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EDIBLE VACCINE - A GREAT BOON IN MEDICINAL SCIENCE

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EDIBLE VACCINE - A GREAT BOON IN MEDICINAL SCIENCE

The genetic material from the microbe responsible for producing cholera toxin into a rice plant, whose genome has recently been sequenced. The plants produced the toxin and when the rice grains were fed to mice they provoked immunity from the diarrhea-causing bacterium. "We are considering rice as a new vaccine production and storage system, and natural vaccine delivery vehicle"The vaccine expressed in rice, or rice-based vaccine, will become a new form of vaccine production and delivery to [the] digestive tract for the initiation of antigen-specific mucosal and systemic immune responses." Rice offers several advantages over traditional vaccines: it does not require needles, purification or refrigeration. In fact, the rice proved just as potent after 18 months of storage at room temperature and the vaccine did not dissolve when exposed to stomach acids, the researchers report in Proceedings of the National Academy of Sciences USA. But the immune response itself will require periodic updating. "Oral boosting should be necessary for the induction of antigenspecific immune response," Because rice grains contain varying amounts of the vaccineroughly 30 micrograms per seeda pill of some kind would need to be created to make sure people get the proper dose. The rice plants would also have to be grown under carefully controlled conditions to ensure appropriate vaccine production. "We do not have any plan to deliver the vaccine as a form of steamed rice, "A powder form of rice-based vaccine will be given in a tablet or capsule form." The researchers point out that because rice plants do not scatter their pollen as widely as some of the other crops genetically modified to produce vaccinescorn, wheat, tomatoesand are widely grown (unlike vaccine-producing bananas and potatoes), they pose less risk of contaminating normal crops and have broader utility. Such rice-based vaccines need not stop at cholera,. The same technique could be used to create rice grains bearing protection against the flu, botulism or anthrax, among other diseases. Someday, the adage may be: A bowl of rice a day, keeps the needle away. Edible vaccine using GM maize in hepatitis B:
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Fig.9: Edible vaccine using GM maize Maize plants to produce a protein used to make the hepatitis B virus vaccine. They hope that their findings could eventually lead to the creation of an edible vaccine that could be locally produced and would dispense with the need for expensive vaccination programmes. More than 2 billion people are infected with hepatitis B, and about 350 million of these are at high risk of serious illness and death from liver damage and liver cancer. A vaccine against the disease is already available, but the Egyptian researchers say that edible vaccines produced by GM plants would be cheaper and would not need to be refrigerated. GM maize plants that produce the protein known as HbsAg, which elicits an immune response against the hepatitis B virus and could be used as a vaccine. To increase the amount of the protein produced by the plants. They have not yet tested the effectiveness of the edible vaccine in animals and humans, but expect that tests will start early next year. Reporting the results at an international conference on genetic engineering, effective vaccine against the disease is vital, as many people are excluded from immunisation programmes because of the expense of the vaccines. Edible vaccine for developing countries that would not require refrigerators for storage, or skilled medical personnel and

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needles to deliver the vaccine. Edible vaccine used in HIV infection:
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RASA LSI

Fig.10: Edible vaccine used in HIV infection HIV is mainly transferred through mucosal sites, which strengthen the rationale of using the oral vaccination route. The Arabidopsis thaliana plants are transformed with a gene emanating from HIV using Agrobacterium tumefaciens-mediated gene transfer. These bacteria, Agrobacterium tumefaciens, spontaneously transfer a part of its genome into the plant genome during infection. This ability is used when the plants are transformed. By first inserting a gene for a HIV protein into Agrobacterium, this bacterium in turn will transfer the HIV gene into the plant. A herbicid resistance gene for selection is transferred at the same time. Only plants that are transformed with these genes can grow on the selective media. The genes transferred to the plants are incorporated randomly into the plant nuclear genome. The research group at rebro Life Science Center has successfully transformed Arabidopsis thaliana and has gone on to produce several transformed plant lines, which express the protein corresponding to the inserted gene. We have also shown that the gene is inherited by the offspring. Hopefully, the protein expressed by these plants will elicit the immune system in the mucosal tissues during consumption. We are now carrying out trials with mice eating these transformed plants in order to establish whether they develop any immunological response. POTATO VACCINE COULD FIGHT STOMACH VIRUS:(60)

