Abstract
Despite the development of advanced vaccine technology, as many as 15 million deaths occur annually as a result of inadequate prophylactic or therapeutic treatments against infectious diseases (World Health Organization 2008). The emphasis on profitability by the pharmaceutical industry has led to development of high-cost vaccines targeting diseases with high profit margins, resulting in an annual death toll for developing countries that is largely preventable. Daniell and co-investigators published the first expression of a vaccine antigen, cholera toxin subunit B, through transgenic tobacco chloroplasts in 2001. The polyploidy nature of the chloroplast genome enables engineering of a high copy number of target gene, while the translational machinery of the plastid directs the synthesis of bioactive proteins with proper folding, disulfide bond formation, and lipidation. Furthermore, gene integration is site-specific, expression is polycistronic, and natural gene containment occurs due to the maternal inheritance of the chloroplast genome. The chloroplast transformation technology (CTT) has been used to express proteins of bacterial, viral, protozoan, and recently mammalian origins that may be subsequently utilized in immunization strategies to produce protective or therapeutic immunity. Such chloroplast-derived vaccines represent an inexpensive and effective means of producing antigen-subunit vaccines. Furthermore, transformation of edible crops such as lettuce could generate stable orally-deliverable vaccine antigens through inexpensive field production and agricultural drying techniques. This platform could therefore obviate cold-chain logistics and the requirement of sterile injectables, drastically reducing downstream production costs of such biopharmaceuticals. With these proof-of-concept studies, focus within CTT is shifting towards establishing a viable platform for human immunization by demonstrating the functionality of such chloroplast derived proteins, emphasizing the need to develop edible plant-based alternatives to tobacco, and improving efficacy through additional peptide fusion-domains.
These authors made equal contributions.
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Aide P, Bassat Q, Alonso PL (2007) Towards an effective malaria vaccine. Arch Dis Child 92:476–479
Apel W, Schulze WX, Bock R (2010) Identification of protein stability determinants in chloroplasts. Plant J 63:636–650
Arlen PA, Falconer R, Cherukumilli S, Cole A, Cole AM, Oishi KK, Daniell H (2007) Field production and functional evaluation of chloroplast-derived interferon-alpha2b. Plant Biotechnol J 5:511–525
Arlen PA, Singleton M, Adamovicz JJ, Ding Y, Davoodi-Semiromi A, Daniell H (2008) Effective plague vaccination via oral delivery of plant cells expressing F1-V antigens in chloroplasts. Infect Immun 76:3640–3650
Bally J, Nadai M, Vitel M, Rolland A, Dumain R, Dubald M (2009) Plant physiological adaptations to the massive foreign protein synthesis occurring in recombinant chloroplasts. Plant Physiol 150:1474–1481
Boyhan D, Daniell H (2011) Low-cost production of proinsulin in tobacco and lettuce chloroplasts for injectable or oral delivery of functional insulin and C-peptide. Plant Biotechnol J 9:585–598
Chebolu S, Daniell H (2007) Stable expression of Gal/GalNAc lectin of Entamoeba histolytica in transgenic chloroplasts and immunogenicity in mice towards vaccine development for amoebiasis. Plant Biotechnol J 5:230–239
Cybulski RJ Jr, Sanz P, O’Brien AD (2009) Anthrax vaccination strategies. Mol Aspects Med 30:490–502
Daniell H (2007) Transgene containment by maternal inheritance: effective or elusive? Proc Natl Acad Sci USA 104:6879–6880
Daniell H, Lee SB, Panchal T, Wiebe PO (2001) Expression of the native cholera toxin B subunit gene and assembly as functional oligomers in transgenic tobacco chloroplasts. J Mol Biol 311:1001–1009
