Two decades of plant-based candidate vaccines: A review of the chimeric protein approaches

Plant Cell Reports

Volumn 30, Issue 8, 2011, Pages 1367-1382

  Soria Guerra, Ruth Elena ^a   Moreno Fierros, Leticia ^b   Rosales Mendoza, Sergio ^a  

^a UNIVERSIDAD AUTÓNOMA DE SAN LUIS POTOSÍ   (Mexico)

^b UNIVERSIDAD NACIONAL AUTÓNOMA DE MÉXICO   (Mexico)

Author keywords

Broad immune protection; Fusion protein; Molecular farming; Oral immunization; Plant based vaccine

Indexed keywords

EID: 85027952290     PISSN: 07217714     EISSN: None     Source Type: Journal    
DOI: 10.1007/s00299-011-1065-3     Document Type: Review

References (105)

1. Alvarez ML, Cardineau GA (2010) Prevention of bubonic and pneumonic plague using plant derived vaccines. Biotechnol Adv 28: 184-196.
2. Alvarez ML, Pinyerd HL, Crisantes JD, Rigano M, Pinkhasov J, Walmsley AM, Mason HS, Cardineau GA (2006) Plant-made subunit vaccine against pneumonic and bubonic plague is orally immunogenic in mice. Vaccine 24: 2477-2490.
3. Anderson GW, Leary SE, Williamson DE, Titball RW, Welkos SL, Worsham PL, Friedlander AM (1996) Recombinant V antigen protects mice against pneumonic and bubonic plague caused by F1-capsule-positive and -negative strains of Yersinia pestis. Infect Immun 64: 4580-4585.
4. Andrews GP, Heath DG, Anderson GW, Welkos SL, Friedlander AM (1996) Fraction 1 capsular antigen purification from Yersinia pestis CO92 and from an E. coli recombinant strain and efficacy against lethal plague challenge. Infect Immun 64: 2180-2187.
5. Andrianov V, Brodzik R, Spitsin S, Bandurska K, McManus H, Koprowski H, Golovkin M (2010) Production of recombinant anthrax toxin receptor (ATR/CMG2) fused with human Fc in planta. Prot Expr Pur 70: 158-162.
6. 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-Vs in chloroplasts. Infect Immun 76: 3640-3650.
7. Ashkenazi A, Chamow S (1997) Immunoadhesins as research tools and therapeutic agents. Curr Opin Immunol 9: 195-200.
8. Bandurska K, Brodzik R, Spitsin S, Kohl T, Portocarrero C, Smirnov Y, Pogrebnyak N, Sirko A, Koprowski H, Golovkin M (2008) Plant- produced hepatitis B core protein chimera carrying anthrax protective antigen domain-4. Hybridoma 27: 241-247.
9. Berzofsky JA, Ahlers JD, Belyakov IM (2001) Strategies for designing and optimizing new generation vaccines. Nat Rev Immunol 1: 209-219.
10. Boothe J, Nykiforuk C, Shen Y, Zaplachinski S, Szarka S, Kuhlman P, Murray E, Morck D, Moloney MM (2010) Seed-based expression systems for plant molecular farming. Plant Biotechnol J 8: 588-606.
11. Cardenas L, Clements JD (1993) Development of mucosal protection against the heat stable enterotoxin (ST) of Escherichia coli by oral immunization with a genetic fusion delivered by a bacterial vector. Infect Immun 61: 4629-4636.
12. Cardi T, Lenzi P, Maliga P (2010) Chloroplasts as expression platforms for plant-produced vaccines. Expert Rev Vaccines 9: 893-911.
13. CDC (1992) Pertussis vaccination: acellular pertussis vaccine for the fourth and fifth doses of the DTP series update to supplementary ACIP statement. Recommendations of the Advisory Committee on Immunization Practices (ACIP). MMWR 41 (No. RR-15): 1-5.
14. Chichester JA, Musiychuk K, Farrance CE, Mett V, Lyons J, Mett V, Yusibov V (2009) A single component two-valent LcrV-F1 vaccine protects non-human primates against pneumonic plague. Vaccine 27: 3471-3474.
15. Choi NW, Estes MK, Langridge WH (2005) Synthesis and assembly of a cholera toxin B subunit-rotavirus VP7 fusion protein in transgenic potato. Mol Biotechnol 31: 193-202.