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Fig.11: Potato vaccine Vaccine Development have successfully tested a potato-based vaccine to combat the Norwalk Virus, which is spread by contaminated food and water. The virus causes severe abdominal pain and diarrhea. Researchers demonstrated that the edible vaccine is safe and stimulates antibodies, or germ-fighting proteins in the volunteers tested. We are excited about this novel approach to developing vaccines because of its potential to protect individuals around the world, especially in regions where injected vaccines are less practical an edible vaccine could be easy to produce, safe, affordable and effective. To be immunized, people would need to eat a raw vegetable that contains the gene for the vaccine protein. It's exciting to think of the future for this type of vaccine delivery system and the varieties of organisms we may one day be able to fight. The Norwalk Virus and closely related members of the same virus family account for more than 90 percent of non-bacterial gastroenteritis (severe abdominal pain and diarrhea) in developing countries, the rapid and severe spread of the Norwalk Virus is a leading cause of infant mortality. A gene that encoded the Norwalk Virus coat into a potato plant's DNA. The potato vaccine was then tested at the Center for Vaccine Development in a double-blind study of 24 adult volunteers. Results show that 19 of the 20 volunteers (95 percent) who were given the potato containing the vaccine developed antibodies that fight the Norwalk Virus. Edible plant vaccines will continue to be tested in volunteers for the next few years and could be available to protect people in 5-10 years. A potato vaccine against E. coli, a common cause of serious diarrhea in children and adults around the world. The study showed that eating the potato was an effective way to develop immunity to the E. coli toxin. Edible vaccine using tobacco in cervical cancer:
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PHARMACEUTICAL PACKAGING FORMULATION AND EVALUATION OF AMOXICILLIN TRIHYDRATE MODIFIED RELEASE DOSAGE FORMS DESIGN AND DEVELOPMENT OF COLON DRUG DELIVERY SYSTEM OF DICLOFENAC SODIUM DEVELOPMENT OF DOMPERIDONE NASAL GEL USING NATURAL MUCOADHESIVE AGENT OBTAINED FROM THE FRUITS OF DELLINIA INDICA. L. A REVIEW: PARENTERAL

