edible vaccines
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Seminar titleEdible vaccines
Masume jahedM.Sc student of Agricultural
Biotechnology
Shahid beheshti university
Guided by Dr. hossein askari
Molecular farming
In putIsolation of gene of interest
Expression hosts for molecular farming
BacteriaYeast Mamalian cell cultureTransgenic Plants plant cells culture
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(Spök et al., 2008)
Out put
Vaccines antigens Therapeutics products Nutritional components Industrial products Bio plastics
Plant Molecular farming
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(Sharma ansd Sharma 2009)
What is 'molecular farming in plants'?
Advantages : Plant as Bioreactor
Significantly lower production costs than with transgenic animals, fermentation or bioreactors
Plants do not contain known human pathogens (such as virions, etc.) That could contaminate the final product.
(Sahu et al., 2014)4
low cost of production low cost of production
High safty High safty
ability to fold and assemble eucaryote complex proteins accurately ability to fold and assemble eucaryote complex proteins accurately
Rapid scalability Rapid scalability
Tyes of vaccines
Different types of currently available vaccines
Conventional vaccine Live attenuated vaccines Killed vaccines purified subunit vaccines (inactivated toxin)
Costly Longer time is needed for it to be produced Heat sensitive Prone to microbial contamination
recombinant subunit vaccineAdvances in molecular biology techniques during the 1980s, helped in the development of new strategies for the production of subunit vaccines.
Vaccines produced by biotechnological method are stable at room temperature, unlike traditional vaccine which needs cold chain storage which increases the yearly cost to preserve.
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History of Edible vaccines
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In 1990, Streptococcus mutans surface protein A was expressed in transgenic tobacco and given to mice. learning of a World Health Organization call for inexpensive oral vaccines that needed no refrigeration.
(Langridge, 2000)
In the early 1990s Charles J. Arntzen, then at Texas A&M University, conceived of a way to solve many of the problems that bar vaccines from reaching all too many children in developing nations.
History of Ecdible vaccines
Mason et al 1992 Haq et al 1995 McGarvey et al 1995 Mason et al 1996 Hein et al 1996 ….
Hepatitis
Hepatitis B surface antigen (HBsAg)
Tobacco/leaf
Norwalk virus (NV)
Gastroenteritis Norwalk virus capsid protein (NVCP)
Potato/tubertobacco/leaf
Rabies virus
Rabies
Rabies virus glycoprotein(RVG)
Tomato/leaf, fruit
V. Cholerae
Cholera
Cholera toxin B subunit (CTB)
Tobacco/leaf
E. Coli
Diarrhea
Heat labile toxin B subunit (LTB)
Potato/tuber, tobacco/leaf
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Production of Edible vaccine
1) stable nuclear transformation of a crop species that are grown in the field or a greenhouse
2) Stable plastid transformation of a crop species
3) transient transformation of a crop species
4) stable transformation of a plant species that is grown hydroponically or in in vitro systems
stable nuclear transformation of a crop species that are grown in the field or a greenhouse
Molecular analysis of T0 transgenic plants
expression cassette
Molecular analysis of T1,
T2, … generations of
transgenic plants
Putative transgenic plants
9transgenic plants
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Stable plastid transformation of a crop species
Advantages
Chloroplast system, represents the highest recombinant protein accumulation achieved so far in plants. plastids are inherited through maternal tissues in most species and the pollen does not contain chloroplasts, hence the transgene may not be transferable, thereby allaying public concerns.
Disadvantages
It is also envisaged that protein stability will change over time even with refrigeration.
(Meyers et al., 2010; Horn et al., 2004)
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transient transformation of a crop species
(Thomas et al., 2002)
Stable transformation of a plant species that is grown hydroponically or in in vitro systems
agroinfiltration (Agrobacterium-mediated delivery)
genetically modified plant viral vectors
Clinical trials
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Protein purification
Oral administration
Injected routes
(Daniell et al., 2009)
Application of Edible vaccines
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1) infectious diseaseAIDSMeaslesSmallpoxTransmissible gastroenteritis TuberculosisInfuenzaAnthraxTetanusSevere Acute Respiratory Syndrome (Sars)Human PapillomavirusCysticercosisPlagueFoot and mouth diseaseCentral nervous system diseaseNewcastle diseaseRabbit hemorrhagic syndromesCanine parvovirus diseaseBluetongue
Preventing :2) Autoimmune disease
Type-I Diabetes multiple sclerosis
3) Cancer therapy Colon cancer Cervical cancer
(Tiwari et al., 2009; Hefferon, 2010; Sharma et al., 2004)
Features of different plant host systems
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(Saxena and Shweta Rawat, 2014)
Advantages Disadvantages
Tomato Relatively efficient transformation system fruit is edible raw
Relatively low fruit protein content; acidic fruit may be incompatible with some antigens or for delivery to
infants
Tobaco Facile and efficient transformation system
Toxic alkaloids incompatible with oral delivery
Poatato Industrial tuber processing well established
unpalatablein raw form; cooking might cause denaturation
and poor immunogenicity of vaccine
Banana Cultivated widely in developing countries where vaccines are needed
Inefficient transformation system
Advantages of Edible vaccines
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do not require administration by injection do not require separation and purification of vaccines from plant materials necessary syringe & needles not required Low cost Eliminate trained medical personnel
Enhance compliance in children contamination risk is low as sterile conditions are not and Induction of both mucosul and systemic immunization
Induction of mucosul immunization
16(Mishra et al., 2008)
Plant cell protect the antigent against enzymatic attack
o Plant cell degradation by digestive/ bacterial enzyme in intestineo Antigens released near the Intestinal lining over Peyer’s patcheso Stimulate follicles develop germinal centreo Taken up by M cellso Penetrates the intestinal epitheliumo Accumulates within organized lymphoid structureo M cells express class II MHCo transported across the mucous membrane by M cellso Activate B cells within lymphoid follicleso Migration to MALTo Differentiate into Plasma cells Ig A production in plasma cells and memory cells.oTransporte to Lumeno Interacte with antigeno Neutralize the infectious agent
limitation, Concerns and Safety Aspects of Edible Vaccine
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Selection of best plant is difficult Some food cannot be eaten raw (e.g. potato) and needs cooking which will denature or weaken the protein present in it. Consistency of dosage form fruit to fruit, plant-to-plant, and generation-to-generation is not similar Dosage of vaccines would be variable Possibility of development of immunotolerance to the vaccine protein or peptide. Standardization of dosage in case of edible vaccine, is very important. Possible allergic reaction to plant componant is yet another remote concern.