Daniell H, Ruiz G, Denes B, Sandberg L, Langridge W (2009a) Optimization of codon composition and regulatory elements for expression of human insulin like growth factor-1 in transgenic chloroplasts and evaluation of structural identity and function. BMC Biotechnol 9:33
Daniell H, Singh ND, Mason H, Streatfield SJ (2009b) Plant-made vaccine antigens and biopharmaceuticals. Trends Plant Sci 14:669–679
Davoodi-Semiromi A, Schreiber M, Nalapalli S, Verma D, Singh ND, Banks RK, Chakrabarti D, Daniell H (2010) Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery. Plant Biotechnol J 8:223–242
DiMichele DM (2006) Inhibitor treatment in haemophilias A and B: inhibitor diagnosis. Haemophilia 12(Suppl 6):37–41, discussion 41–32
Dreesen IA, Charpin-El Hamri G, Fussenegger M (2010) Heat-stable oral alga-based vaccine protects mice from Staphylococcus aureus infection. J Biotechnol 145:273–280
Eibl C, Zou Z, Beck A, Kim M, Mullet J, Koop HU (1999) In vivo analysis of plastid psbA, rbcL and rpl32 UTR elements by chloroplast transformation: tobacco plastid gene expression is controlled by modulation of transcript levels and translation efficiency. Plant J 19:333–345
Fernandez-San Millan A, Ortigosa SM, Hervas-Stubbs S, Corral-Martinez P, Segui-Simarro JM, Gaetan J, Coursaget P, Veramendi J (2008) Human papillomavirus L1 protein expressed in tobacco chloroplasts self-assembles into virus-like particles that are highly immunogenic. Plant Biotechnol J 6:427–441
Forman S, Le Gall F, Belton D, Evans B, Francois JL, Murray G, Sheesley D, Vandersmissen A, Yoshimura S (2009) Moving towards the global control of foot and mouth disease: an opportunity for donors. Rev Sci Tech 28:883–896
Geyer M, Peterlin BM (2001) Domain assembly, surface accessibility and sequence conservation in full length HIV-1 Nef. FEBS Lett 496:91–95
Glenz K, Bouchon B, Stehle T, Wallich R, Simon MM, Warzecha H (2006) Production of a recombinant bacterial lipoprotein in higher plant chloroplasts. Nat Biotechnol 24:76–77
Gonzalez-Rabade N, McGowan EG, Zhou F, McCabe MS, Bock R, Dix PJ, Gray JC, Ma JK (2011) Immunogenicity of chloroplast-derived HIV-1 p24 and a p24-Nef fusion protein following subcutaneous and oral administration in mice. Plant Biotechnol J. doi:10.1111/ j.1467-7652.2011.00609.x
Gorantala J, Grover S, Goel D, Rahi A, Jayadev Magani SK, Chandra S, Bhatnagar R (2011) A plant based protective antigen [PA(dIV)] vaccine expressed in chloroplasts demonstrates protective immunity in mice against anthrax. Vaccine. doi:10.1016/j.vaccine.2011.03.082
Goronzy JJ, Weyand CM (2007) The innate and adaptive immune systems. In: Goldman L, Ausiello D (eds) Cecil medicine, 23rd edn. Saunders Elsevier, Philadelphia, chap 42
Greenwood BM, Bojang K, Whitty CJ, Targett GA (2005) Malaria. Lancet 365:1487–1498
He DM, Qian KX, Shen GF, Zhang ZF, Li YN, Su ZL, Shao HB (2007) Recombination and expression of classical swine fever virus (CSFV) structural protein E2 gene in Chlamydomonas reinhardtii chroloplasts. Colloids Surf B Biointerfaces 55:26–30
Herman RA, Korjagin VA, Schafer BW (2005) Quantitative measurement of protein digestion in simulated gastric fluid. Regul Toxicol Pharmacol 41:175–184
Herman RA, Storer NP, Gao Y (2006) Digestion assays in allergenicity assessment of transgenic proteins. Environ Health Perspect 114:1154–1157
Hills CE, Brunskill NJ (2009) Cellular and physiological effects of C-peptide. Clin Sci (Lond) 116:565–574
Kanagaraj AP, Verma D, Daniell H (2011) Expression of dengue-3 premembrane and envelope polyprotein in lettuce chloroplasts. Plant Mol Biol 76:323–333
Kang TJ, Loc NH, Jang MO, Jang YS, Kim YS, Seo JE, Yang MS (2003) Expression of the B subunit of E. coli heat-labile enterotoxin in the chloroplasts of plants and its characterization. Transgenic Res 12:683–691
Kang TJ, Han SC, Kim MY, Kim YS, Yang MS (2004) Expression of non-toxic mutant of Escherichia coli heat-labile enterotoxin in tobacco chloroplasts. Protein Expr Purif 38:123–128
