) derived from A/California/04/09 (H1N1) strain of influenza virus and demonstrated its immunogenicity and safety in animal models and human volunteers. In the current study, to mimic the authentic HA structure presented on the virus surface and to improve stability and immunogenicity of the HA antigen, we generated trimeric HA by introducing a trimerization motif from a heterologous protein into the HA sequence. Here, we describe the engineering, production in Nicotiana benthamiana plants, and characterization of the highly purified recombinant trimeric HA protein (tHA-BC) from A/California/04/09 (H1N1) strain of influenza virus. The results demonstrate the induction of serum hemagglutination inhibition antibodies by tHA-BC and its protective efficacy in mice against a lethal viral challenge, at levels comparable with the licensed egg-based H1N1 influenza vaccine. In addition, the immunogenic and protective doses of tHA-BC were much lower compared to monomeric HAC1. Further investigation into the optimum vaccine dose and/or regimen as well as the stability of trimerized HA is necessary to determine whether trimeric HA is a more potent vaccine antigen than monomeric HA.
Human vaccines & immunotherapeutics. 01/2013; 9(3).
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Yoko Shoji,
Christine E Farrance,
James Bautista, Hong Bi,
Konstantin Musiychuk,
April Horsey,
Heewoo Park,
Jennifer Jaje,
Brian J Green,
Moneim Shamloul,
Satish Sharma,
Jessica A Chichester,
Vadim Mett,
Vidadi Yusibov
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ABSTRACT: Influenza virus is a globally important respiratory pathogen that causes a high degree of annual morbidity and mortality. Significant antigenic drift results in emergence of new, potentially pandemic, virus variants. The best prophylactic option for controlling emerging virus strains is to manufacture and administer pandemic vaccines in sufficient quantities and to do so in a timely manner without impacting the regular seasonal influenza vaccine capacity. Current, egg-based, influenza vaccine production is well established and provides an effective product, but has limited capacity and speed.
To satisfy the additional global demand for emerging influenza vaccines, high-performance cost-effective technologies need to be developed. Plants have a potential as an economic and efficient large-scale production platform for vaccine antigens.
In this study, a plant virus-based transient expression system was used to produce hemagglutinin (HA) proteins from the three vaccine strains used during the 2008-2009 influenza season, A/Brisbane/59/07 (H1N1), A/Brisbane/10/07 (H3N2), and B/Florida/4/06, as well as from the recently emerged novel H1N1 influenza A virus, A/California/04/09.
The recombinant plant-based HA proteins were engineered and produced in Nicotiana benthamiana plants within 2 months of obtaining the genetic sequences specific to each virus strain. These antigens expressed at the rate of 400-1300 mg/kg of fresh leaf tissue, with >70% solubility. Immunization of mice with these HA antigens induced serum anti-HA IgG and hemagglutination inhibition antibody responses at the levels considered protective against these virus infections.
These results demonstrate the feasibility of our transient plant expression system for the rapid production of influenza vaccine antigens.
Influenza and Other Respiratory Viruses 10/2011; 6(3):204-10. · 4.16 Impact Factor
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Yoko Shoji,
Jessica A Chichester,
Mark Jones,
Slobodanka D Manceva,
Emily Damon,
Vadim Mett,
Konstantin Musiychuk, Hong Bi,
Christine Farrance,
Moneim Shamloul,
Natasha Kushnir,
Satish Sharma,
Vidadi Yusibov
[show abstract]
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ABSTRACT: In 2009, a novel H1N1 swine influenza virus was isolated from infected humans in Mexico and the United States, and rapidly spread around the world. Another virus, a highly pathogenic avian influenza virus of the H5N1 subtype, identified by the World Health Organization as a potential pandemic threat in 1997, continues to be a significant risk. While vaccination is the preferred strategy for the prevention and control of influenza infections, the traditional egg-based approach to producing influenza vaccines does not provide sufficient capacity and adequate speed to satisfy global needs to combat newly emerging strains, seasonal or potentially pandemic. Significant efforts are underway to develop and implement new cell substrates with improved efficiency for influenza vaccine development and manufacturing. In recent years, plants have been used to produce recombinant proteins including subunit vaccines and antibodies. The main advantages of using plant systems for the production of vaccine antigens against influenza are their independence from pathogenic viruses, and cost and time efficiency. Here, we describe the large-scale production of recombinant hemagglutinin proteins from A/California/04/09 (H1N1) and A/Indonesia/05/05 (H5N1) strains of influenza virus in Nicotiana benthamiana plants, and their immunogenicity (serum hemagglutination inhibition and virus neutralizing antibodies), and safety in animal models. These results support the testing of these candidate vaccines in human volunteers and also the utility of our plant expression system for large-scale recombinant influenza vaccine production.