From Seeds, freeze-dried fruits and leaves: powder form vaccines are produced. Recent Development : “Edible Vaccines” now more popular as “Plant Vaccines”
Not convenient for infants.
The potential gene flow to weeds or related crops through pollination or seed contamination there are issues about PDMs accidentally entering the food chain and being consumed by non-target organisms.
Conclusions
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www.who.int/ Global Immunization Data
an estimated 21.8 million infants worldwide are still not being reached by routine immunization services. Inadequate supply of vaccines, lack of access to health workers, and insufficient political and financial support account for a large proportion of people who start but don’t finish national immunization schedules.
future prospects
There are several technical and logistic problems which need to be addressed before plant derived edible vaccine becomes a reality )Tiwari et al., 2009).
The optimal dosage required to produce immunity and duration of the immune response Distribution and marketing issue Examining issues related to commercialization Issues relating to the ethical, social, biosafety and environmental impact.
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References
1- Floss, D. M., Falkenburg, D., & Conrad, U. (2007). Production of vaccines and therapeutic antibodies for veterinary applications in transgenic plants: an overview. Transgenic Research, 16(3), 315-332. 2- Sahu, P. K., Patel, T. S., Sahu, P., Singh, S., Tirkey, P., & Sharma, D. (2014). Molecular Farming : A biotechnological approach in agriculture for production of useful metabolites. International Journal of Research in Biotechnology and Biochemistry, 4(2): 23-30.3- Daniell, H., Singh, N. D., Mason, H., & Streatfield, S. J. (2009). Plant-made vaccine antigens and biopharmaceuticals. Trends in plant science, 14(12), 669-679.4- Lal, P., Ramachandran, V. G., Goyal, R., & Sharma, R. (2007). Edible vaccines: current status and future. Indian journal of medical microbiology, 25(2), 93.5- Tiwari, S., Verma, P. C., Singh, P. K., & Tuli, R. (2009). Plants as bioreactors for the production of vaccine antigens. Biotechnology advances, 27(4), 449-467.6- Mason, H. S., Lam, D. M., & Arntzen, C. J. (1992). Expression of hepatitis B surface antigen in transgenic plants. Proceedings of the National Academy of Sciences, 89(24), 11745-11749.7- Streatfield, S. J., & Howard, J. A. (2003). Plant-based vaccines. International journal for parasitology, 33(5), 479-493.
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References
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8- Saxena, J., & Rawat, S. (2014). Edible Vaccines. In Advances in Biotechnology (pp. 207-226). Springer India.9- Shah, C. P., Trivedi, M. N., Vachhani, U. D., & Joshi, V. J. (1990). Edible Vaccine: A Better Way for Immunization. pharmaceuticals, 4, 5.10- Sabalza, M., Christou, P., & Capell, T. (2014). Recombinant plant-derived pharmaceutical proteins: current technical and economic bottlenecks. Biotechnology letters, 36(12), 2367-2379.11- Obembe, O. O., Popoola, J. O., Leelavathi, S., & Reddy, S. V. (2011). Advances in plant molecular farming. Biotechnology advances, 29(2), 210-222.12- Naithani, M., Viswanath, D., & Urs, P. (2014). Edible vaccines-a review. International Journal of Pharmacotherapy, 4(1), 58-61.13- Fahad, S., Khan, F. A., Pandupuspitasari, N. S., Ahmed, M. M., Liao, Y. C., Waheed, M. T., ... & Huang, J. (2014). Recent developments in therapeutic protein expression technologies in plants. Biotechnology letters, 1-15.14- Spök, A., Karner, S., Stein, A. J., & Rodríguez-Cerezo, E. (2008). Plant molecular farming. Opportunities and challenges. JRC Scientific and Technical Reports.
References
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15- Hefferon, K. (2010). Clinical trials fuel the promise of plant-derived vaccines. Am J Clin Med, 7, 30-7.16- Langridge, William HR. (2000). "Edible vaccines.“ Scientific american-american edition- 283.3 : 48-53.17- Sharma, A. K., & Sharma, M. K. (2009). Plants as bioreactors: Recent developments and emerging opportunities. Biotechnology advances, 27(6), 811-832.
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