Kirk DD, Webb SR (2005) The next 15 years: taking plant-made vaccines beyond proof of concept. Immunol Cell Biol 83:248–256
Koya V, Moayeri M, Leppla SH, Daniell H (2005) Plant-based vaccine: mice immunized with chloroplast-derived anthrax protective antigen survive anthrax lethal toxin challenge. Infect Immun 73:8266–8274
Langeveld JP, Casal JI, Osterhaus AD, Cortes E, de Swart R, Vela C, Dalsgaard K, Puijk WC, Schaaper WM, Meloen RH (1994) First peptide vaccine providing protection against viral infection in the target animal: studies of canine parvovirus in dogs. J Virol 68:4506–4513
Lee MY, Zhou Y, Lung RW, Chye ML, Yip WK, Zee SY, Lam E (2006) Expression of viral capsid protein antigen against Epstein-Barr virus in plastids of Nicotiana tabacum cv. SR1. Biotechnol Bioeng 94:1129–1137
Lee SB, Li B, Jin S, Daniell H (2010) Expression and characterization of antimicrobial peptides Retrocyclin-101 and Protegrin-1 in chloroplasts to control viral and bacterial infections. Plant Biotechnol J 9:100–115
Lentz EM, Segretin ME, Morgenfeld MM, Wirth SA, Dus Santos MJ, Mozgovoj MV, Wigdorovitz A, Bravo-Almonacid FF (2009) High expression level of a foot and mouth disease virus epitope in tobacco transplastomic plants. Planta 231:387–395
Limaye A, Koya V, Samsam M, Daniell H (2006) Receptor-mediated oral delivery of a bioencapsulated green fluorescent protein expressed in transgenic chloroplasts into the mouse circulatory system. FASEB J 20:959–961
Loemba H, Brenner B, Parniak MA, Ma’ayan S, Spira B, Moisi D, Oliveira M, Detorio M, Wainberg MA (2002) Genetic divergence of human immunodeficiency virus type 1 Ethiopian clade C reverse transcriptase (RT) and rapid development of resistance against nonnucleoside inhibitors of RT. Antimicrob Agents Chemother 46:2087–2094
Lossl AG, Waheed MT (2011) Chloroplast-derived vaccines against human diseases: achievements, challenges and scopes. Plant Biotechnol J 9:527–539
Lotter H, Khajawa F, Stanley SL Jr, Tannich E (2000) Protection of gerbils from amebic liver abscess by vaccination with a 25-mer peptide derived from the cysteine-rich region of Entamoeba histolytica galactose-specific adherence lectin. Infect Immun 68:4416–4421
Lowy FD (1998) Staphylococcus aureus infections. N Engl J Med 339:520–532
Lowy FD (2003) Antimicrobial resistance: the example of Staphylococcus aureus. J Clin Invest 111:1265–1273
Ma JK, Drake PM, Christou P (2003) The production of recombinant pharmaceutical proteins in plants. Nat Rev Genet 4:794–805
Madesis P, Osathanunkul M, Georgopoulou U, Gisby MF, Mudd EA, Nianiou I, Tsitoura P, Mavromara P, Tsaftaris A, Day A (2009) A hepatitis C virus core polypeptide expressed in chloroplasts detects anti-core antibodies in infected human sera. J Biotechnol 145:377–386
Maher B (2008) Malaria: the end of the beginning. Nature 451:1042–1046
Matsui K, Gromowski GD, Li L, Barrett AD (2010) Characterization of a dengue type-specific epitope on dengue 3 virus envelope protein domain III. J Gen Virol 91:2249–2253
Molina A, Hervas-Stubbs S, Daniell H, Mingo-Castel AM, Veramendi J (2004) High-yield expression of a viral peptide animal vaccine in transgenic tobacco chloroplasts. Plant Biotechnol J 2:141–153
Molina A, Veramendi J, Hervas-Stubbs S (2005) Induction of neutralizing antibodies by a tobacco chloroplast-derived vaccine based on a B cell epitope from canine parvovirus. Virology 342:266–275
Morgenfeld M, Segretin ME, Wirth S, Lentz E, Zelada A, Mentaberry A, Gissmann L, Bravo-Almonacid F (2009) Potato virus X coat protein fusion to human papillomavirus 16 E7 oncoprotein enhance antigen stability and accumulation in tobacco chloroplast. Mol Biotechnol 43:243–249
Nagata M, Santamaria P, Kawamura T, Utsugi T, Yoon JW (1994) Evidence for the role of CD8+ cytotoxic T cells in the destruction of pancreatic beta-cells in nonobese diabetic mice. J Immunol 152:2042–2050