Human vaccines 01/2011; 7 Suppl:41-50. · 3.58 Impact Factor
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Vadim Mett,
Jessica A Chichester,
Michelle L Stewart,
Konstantin Musiychuk, Hong Bi,
Carolyn J Reifsnyder,
Anna K Hull,
Mark T Albrecht,
Stanley Goldman,
Les W J Baillie,
Vidadi Yusibov
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ABSTRACT: The health and economic burden of infectious diseases in general and bioterrorism in particular necessitate the development of medical countermeasures. One proven approach to reduce the disease burden and spread of pathogen is treatment with monoclonal antibodies (mAb). mAbs can prevent or reduce severity of the disease by variety of mechanisms, including neutralizing pathogen growth, limiting its spread from infected to adjacent cells, or by inhibiting biological activity of toxins, such as anthrax lethal toxin. Here, we report the production of glycosylated (pp-mAb (PA) ) and non-glycosylated (pp-mAb (PANG) ) versions of a plant-derived mAb directed against protective antigen (PA) of Bacillus anthracis in Nicotiana benthamiana plants using agroinfiltration. Both forms of the antibody were able to neutralize anthrax lethal toxin activity in vitro and protect mice against an intraperitoneal challenge with spores of B. anthracis Sterne strain. A single 180 µg intraperitoneal dose of pp-mAb (PA) or pp-mAb (PANG) provided 90% and 100% survival, respectively. When tested in non-human primates, pp-mAb (PANG) was demonstrated to be superior to pp-mAb (PA) in that it had a significantly longer terminal half-life and conferred 100% protection against a lethal dose of aerosolized anthrax spore challenge after a single 5 mg/kg intravenous dose compared to a 40% survival rate conferred by pp-mAb (PA) . This study demonstrates the potential of a plant-produced non-glycosylated antibody as a useful tool for the treatment of inhalation anthrax.
Human vaccines 01/2011; 7 Suppl:183-90. · 3.58 Impact Factor
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Yoko Shoji,
Christine E Farrance, Hong Bi,
Moneim Shamloul,
Brian Green,
Slobodanka Manceva,
Amy Rhee,
Natalia Ugulava,
Gourgopal Roy,
Konstantin Musiychuk,
Jessica A Chichester,
Vadim Mett,
Vidadi Yusibov
[show abstract]
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ABSTRACT: Highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype have been identified as a potential pandemic threat by the World Health Organization (WHO). Since 1997, these viruses have been spreading from Asia to Europe and Africa with increasing genetic and antigenic diversities. Vaccination is the preferred strategy for the prevention and control of influenza infections and the availability of a system for the rapid engineering and production of vaccines is required in the event of an influenza pandemic. In this study, we engineered and produced recombinant hemagglutinin (HA) from A/Bar-headed Goose/Qinghai/1A/05 (clade 2.2) and A/Anhui/1/2005 (clade 2.3) in Nicotiana benthamiana plants. Immunization of mice with these plant-derived HA antigens elicited serum hemagglutination inhibition (HI) and virus neutralization (VN) antibodies. These results suggest the utility of our plant-expression system for recombinant influenza vaccine production.
Vaccine 03/2009; 27(25-26):3467-70. · 3.77 Impact Factor
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Yoko Shoji, Hong Bi,
Konstantin Musiychuk,
Amy Rhee,
April Horsey,
Gourgopal Roy,
Brian Green,
Moneim Shamloul,
Christine E Farrance,
Barbara Taggart,
Nutan Mytle,
Natalia Ugulava,
Shailaja Rabindran,
Vadim Mett,
Jessica A Chichester,
Vidadi Yusibov
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ABSTRACT: The global spread of highly pathogenic avian influenza virus (H5N1 subtype) has promoted efforts to develop human vaccines against potential pandemic outbreaks. However, current platforms for influenza vaccine production are cumbersome, limited in scalability and often require the handling of live infectious virus. We describe the production of hemagglutinin from the A/Indonesia/05/05 strain of H5N1 influenza virus by transient expression in plants, and demonstrate the immunogenicity and protective efficacy of the vaccine candidate in animal models. Immunization of mice and ferrets with plant-derived hemagglutinin elicited serum hemagglutinin-inhibiting antibodies and protected the ferrets against challenge infection with a homologous virus. This demonstrates that plant-derived H5 HA is immunogenic in mice and ferrets, and can induce protective immunity against infection with highly pathogenic avian influenza virus. Plants could therefore be suitable as a platform for the rapid, large-scale production of influenza vaccines in the face of a pandemic.