16. Coffman RL, Sher A, Seder RA (2010) Vaccine adjuvants: putting innate immunity to work. Immunity. doi: 10. 1016/j. immuni. 2010. 10. 002.
17. Conley AJ, Zhu H, Le LC, Jevnikar AM, Lee BH, Brandle JE, Menassa R (2010) Recombinant protein production in a variety of Nicotiana hosts: a comparative analysis. Plant Biotechnol J. doi: 10. 1111/j. 1467-7652. 2010. 00563. x.
18. Conrad U, Plagmann I, Malchow S, Sack M, Floss DM, Kruglov AA, Nedospasov SA, Rose-John S, Scheller J (2011) ELPylated anti-human TNF therapeutic single-domain antibodies for prevention of lethal septic shock. Plant Biotechnol J 9: 22-31.
19. Crasto CJ, Feng J (2000) LINKER: a program to generate linker sequences for fusion proteins. Protein Eng 13: 309-312.
20. Crompton DW (2001) Ascaris and ascariasis. Adv Parasitol 48: 285-375.
21. 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.
22. Estrada A, Li B, Laarveld B (1998) Adjuvant action of Chenopodium quinoa saponins on the induction of antibody responses to intragastric and intranasal administered antigens in mice. Comp Immunol Microbiol Infect Dis 21: 225-236.
23. Field M (1979) Modes of action of enterotoxins from Vibrio cholerae and Escherichia coli. Rev Infect Dis 1: 918-926.
24. Floss DM, Schallau K, Rose-John S, Conrad U, Scheller J (2009) Elastin-like polypeptides revolutionize recombinant protein expression and their biomedical application. Trends Biotechnol 28: 37-45.
25. Floss DM, Mockey M, Zanello G, Brosson D, Diogon M, Frutos R, Bruel T, Rodrigues V, Garzon E, Chevaleyre C, Berri M, Salmon H, Conrad U, Dedieu L (2010) Expression and immunogenicity of the mycobacterial Ag85B/ESAT-6 antigens produced in transgenic plants by elastin-like peptide fusion strategy. J Biomed Biotechnol 2010: 274346.
26. Gleba Y, Klimyuk V, Marillonnet S (2005) Magnifection-a new platform for expressing recombinant vaccines in plants. Vaccine 23: 2042-2048.
27. Gleba Y, Klimyuk V, Marillonnet S (2007) Viral vectors for the expression of proteins in plants. Curr Opin Biotechnol 18: 134-141.
28. Greco R, Michel M, Guetard D, Cervantes-Gonzalez M, Pelucchi N, Wain-Hobson S, Sala F, Sala M (2007) Production of recombinant HIV-1/HBV virus-like particles in Nicotiana tabacum and Arabidopsis thaliana plants for a bivalent plant-based vaccine. Vaccine 25: 8228-8240.
29. Guerrero GG, Moreno-Fierros L (2007) Carrier potential properties of Bacillus thuringiensis Cry1A toxins for a diphtheria toxin epitope. Scand J Immunol 66: 610-618.
30. Gutiérrez-Ortega A, Avila-Moreno F, Saucedo-Arias LJ, Sánchez-Torres C, Gómez-Lim MA (2004) Expression of a single-chain human interleukin-12 gene in transgenic tobacco plants and functional studies. Biotechnol Bioeng 85: 734-740.
31. Gutiérrez-Ortega A, Sandoval-Montes C, de Olivera-Flores TJ, Santos-Argumedo L, Gómez-Lim MA (2005) Expression of functional interleukin-12 from mouse in transgenic tomato plants. Transgenic Res 14: 877-885.
32. Hammond RW, Nemchinov LG (2009) Plant production of veterinary vaccines and therapeutics. Curr Top Microbiol Immunol 332: 79-102.
33. Haq TA, Mason HS, Clements JD, Arntzen CJ (1995) Oral immunization with a recombinant bacterial antigen produced in transgenic plants. Science 268: 714-716.
34. Howard JA (2005) Commercialization of biopharmaceutical and bioindustrial proteins from plants. Crop Sci 45: 468-472.
35. Huang Z, Elkin G, Maloney BJ, Beuhner N, Arntzen CJ, Thanavala Y, Mason HS (2005) Virus-like particle expression and assembly in plants: hepatitis B and Norwalk viruses. Vaccine 23: 1851-1858.
36. Kalthoff D, Giritch A, Geisler K, Bettmann U, Klimyuk V, Gleba Y, Hehnen HS, Beer M (2010) Immunization with plant-expressed hemagglutinin protects chickens from lethal highly pathogenic avian influenza virus H5N1 challenge infection. J Virol 84: 12002-12010.