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Fig.12: Edible vaccine using tobacco Human Papillomaviruses (HPV) are the causative agents for cervical cancer, being also involved in skin, head and neck tumors., cervical cancer is one of the main causes of cancer-related deaths. However, Italian researchers have now developed an immunologically active, cost-efficient vaccine against HPV. Currently, more than 150 different types of Human Papillomaviruses are known. The most commonly found HPVs in cervical carcinomas are HPV 16 and 18. The cancer causing factors are the virus proteins E6 and E7, which are known as onco-proteins. Onco-proteins represent a promising target for the development of a therapeutic vaccine against HPV-associated tumors. This has been confirmed by an experiment showing that mice immunized with crude plant extracts containing the onco-protein E7 are partially protected against HPV-induced cancers. Some E7-based HPV vaccines are currently being explored and the first promising results have been disclosed, but they still need further improvement to ensure protection. One method to produce high amounts of E7 protein in a relatively short time is by genetically engineering plants or plant viruses. An advantage of the plant-derived systems is that they generally lack human pathogens, oncogenic DNA sequences and endotoxins This minimizes health risks and lowers the production costs. Plants can be genetically modified by either stable or transient transformation. With stable transformation, the introduced DNA integrates in the genome of the plant, whereas with transient transformation, the introduced gene sequences do not integrate in the genome of the plant. Ideal tools for transient expression are viruses, whose genomes contain, as an additionary gene, the sequences of interest. When the plant is infected with the virus, the virus spreads and replicates and the genes introduced in its genome are expressed in high amounts. The fact that viruses are usually not transmitted by pollen which ensures genetic contamination of wild growing plants is prevented - is a further advantage of viral systems. The HPV16 E7 protein in the cytoplasm of tobacco (Nicotiana benthamiana) plants. To do so, they used the potato virus X (PVX). PVX is a safe tool for genetic engineering, because it does not infect animals, but does effectively transfer genetic material to a variety of plant species. Mice immunized sub-cutaneously with crude extracts of transgenic tobacco plants showed strong immune responses and about 40 percent of the animals were protected from HPV-induced tumors. The researchers hypothesized that the low percentage of responders could be due to the low amount of E7 antigen in the plant extract vaccine dose. Only 3 to 4 micrograms per gram of fresh leaf of E7 protein were produced and the quantity administered to the mice was 20-fold lower than that known to prevent tumor growth. The production levels of E7 by targeting it to the plant secretory pathway, which is used by cells to transport material to the outside. The researchers targeted the protein to the secretory pathway by adding so called signal sequences to the protein. The signal sequences guided the protein to specific areas for accumulation, for example to the endoplasmic reticulum, where the proteins may be retained or transported out of the cell. This offered a natural way to pre-concentrate the E7 protein. The E7 production level was five-fold higher (15 micrograms of protein per gram of fresh leaf)compared to the production level in the cytoplasm.This result may be related to a positive effect on protein stability,In the endoplasmic reticulum,there are few enzymes that degrade proteins, but many so called chaperons that help proteins to fold correctly. An additional advantage of targeting E7 proteins to the secretory pathway is that the protein purification is further simplified, because the proteins might be naturally released from the roots or leaves of the plants. Mice vaccinated with these plant extracts showed a stronger immune response and a more efficacious tumor protection than mice vaccinated with plant extracts producing the E7 protein in the cytoplasm. We believe that a further enhancement of this anti-tumor immunity can be achieved by a combination of E7-containing extracts with immunostimulatory genes, or by application of two different forms of the antigen. The fact that freeze-dried N. benthamiana leaf tissues containing high amounts of E7 antigen are stable for at least one year at room temperature further improves production and storage of the vaccine. In addition, it offers the possibility to use such plant extracts as edible vaccines. Edible plant based vaccines offer a palatable oral delivery system without the costly purification processes required for injectable vaccines. However, a great disadvantage of oral vaccine delivery is that the digestive system could degrade the antigens. This might limit the use of oral vaccines against pathogens that can survive in the harsh environment of the digestive system. By using a plant system for antigen production, however, the vaccine antigens inside the plant cells are naturally protected by the cell wall Edible vaccines are now considered to be more stable and practical in the harsh environment of the gastrointestinal tract than purified vaccine antigens. Oral immunization with tobacco plants, however, could raise the problem of toxicity, since many tobacco cultivars produce high levels of toxic alkaloids. Varieties with only low alkaloid amounts, however, have been shown to be safe production and delivery systems for therapeutic vaccines. Mice fed with such tobacco leaves showed no symptoms of toxicity. A significant step toward the development of a vaccine against cancers caused by the Human Papillomavirus type 16. Since HPV16 is an enormous problem - particularly in the developing world - a vaccine against this virus needs to be highly effective, temperature-stabile and easy to administer. Furthermore, it should be possible to produce the vaccine on a large scale and cost-efficiently. Plants, as both a production and delivery system for vaccines, meet these requirements. VACCINES - PLANT EXPRESSION SYSTEM NOTES AND PROTECTIVE CAPACITY OF THE VACCINES: Enterotoxigenic E. Coli (humans)-TOBACCO - Immunogenic when administered orally Enterotoxigenic E. coli{humans}- POTATO - Immunogenic and protective when administered orally Enterotoxigenic E. Coli{humans}- MAIZE - Immunogenic and protective when administered orally Vibrio cholerae [CHOLERA] (humans) - POTATO- Immunogenic and protective when administered orally Hepatitis B virus {humans}- TOBACCO -Extracted protein is immunogenic when administered by injection. Hepatitis B virus {humans} - POTATO - Immunogenic when administered orally Hepatitis B virus {humans}- LUPIN - Immunogenic when administered orally Hepatitis B virus {humans}- LETTUCE - Immunogenic when administered orally Norwalk virus (humans) - TOBACCO - Immunogenic when administered orally Norwalk virus (humans) - POTATO - Virus-like particles form and immunogenic when administered orally
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ANALYSIS OF SODIUM LEVELS IN JUNK FOOD BY FLAME PHOTOMETER RECENT ADVANCE IN PULSATILE DRUG DELIVERY SYSTEM A REVIEW ON FAST DISSOLVING TABLET TECHNOLOGY EDIBLE VACCINE - A GREAT BOON IN MEDICINAL SCIENCE Nanoemulsion: A Versatile Drug Delivery System