Vaccine 01/2009; 27(7):1087-92. · 3.77 Impact Factor
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Yoko Shoji,
Jessica A Chichester, Hong Bi,
Konstantin Musiychuk,
Patricia de la Rosa,
Lauren Goldschmidt,
April Horsey,
Natalia Ugulava,
Gene A Palmer,
Vadim Mett,
Vidadi Yusibov
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ABSTRACT: Influenza is a globally important respiratory pathogen that causes a high degree of morbidity and mortality annually. Although current vaccines are effective against virus infection, new strategies need to be developed to satisfy the global demand for an influenza vaccine. To address this point, we have engineered and produced the full-length hemagglutinin (HA) protein from the A/Wyoming/03/03 (H3N2) strain of influenza in plants. The antigenicity of this plant-produced HA was confirmed by ELISA and single-radial immunodiffusion (SRID) assays. Immunization of mice with plant-produced HA resulted in HA-specific humoral (IgG1, IgG2a and IgG2b) and cellular (IFNgamma and IL-5) immune responses. In addition, significant serum hemagglutination inhibition (HI) and virus neutralizing (VN) antibody titers were obtained with an antigen dose as low as 5mug. These results demonstrate that plant-produced HA protein is antigenic and can induce immune responses in mice that correlate with protection.
Vaccine 07/2008; 26(23):2930-4. · 3.77 Impact Factor
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Vadim Mett,
Konstantin Musiychuk, Hong Bi,
Christine E Farrance,
April Horsey,
Natalia Ugulava,
Yoko Shoji,
Patricia de la Rosa,
Gene A Palmer,
Shailaja Rabindran,
Stephen J Streatfield,
Alison Boyers,
Michael Russell,
Alex Mann,
Robert Lambkin,
John S Oxford,
Geoffrey C Schild,
Vidadi Yusibov
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ABSTRACT: Influenza A viruses are of major concern for public health, causing worldwide epidemics associated with high morbidity and mortality. Vaccines are critical for protection against influenza, but given the recent emergence of new strains with pandemic potential, and some limitations of the current production systems, there is a need for new approaches for vaccine development.
To demonstrate the immunogenicity and protective efficacy of plant-produced influenza antigens. Method We engineered, using influenza A/Wyoming/3/03 (H3N2) as a model virus, the stem and globular domains of hemagglutinin (HA) produced in plants as fusions to a carrier protein and used purified antigens with and without adjuvant for ferret immunization.
These plant-produced antigens were highly immunogenic and conferred complete protection against infection in the ferret challenge model. The addition of plant-produced neuraminidase was shown to enhance the immune response in ferrets.
Plants can be used as a production vehicle for vaccine development against influenza. Domains of HA can generate protective immune responses in ferrets.
Influenza and Other Respiratory Viruses 02/2008; 2(1):33-40. · 4.16 Impact Factor
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Konstantin Musiychuk,
Natalie Stephenson, Hong Bi,
Christine E Farrance,
Goran Orozovic,
Maria Brodelius,
Peter Brodelius,
April Horsey,
Natalia Ugulava,
Abdel-Moneim Shamloul,
Vadim Mett,
Shailaja Rabindran,
Stephen J Streatfield,
Vidadi Yusibov
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ABSTRACT: Historically, most vaccines have been based on killed or live-attenuated infectious agents. Although very successful at immunizing populations against disease, both approaches raise safety concerns and often have limited production capacity. This has resulted in increased emphasis on the development of subunit vaccines. Several recombinant systems have been considered for subunit vaccine manufacture, including plants, which offer advantages both in cost and in scale of production. We have developed a plant expression system utilizing a 'launch vector', which combines the advantageous features of standard agrobacterial binary plasmids and plant viral vectors, to achieve high-level target antigen expression in plants. As an additional feature, to aid in target expression, stability and purification, we have engineered a thermostable carrier molecule to which antigens are fused. We have applied this launch vector/carrier system to engineer and express target antigens from various pathogens, including, influenza A/Vietnam/04 (H5N1) virus.
Influenza and Other Respiratory Viruses 02/2007; 1(1):19-25. · 4.16 Impact Factor