37. Karlsson Hedestam GB, Fouchier RA, Phogat S, Burton DR, Sodroski J, Wyatt RT (2008) The challenges of eliciting neutralizing antibodies to HIV-1 and to influenza virus. Nat Rev Microbiol 6: 143-155.
38. Kim TG, Galloway DR, Langridge WH (2004a) Synthesis and assembly of anthrax lethal factor-cholera toxin B-subunit fusion protein in transgenic potato. Mol Biotechnol 28: 175-183.
39. Kim TG, Gruber A, Langridge WH (2004b) HIV-1 gp120 V3 cholera toxin B subunit fusion gene expression in transgenic potato. Protein Expr Purif 37: 196-202.
40. Kim TG, Gruber A, Ruprecht RM, Langridge WH (2004c) Synthesis and assembly of SIVmac Gag p27 capsid protein cholera toxin B subunit fusion protein in transgenic potato. Mol Biotechnol 28: 33-40.
41. Kim TG, Ruprecht R, Langridge WH (2004d) Synthesis and assembly of a cholera toxin B subunit SHIV 89. 6p Tat fusion protein in transgenic potato. Protein Expr Pur 35: 313-319.
42. Kuzel S, Vydra J, Triska J, Vrchotova N, Hruby M, Cigler P (2009) Elicitation of pharmacologically active substances in an intact medical plant. J Agric Food Chem 57: 7907-7911.
43. Lavelle EC (2005) Generation of improved mucosal vaccines by induction of innate immunity. Cell Mol Life Sci 62: 2750-2770.
44. Lee JY, Yu J, Henderson D, Langridge WH (2004) Plant-synthesized E. coli CFA/I fimbrial protein protects Caco-2 cells from bacterial attachment. Vaccine 23: 222-231.
45. Li D, O'Leary J, Huang Y, Huner NP, Jevnikar AM, Ma S (2006) Expression of cholera toxin B subunit and the B chain of human insulin as a fusion protein in transgenic tobacco plants. Plant Cell Rep 25: 417-424.
46. Licciardi PV, Underwood JR (2010) Plant-derived medicines: a novel class of immunological adjuvants. Int Immunopharmacol (in press).
47. Makare N, Bodhankar S, Rangari V (2001) Immunomodulatory activity of alcoholic extract of Mangifera indica L. in mice. J Ethnopharmacol 78: 133-137.
48. Martínez CA, Topal E, Giulietti AM, Talou JR, Mason H (2010) Exploring different strategies to express Dengue virus envelope protein in a plant system. Biotechnol Lett 32: 867-875.
49. Mason HS, Lam DM, Arntzen CJ (1992) Expression of hepatitis B surface antigen in transgenic plants. Proc Natl Acad Sci USA 89: 11745-11749.
50. Massa S, Franconi R, Brandi R, Muller A, Mett V, Yusibov V, Venuti A (2010) Anti-cancer activity of plant-produced HPV16 E7 vaccine. Vaccine 25: 3018-3021.
51. Matoba N, Kajiura H, Cherni I, Doran JD, Bomsel M, Fujiyama K, Mor TS (2009) Biochemical and immunological characterization of the plant-derived candidate human immunodeficiency virus type 1 mucosal vaccine CTB-MPR. 649-684. Plant Biotech J 7: 129-145.
52. Matsumoto Y, Suzuki S, Nozoye T, Yamakawa T, Takashima Y, Arakawa T, Tsuji N, Takaiwa F, Hayashi Y (2009) Oral immunogenicity and protective efficacy in mice of transgenic rice plants producing a vaccine candidate antigen (As16) of Ascaris suum fused with cholera toxin B subunit. Transgenic Res 18: 185-192.
53. Menassa R, Du C, Yin ZQ, Ma S, Poussier P, Brandle J, Jevnikar AM (2007) Therapeutic effectiveness of orally administered transgenic low-alkaloid tobacco expressing human interleukin-10 in a mouse model of colitis. Plant Biotechnol J 5: 50-59.
54. Mett V, Lyons J, Musiychuk K, Chichester JA, Brasil T, Couch R, Sherwood R, Palmer GA, Streatfield SJ, Yusibov V (2007) A plant-produced plague vaccine candidate confers protection to monkeys. Vaccine 25: 3014-3017.
