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Stephen J Streatfield

Publications and source records attributed to Stephen J Streatfield.

15 recordsLinked to original sources
PubMed2007

Approaches to achieve high-level heterologous protein production in plants.

Plants offer an alternative to microbial fermentation and animal cell cultures for the production of recombinant proteins. For protein pharmaceuticals, plant systems are inherently safer than native and even recombinant animal sources. In addition, post-translational modifications, such as glycosylation, which cannot be achieved with bacterial fermentation, can be accomplished using plants. The main advantage foreseen for plant systems is reduced production costs. Plants should have a particular advantage for proteins produced in bulk, such as industrial enzymes, for which product pricing is low. In addition, edible plant tissues are well suited to the expression of vaccine antigens and pharmaceuticals for oral delivery. Three approaches have been followed to express recombinant proteins in plants: expression from the plant nuclear genome; expression from the plastid genome; and expression from plant tissues carrying recombinant plant viral sequences. The most important factor in moving plant-produced heterologous proteins from developmental research to commercial products is to ensure competitive production costs, and the best way to achieve this is to boost expression. Thus, considerable research effort has been made to increase the amount of recombinant protein produced in plants. This research includes molecular technologies to increase replication, to boost transcription, to direct transcription in tissues suited for protein accumulation, to stabilize transcripts, to optimize translation, to target proteins to subcellular locations optimal for their accumulation, and to engineer proteins to stabilize them. Other methods include plant breeding to increase transgene copy number and to utilize germplasm suited to protein accumulation. Large-scale commercialization of plant-produced recombinant proteins will require a combination of these technologies.

Plant Physiological Phenomena↗
PubMed2006

Engineered chloroplasts as vaccine factories to combat bioterrorism.

Bacillus anthracis is ideal for making biological weapons, but the licensed anthrax vaccine is unsuitable for widespread public administration. Recombinant subunit-vaccine candidates offer potential alternatives, and plant-based production systems facilitate the inexpensive bulking of target antigens. A recent report demonstrates expression of anthrax protective antigen in tobacco chloroplasts--this material is immunogenic and protective when injected into mice. Provided an economic purification scheme can be developed, this technology holds promise for an improved vaccine.

Anthrax↗
PubMed2006

Mucosal immunization using recombinant plant-based oral vaccines.

The induction of mucosal immunity is very important in conferring protection against pathogens that typically invade via mucosal surfaces. Delivery of a vaccine to a mucosal surface optimizes the induction of mucosal immunity. The apparent linked nature of the mucosal immune system allows delivery to any mucosal surface to potentially induce immunity at others. Oral administration is a very straightforward and inexpensive approach to deliver a vaccine to the mucosal lining of the gut. However, vaccines administered by this route are subject to proteolysis in the gastrointestinal tract. Thus, dose levels for protein subunit vaccines are likely to be very high and the antigen may need to be protected from proteolysis for oral delivery to be efficacious. Expression of candidate vaccine antigens in edible recombinant plant material offers an inexpensive means to deliver large doses of vaccines in encapsulated forms. Certain plant tissues can also stably store antigens for extensive periods of time at ambient temperatures, obviating the need for a cold-chain during vaccine storage and distribution, and so further limiting costs. Antigens can be expressed from transgenes stably incorporated into a host plant's nuclear or plastid genome, or from engineered plant viruses infected into plant tissues. Molecular approaches can serve to boost expression levels and target the expressed protein for appropriate post-translational modification. There is a wide range of options for processing plant tissues to allow for oral delivery of a palatable product. Alternatively, the expressed antigen can be enriched or purified prior to formulation in a tablet or capsule for oral delivery. Fusions to carrier molecules can stabilize the expressed antigen, aid in antigen enrichment or purification strategies, and facilitate delivery to effector sites in the gastrointestinal tract. Many antigens have been expressed in plants. In a few cases, vaccine candidates have entered into early phase clinical trials, and in the case of farmed animal vaccines into relevant animal trials.

Administration, Oral↗
PubMed2005

Oral hepatitis B vaccine candidates produced and delivered in plant material.

Hepatitis B is a major global health problem; approximately two billion people are infected with the virus worldwide, despite the fact that safe and efficacious vaccines have been developed and used for nearly 20 years. Prohibitive costs for vaccine purchase and administration restrict uptake in many developing nations. Agencies such as the Global Alliance for Vaccination and Immunization are helping to make current vaccines more available, but reduced costs would greatly aid this effort. Oral delivery is an option to reduce the expense of administering hepatitis B vaccines. It may also improve compliance, and orally delivered vaccines may be more efficacious among poor responders to current vaccines. However, to induce protective efficacy, oral administration may require encapsulation of antigen and delivery of large doses. Plant-based expression systems offer an oral delivery alternative with low production costs, and they also encapsulate the antigen. Some plant-based systems also stabilize antigen and therefore reduce storage and distribution costs. The hepatitis B major surface antigen has been expressed in several plant systems. A variety of regulatory sequences and subcellular targets have been used to achieve expression suitable for early stage clinical trials. However, further increase in expression will be necessary for practical and efficacious products. Appropriate processing can yield palatable products with uniform antigen concentration. The antigen expressed in plant systems shows extensive disulphide cross-linking and oligomerization and forms virus-like particles. Oral delivery of the antigen in plant material can induce a serum antibody response, prime the immune system for a subsequent injection of antigen and give a boosted response to a prior injection. Small scale clinical trials in which the antigen has been delivered orally in edible plant material indicate safety and immunogenicity.