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RABIES virus (humans) -TOMATO - Intact Glycoprotein

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Literature search: A. Humanized Monoclonal Antibody Produced In Transgenic Plants For Immuno-Protection Of The Vagina Against Genital Herpes: The ability to produce monoclonal antibodies (Mabs) in plants offers the opportunity for the development of an inexpensive method of mucosol immuno protection against sexually transmitted diseases. To investigate the suitability of plant expressed Mabs for vaginal preventive applications, workers had compared a humanised anti herpes simplex virus 2 (HSV-2) Mab expressed in mammalian cell culture with the same antibody expressed in soyabean. They found these Mabs to be similar in their stability in human semen and cervical mucus over 24 hr., and their efficacy for prevention of vaginal HSV-2 infection in the mouse. B. Immunogenicity In Humans Of A Recombinant Bacterial Antigen Delivered In A Transgenic Potato: Compared with vaccine delivery by injection, oral vaccines offer the hope of more convenient immunization strategies and a more practical means of implementing universal vaccination programs throughout the world. Oral vaccines act by stimulating the immune system at effector sites (lymphoid tissue) located in the gut. Genetic engineering has been used with variable success to design living and non-living systems as a means to deliver antigens to these sites and to stimulate a desired immune response. More recently, plant biotechnology techniques have been used to create plants which contain a gene derived from a human pathogen; the resultant plant tissues will accumulate an antigenic protein encoded by the foreign DNA. In pre-clinical trials, workers found that antigenic proteins produced in transgenic plants retained immunogenic properties when purified; if injected into mice the antigen caused production of protein-specific antibodies. Moreover, in some experiments, if the plant tissues were simply fed to mice, a mucosal immune response occurred. The present study was conducted as a proof of principle to determine if humans would also develop a serum and/or mucosal immune response to an antigen delivered in an uncooked foodstuff. C. Efficacy Of A Food Plant-Based Oral Cholera Toxin B Subunit Vaccine. Transgenic potatoes were engineered to synthesize a cholera toxin B subunit (CTB) pentamer with affinity for G sub (M1) - ganglioside. Both serum and intenstinal CTB-specific antibodies were induced in orally immunized mice. Mucosal antibody titers declined gradually after the last immunization but were restored following an oral booster of transgenic potato. The cytopathic effect of cholera holotoxin (CT) on Vero cells was neutralized by serum from mice immunized with transgenic potato tissues. Following intraileal injection with CT, the plant-immunized mice showed up to a 60% reduction in diarrheal fluid accumulation in the small intestine. Protection against CT was based on inhibition of enterotoxin binding to the cell-surface receptor G sub (M1) - ganglioside. These results demonstrate the ability of transgenic food plants to generate protective immunity in mice against a bacterial enterotoxin. D. Expression Of The Rabies Virus Glycoprotein In Transgenic Tomatoes. Researchers have engineered tomato plants (Lycopersicon esculentum Mill var. UC82b) to express a gene for the glycoprotein (G-protein), which coats the outer surface the rabies virus. The recombinant constructs contained the G-protein gene from the ERA strain of rabies virus, including the signal peptide, under the control of the 35S promoter of cauliflower mosaic virus. Plants were transformed by Agrobacterium tumefaciens-medicated transformation of cotyledons and tissue culture on selective media. PCR confirmed the presence of the G-protein gene in plants surviving selection. Northern blot analysis indicated that RNA of the appropriate molecular weight was produced in both leaves and fruit of the transgenic plants. The recombinant G-protein was immunoprecipitated and detected by Western blot from leaves and fruit using different antisera. The G-protein expressed in tomato appeared as two distinct bands with apparent molecular mass of 62 and 60 kDa as compared to the 66 kDa observed for G-protein from virus grown in BHK cells. Electron microscopy of leaf tissue using immunogold-labeling and antisera specific for rabies G-protein showed localization of the G-protein to the Golgi bodies, vesicles, plasmalemma and cell walls of vascular parenchyma cells. In light of previous demonstration that orally administered rabies G-protein from the same ERA strain elicits protective immunity in animals, these transgenic plants should provide a valuable tool for the development of edible oral vaccines.
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