55. Michel M, Lone YC, Centlivre M, Roux P, Wain-Hobson S, Sala M (2007) Optimisation of secretion of recombinant HBsAg virus-like particles: impact on the development of HIV-1/HBV bivalent vaccines. Vaccine 25: 1901-1911.
56. Nagai T, Yamada H (1994) In vivo anti-influenza virus activity of kampo (Japanese herbal) medicine "Sho-seiryu-to" and its mode of action. Int J Immunopharmacol 16: 605-613.
57. Nagai T, Yamada H (1998) In vivo anti-influenza virus activity of Kampo (Japanese herbal) medicine "Sho-seiryu-to"-stimulation of mucosal immune system and effect on allergic pulmonary inflammation model mice. Immunopharmacol Immunotoxicol 20: 267-281.
58. Nagai T, Urata M, Yamada H (1996) In vivo anti-influenza virus activity of Kampo (Japanese herbal) medicine "Sho-seiryu-to"-effects on aged mice, against subtypes of a viruses and B virus, and therapeutic effect. Immunopharmacol Immunotoxicol 18: 193-208.
59. Nagai T, Suzuki Y, Kiyohara H, Susa E, Kato T, Nagamine T, Hagiwara Y, Tamura S, Yabe T, Aizawa C, Yamada H (2001) Onjisaponins, from the root of Polygala tenuifolia Willdenow, as effective adjuvants for nasal influenza and diphtheria-pertussis-tetanus vaccines. Vaccine 19: 4824-4834.
60. Nagai T, Kiyohara H, Munakata K, Shirahata T, Sunazuka T, Harigaya Y, Yamada H (2002) Pinellic acid from the tuber of Pinellia ternata Breitenbach as an effective oral adjuvant for nasal influenza vaccine. Int Immunopharmacol 2: 1183-1193.
61. Nashar TO, Amin T, Marcello A, Hirst TR (1993) Current progress in the development of the B subunits of cholera toxin and Escherichia coli heat-labile enterotoxin as carriers for the oral delivery of heterologous antigens and epitopes. Vaccine 11: 235-240.
62. Nozoye T, Takaiwa F, Tsuji N, Yamakawa T, Arakawa T, Hayashi Y, Matsumoto Y (2009) Production of Ascaris suum As14 protein and its fusion protein with cholera toxin B subunit in rice seeds. J Vet Med Sci 71: 995-1000.
63. Obregon P, Chargelegue D, Drake PM, Prada A, Nuttall J, Frigerio L, Ma JK (2006) HIV-1 p24-immunoglobulin fusion molecule: a new strategy for plant-based protein production. Plant Biotech J 4: 195-207.
64. Patel J, Zhu H, Menassa R, Gyenis L, Richman A, Brandle J (2007) Elastin-like polypeptide fusions enhance the accumulation of recombinant proteins in tobacco leaves. Transgenic Res 16: 239-249.
65. Paul M, Ma JK (2010) Plant-made immunogens and effective delivery strategies. Expert Rev Vaccines 9: 821-833.
66. Pleschka S, Stein M, Schoop R, Hudson JB (2009) Anti-viral properties and mode of action of standardized Echinacea purpurea extract against highly pathogenic avian influenza virus (H5N1, H7N7) and swine-origin H1N1 (S-OIV). Virol J 6: 197.
67. Prentice MB, Rahalison L (2007) Plague. Lancet 369: 1196-1207.
68. Qian B, Shen H, Liang W, Guo X, Zhang Ch, Wang Y, Li G, Wu A, Cao K, Zhang D (2008) Immunogenicity of recombinant hepatitis B virus surface antigen fused with preS1 epitopes expressed in rice seeds. Transgenic Res 17: 621-631.
69. Quan FS, Compans RW, Cho YK, Kang SM (2007) Ginseng and Salviae herbs play a role as immune activators and modulate immune responses during influenza virus infection. Vaccine 25: 272-282.
70. Rehman J, Dillow JM, Carter SM, Chou J, Le B, Maisel AS (1999) Increased production of antigen-specific immunoglobulins G and M following in vivo treatment with the medicinal plants Echinacea angustifolia and Hydrastis canadensis. Immunol Lett 68: 391-395.
71. Rigano MM, Alvarez ML, Pinkhasov J, Jin Y, Sala F, Arntzen CJ, Walmsley AM (2004) Production of a fusion protein consisting of the enterotoxigenic Escherichia coli heat-labile toxin B subunit and a tuberculosis antigen in Arabidopsis thaliana. Plant Cell Rep 22: 502-508.