Animals↗
PubMed2005

Expression of the sweet protein brazzein in maize for production of a new commercial sweetener.

The availability of foods low in sugar content yet high in flavour is critically important to millions of individuals conscious of carbohydrate intake for diabetic or dietetic purposes. Brazzein is a sweet protein occurring naturally in a tropical plant that is impractical to produce economically on a large scale, thus limiting its availability for food products. We report here the use of a maize expression system for the production of this naturally sweet protein. High expression of brazzein was obtained, with accumulation of up to 4% total soluble protein in maize seed. Purified corn brazzein possessed a sweetness intensity of up to 1200 times that of sucrose on a per weight basis. In addition, application tests demonstrated that brazzein-containing maize germ flour could be used directly in food applications, providing product sweetness. These results demonstrate that high-intensity sweet protein engineered into food products can give sweetener attributes useful in the food industry.

Journal Article↗
PubMed2005

Delivery of plant-derived vaccines.

Many protein subunit vaccine candidates have been expressed in transgenic plants, and in a few cases the recombinant material has entered early phase clinical or target animal trials. The expressed protein can be purified prior to formulation for any preferred delivery approach. However, there are major cost advantages associated with avoiding protein purification and pursuing the oral delivery of a processed plant product containing the recombinant protein. Grains and dry products that are processed from fresh plant tissues can stably store expressed proteins for extended periods of time at room temperature, making refridgeration unnecessary during storage and distribution. Encapsulation of recombinant proteins in plant tissues guards against their rapid degradation in the gut, therefore facilitating the uptake and induction of appropriate immune responses. Early trial data with plant-based vaccine candidates has shown promising safety and efficacy.

Administration, Oral↗
PubMed2005

Regulatory issues for plant-made pharmaceuticals and vaccines.

Recombinant plant systems potentially offer economic alternatives to produce large amounts of pharmaceutical proteins, including those used in subunit vaccines. Plant systems also provide a convenient oral delivery option, overcoming the cost and inconvenience of purification and injections. The production of pharmaceutical proteins in transgenic plants is tightly regulated, with the US Department of Agriculture focusing on containment of recombinant material and the US Food and Drug Administration focusing on the production system as it relates to manufacture of the drug or vaccine. Current regulations for the production of plant-made pharmaceuticals are to prevent recombinant proteins from entering the food chain or from persisting in the environment, and to guard against recombinant nucleic acid sequences entering genomes of food or feed crops, or wild species. Several alternative plant production systems are being developed. Each system has its strengths and weaknesses with regard to the economics of production, options for alternative routes of administration, authenticity of products and ease with which the production system can be contained. Risk assessments can be used as a means to quantify risks of inadvertent human or environmental exposure to plant-made pharmaceuticals. Several technologies are being tested that reduce the probability of plant-made pharmaceuticals, or genes encoding them, escaping production sites.

Animals↗
PubMed2004

A corn-based delivery system for animal vaccines: an oral transmissible gastroenteritis virus vaccine boosts lactogenic immunity in swine.

Recombinant plant expression systems offer a means to produce large quantities of selected antigens for subunit vaccines. Cereals are particularly well-suited expression vehicles since the expressed proteins can be stored at relatively high concentrations for extended periods of time without degradation and dry seed can be formulated into oral vaccines suitable for commercial applications. A subunit vaccine candidate directed against porcine transmissible gastroenteritis virus and expressed in corn seed has been developed for oral delivery to swine. Here, we show that this vaccine, when administered to previously sensitized gilts, can boost neutralizing antibody levels in the animals' serum, colostrum and milk. Thus, this vaccine candidate is effective at boosting lactogenic immunity and is appropriate to pursue through large-scale field trials preceding commercialization.

Administration, Oral↗
PubMed2004

Analysis of the maize polyubiquitin-1 promoter heat shock elements and generation of promoter variants with modified expression characteristics.