Oey M, Lohse M, Kreikemeyer B, Bock R (2009a) Exhaustion of the chloroplast protein synthesis capacity by massive expression of a highly stable protein antibiotic. Plant J 57:436–445
Oey M, Lohse M, Scharff LB, Kreikemeyer B, Bock R (2009b) Plastid production of protein antibiotics against pneumonia via a new strategy for high-level expression of antimicrobial proteins. Proc Natl Acad Sci USA 106:6579–6584
Ortigosa SM, Fernandez-San Millan A, Veramendi J (2009) Stable production of peptide antigens in transgenic tobacco chloroplasts by fusion to the p53 tetramerisation domain. Transgenic Res 19:703–709
Passalacqua KD, Bergman NH (2006) Bacillus anthracis: interactions with the host and establishment of inhalational anthrax. Future Microbiol 1:397–415
Patti JM, Hook M (1994) Microbial adhesins recognizing extracellular matrix macromolecules. Curr Opin Cell Biol 6:752–758
Pohle T, Domschke W (2003) Gastric function measurements in drug development. Br J Clin Pharmacol 56:156–164
Que YA, Moreillon P (2004) Infective endocarditis. Nat Rev Cardiol 8:322–336
Rasala BA, Muto M, Lee PA, Jager M, Cardoso RM, Behnke CA, Kirk P, Hokanson CA, Crea R, Mendez M, Mayfield SP (2010) Production of therapeutic proteins in algae, analysis of expression of seven human proteins in the chloroplast of Chlamydomonas reinhardtii. Plant Biotechnol J 8:719–733
Rigano MM, Manna C, Giulini A, Pedrazzini E, Capobianchi M, Castilletti C, Di Caro A, Ippolito G, Beggio P, De Giuli MC, Monti L, Vitale A, Cardi T (2009) Transgenic chloroplasts are efficient sites for high-yield production of the vaccinia virus envelope protein A27L in plant cellsdagger. Plant Biotechnol J 7:577–591
Rosales-Mendoza S, Alpuche-Solis AG, Soria-Guerra RE, Moreno-Fierros L, Martinez-Gonzalez L, Herrera-Diaz A, Korban SS (2009) Expression of an Escherichia coli antigenic fusion protein comprising the heat labile toxin B subunit and the heat stable toxin, and its assembly as a functional oligomer in transplastomic tobacco plants. Plant J 57:45–54
Ruhlman T, Ahangari R, Devine A, Samsam M, Daniell H (2007) Expression of cholera toxin B-proinsulin fusion protein in lettuce and tobacco chloroplasts–oral administration protects against development of insulitis in non-obese diabetic mice. Plant Biotechnol J 5:495–510
Ruhlman T, Verma D, Samson N, Daniell H (2010) The role of heterologous chloroplast sequence elements in transgene integration and expression. Plant Physiol 152:2088–2104
Rweyemamu MM, Astudillo VM (2002) Global perspective for foot and mouth disease control. Rev Sci Tech 21:765–773
Scotti N, Alagna F, Ferraiolo E, Formisano G, Sannino L, Buonaguro L, De Stradis A, Vitale A, Monti L, Grillo S, Buonaguro FM, Cardi T (2009) High-level expression of the HIV-1 Pr55gag polyprotein in transgenic tobacco chloroplasts. Planta 229:1109–1122
Sim J-S, Pak H-K, Kim D-S, Lee S-B, Kim Y-H, Hahn B-S (2009) Expression and characterization of synthetic heat-labile enterotoxin B subunit and hemagglutinin–neuraminidase-neutralizing epitope fusion protein in Escherichia coli and tobacco chloroplasts. Plant Mol Biol Rep 27:388–399
Soria-Guerra RE, Alpuche-Solis AG, Rosales-Mendoza S, Moreno-Fierros L, Bendik EM, Martinez-Gonzalez L, Korban SS (2009) Expression of a multi-epitope DPT fusion protein in transplastomic tobacco plants retains both antigenicity and immunogenicity of all three components of the functional oligomer. Planta 229:1293–1302
Staub JM, Garcia B, Graves J, Hajdukiewicz PT, Hunter P, Nehra N, Paradkar V, Schlittler M, Carroll JA, Spatola L, Ward D, Ye G, Russell DA (2000) High-yield production of a human therapeutic protein in tobacco chloroplasts. Nat Biotechnol 18:333–338
Streatfield SJ (2005) Mucosal immunization using recombinant plant-based oral vaccines. Methods Mucosal Immun 38:150–157
Thoren KL, Krantz BA (2011) The unfolding story of anthrax toxin translocation. Mol Microbiol 80:588–595