The maize polyubiquitin-1 (Ubi-1) promoter is one of a few select promoters used to express foreign genes in monocots, such that recombinant proteins can be produced at commercially viable levels. Modifying the activity, specificity and responsiveness of such promoters provides a means to achieve desired levels and patterns of expression of genes encoding target products. Ubi-1 is constitutively expressed but is further induced by heat shock. The promoter contains two overlapping sequences with similarity to defined heat shock elements and we show that these sequences are also present upstream of the Ubi-1 homologue isolated from teosinte. Both the maize and teosinte promoters can mediate a heat shock response in transgenic maize. We have dissected the overlapping maize Ubi-1 promoter heat shock elements and demonstrate that the 3' element is required to mediate a heat shock response. The Ubi-1 promoter is particularly active in tissues consisting of rapidly dividing cells, and within the seed it is strongly biased towards driving expression in the embryo. However, replacement of the heat shock elements with a trimer of a basic domain/leucine zipper factor binding site of a pea lectin promoter shifts the balance in seed expression towards the endosperm. The Ubi-1 variants described here differ in their overall activity in the seed, but they all show potential for driving high levels of heterologous gene expression in maize.

Base Sequence↗
PubMed2003

Corn as a production system for human and animal vaccines.

The synthesis of selected antigens in plants and their oral delivery has great potential for reducing the costs of vaccine production and administration. The application of this technology requires antigen concentrations in final plant material to be uniform to ensure consistent dosing. In addition, antigen levels should be such as to allow the volume of each dose, containing a set amount of antigen, to be practical for oral delivery. Here, we demonstrate that the Lt-B protein of enterotoxigenic E. coli is evenly distributed in defatted corn germ prepared from transgenic grain. Furthermore, the choice of sub-cellular location for Lt-B affects accumulation of the protein in excess of four orders of magnitude.

Bacterial Toxins↗
PubMed2003

Plant-based vaccines.

Plant systems are reviewed with regard to their ability to express and produce subunit vaccines. Examples of different types of expression systems producing a variety of vaccine candidates are illustrated. Many of these subunit vaccines have been purified and shown to elicit an immune response when injected into animal models. This review also includes vaccines that have been administered orally in a non-purified form as a food or feed product. Cases are highlighted which demonstrate that orally delivered plant-based vaccines can elicit immune responses and in some case studies, confer protection. Examples are used to illustrate some of the inherent advantages of a plant-based system, such as cost, ease of scale-up and convenience of delivery. Also, some of the key steps are identified that will be necessary to bring these new vaccines to the market.

Administration, Oral↗
PubMed2003

Advantageous features of plant-based systems for the development of HIV vaccines.

Plants have recently become an attractive option for the production of recombinant proteins. Plant-based systems can be used to produce many classes of foreign proteins including candidate vaccine antigens. The selected antigen can be purified from plant material prior to delivery by the preferred route, or alternatively delivered orally in edible plant material that has been processed to give a homogeneous and stable product. Several plant species have been used to express a wide range of vaccine candidates with tobacco, potato and corn being particularly favored. Corn seed is especially well suited to various food processing technologies that generate dry homogeneous material suitable for extended storage and refrigeration-free transport and distribution. Many antigens have been expressed in corn and assessed for efficacy in trials with generally positive results. Candidate HIV vaccines are particularly good targets for plant-based oral delivery since there is a great need for an easily distributed affordable vaccine that could be administered without injection and induce strong mucosal immune responses. As a first step in evaluating plant expression technology with a relevant antigen that might easily be tested in an animal system, we expressed the SIV major surface glycoprotein gp130 (analogous to HIV gp120) in corn seed. Expression levels were achieved that are compatible with conducting oral delivery trials in animals.

AIDS Vaccines↗
PubMed2003

Plant production systems for vaccines.

Plants offer an attractive alternative for the production and delivery of subunit vaccines. Various antigens have been expressed at sufficiently high levels in plants to render vaccine development practical. An increasing body of evidence demonstrates that these plant-produced antigens can induce immunogenic responses and confer protection when delivered orally. Plant-based vaccines are relatively inexpensive to produce and production can be rapidly scaled up. There is also the potential for oral delivery of these vaccines, which can dramatically reduce distribution and delivery costs. Here we describe the technology to develop plant-based vaccines, review their advantages and discuss potential roadblocks and concerns over their commercialization. We also speculate on likely future developments with these vaccines and on their potential impact in the realms of human and animal health.

Administration, Oral↗
PubMed2002

Delivery of subunit vaccines in maize seed.

The use of recombinant gene technologies by the vaccine industry has revolutionized the way antigens are generated, and has provided safer, more effective means of protecting animals and humans against bacterial and viral pathogens. Viral and bacterial antigens for recombinant subunit vaccines have been produced in a variety of organisms. Transgenic plants are now recognized as legitimate sources for these proteins, especially in the developing area of oral vaccines, because antigens have been shown to be correctly processed in plants into forms that elicit immune responses when fed to animals or humans. Antigens expressed in maize (Zea mays) are particularly attractive since they can be deposited in the natural storage vessel, the corn seed, and can be conveniently delivered to any organism that consumes grain. We have previously demonstrated high level expression of the B-subunit of Escherichia coli heat-labile enterotoxin and the spike protein of swine transmissible gastroenteritis in corn, and have demonstrated that these antigens delivered in the seed elicit protective immune responses. Here we provide additional data to support the potency, efficacy, and stability of recombinant subunit vaccines delivered in maize seed.

Administration, Oral↗