Tran M, Zhou B, Pettersson PL, Gonzalez MJ, Mayfield SP (2009) Synthesis and assembly of a full-length human monoclonal antibody in algal chloroplasts. Biotechnol Bioeng 104:663–673
Tregoning JS, Nixon P, Kuroda H, Svab Z, Clare S, Bowe F, Fairweather N, Ytterberg J, van Wijk KJ, Dougan G, Maliga P (2003) Expression of tetanus toxin Fragment C in tobacco chloroplasts. Nucleic Acids Res 31:1174–1179
Tregoning JS, Clare S, Bowe F, Edwards L, Fairweather N, Qazi O, Nixon PJ, Maliga P, Dougan G, Hussell T (2005) Protection against tetanus toxin using a plant-based vaccine. Eur J Immunol 35:1320–1326
Verhounig A, Karcher D, Bock R (2010) Inducible gene expression from the plastid genome by a synthetic riboswitch. Proc Natl Acad Sci USA 107:6204–6209
Verma D, Daniell H (2007) Chloroplast vector systems for biotechnology applications. Plant Physiol 145:1129–1143
Verma D, Kanagaraj A, Jin S, Singh ND, Kolattukudy PE, Daniell H (2010a) Chloroplast-derived enzyme cocktails hydrolyse lignocellulosic biomass and release fermentable sugars. Plant Biotechnol J 8:332–350
Verma D, Moghimi B, LoDuca PA, Singh HD, Hoffman BE, Herzog RW, Daniell H (2010b) Oral delivery of bioencapsulated coagulation factor IX prevents inhibitor formation and fatal anaphylaxis in hemophilia B mice. Proc Natl Acad Sci USA 107:7101–7106
Vines RR, Ramakrishnan G, Rogers JB, Lockhart LA, Mann BJ, Petri WA Jr (1998) Regulation of adherence and virulence by the Entamoeba histolytica lectin cytoplasmic domain, which contains a beta2 integrin motif. Mol Biol Cell 9:2069–2079
Waheed MT, Thones N, Muller M, Hassan SW, Razavi NM, Lossl E, Kaul HP, Lossl AG (2010) Transplastomic expression of a modified human papillomavirus L1 protein leading to the assembly of capsomeres in tobacco: a step towards cost-effective second-generation vaccines. Transgenic Res 20:271–282
Walker BD, Burton DR (2008) Toward an AIDS vaccine. Science 320:760–764
Wang X, Brandsma M, Tremblay R, Maxwell D, Jevnikar AM, Huner N, Ma S (2008) A novel expression platform for the production of diabetes-associated autoantigen human glutamic acid decarboxylase (hGAD65). BMC Biotechnol 8:87
Watson J, Koya V, Leppla SH, Daniell H (2004) Expression of Bacillus anthracis protective antigen in transgenic chloroplasts of tobacco, a non-food/feed crop. Vaccine 22:4374–4384
Weiner HL, da Cunha AP, Quintana F, Wu H (2011) Oral tolerance. Immunol Rev 241:241–259
Worlds Health Organization (2008) Global Burden of Diseae (GBD). The global burden of diseases 2004 update. World Health Organization, Geneva. Available from http://www.who.int/healthinfo/global_burden_disease/2004_report_update/en/index.html
Young KR, McBurney SP, Karkhanis LU, Ross TM (2006) Virus-like particles: designing an effective AIDS vaccine. Methods 40:98–117
Zhou YX, Lee MY, Ng JM, Chye ML, Yip WK, Zee SY, Lam E (2006) A truncated hepatitis E virus ORF2 protein expressed in tobacco plastids is immunogenic in mice. World J Gastroenterol 12:306–312
Zhou F, Badillo-Corona JA, Karcher D, Gonzalez-Rabade N, Piepenburg K, Borchers AM, Maloney AP, Kavanagh TA, Gray JC, Bock R (2008) High-level expression of human immunodeficiency virus antigens from the tobacco and tomato plastid genomes. Plant Biotechnol J 6:897–913
Acknowledgement
The contribution here was supported in part by funding from the Arnold and Mabel Beckman Foundation to James S. New, and by grants from USDA-CSREES 2009-39200-19972, USDA-NIFA 2010-39200-21704 and NIH R01 GM 63879–08 to Henry Daniell.
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New, J.S., Westerveld, D., Daniell, H. (2012). Chloroplast-Derived Therapeutic and Prophylactic Vaccines. In: Wang, A., Ma, S. (eds) Molecular Farming in Plants: Recent Advances and Future Prospects. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-2217-0_4
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