72. Rigano MM, Dreitz S, Kipnis AP, Izzo AA, Walmsley AM (2006) Oral immunogenicity of a plant-made, subunit, tuberculosis vaccine. Vaccine 24: 691-695.
73. Robinson CR, Sauer RT (1998) Optimizing the stability of single-chain proteins by linker length and composition mutagenesis. Proc Natl Acad Sci USA 95: 5929-5934.
74. Rosales-Mendoza S, Alpuche-Solís AG, Soria-Guerra RE, Moreno-Fierros L, Martínez-González L, Herrera-Díaz 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.
75. Rosales-Mendoza S, Soria-Guerra RE, Moreno-Fierros L, Martínez-González L, Alpuche-Solís AG, Korban SS (2010a) Expression of an immunogenic F1-V fusion protein in lettuce as a plant-based vaccine against plague. Planta 232: 409-416.
76. Rosales-Mendoza S, Soria-Guerra RE, Moreno-Fierros L, Han Y, Alpuche-Solís AG, Korban SS (2010b) Transgenic carrot tap roots express the immunogenic F1-V fusion protein from Yersinia pestis are immunogenic in mice. J Plant Physiol 168: 174-180.
77. Rosales-Mendoza S, Soria-Guerra RE, Moreno-Fierros L, Govea-Alonso DO, Herrera-Díaz A, Korban SS, Alpuche-Solís AG (2011) Immunogenicity of nuclear-encoded LTB: ST fusion protein from Escherichia coli expressed in tobacco plants. Plant Cell Rep. doi: 10. 1007/s00299-011-1023-0.
78. 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.
79. Ryan EJ, Daly LM, Mills KH (2001) Immunomodulators and delivery systems for vaccination by mucosal routes. Trends Biotechnol 19: 293-304.
80. Sánchez-Hernández C, Gutiérrez-Ortega A, Aguilar-León D, Hernández-Pando R, Gómez-Lim M, Gómez-García B (2010) In vivo activity of plant-based interleukin-12 in the lung of Balb/c mouse. BMC Res Notes 3: 151.
81. Santi L, Batchelor L, Huang Z, Hjelm B, Kilbourne J, Arntzen CJ, Chen Q, Mason HS (2008) An efficient plant viral expression system generating orally immunogenic Norwalk virus-like particles. Vaccine 26: 1846-1854.
82. Schaetti C (2009) Vaccines for enteric diseases: update on recent developments. Expert Rev Vaccines 8: 1653-1655.
83. Scheller J, Henggeler D, Viviani A, Conrad U (2004) Purification of spider silk-elastin from transgenic plants and application for human chondrocyte proliferation. Transgenic Res 13: 51-57.
84. Sharma MK, Jani D, Thungapathra M, Gautam JK, Meena LS, Singh Y, Ghosh A, Tyagi AK, Sharma AK (2008a) Expression of accessory colonization factor subunit A (ACFA) of Vibrio cholerae and ACFA fused to cholera toxin B subunit in transgenic tomato (Solanum lycopersicum). J Biotechnol 135: 22-27.
85. Sharma MK, Singh NK, Jani D, Sisodia R, Thungapathra M, Gautam JK, Meena LS, Singh Y, Ghosh A, Tyagi AK, Sharma AK (2008b) Expression of toxin co-regulated pilus subunit A (TCPA) of Vibrio cholerae and its immunogenic epitopes fused to cholera toxin B subunit in transgenic tomato (Solanum lycopersicum). Plant Cell Rep 27: 307-318.
86. Shchelkunov SN, Salyaev RK, Pozdnyakov SG, Rekoslavskaya NI, Nesterov AE, Ryzhova TS, Sumtsova VM, Pakova NV, Mishutina UO, Kopytina TV, Hammond RW (2006) Immunogenicity of a novel, bivalent, plant-based oral vaccine against hepatitis B and human immunodeficiency viruses. Biotechnol Lett 28: 959-967.
87. Shin EA, Lee JY, Kim TG, Park YK, Langridge WH (2006) Synthesis and assembly of an adjuvanted Porphyromonas gingivalis fimbrial antigen fusion protein in plants. Protein Expr Purif 47: 99-109.
88. Sing A, Rost D, Tvardovskaia N, Roggenkamp A, Wiedemann A, Kirschning CJ, Aepfelbacher M, Heesemann J (2002) Yersinia V-antigen exploits Toll-like receptor 2 and CD14 for interleukin 10-mediated immunosuppression. J Exp Med 196: 1017-1024.
89. Smith ML, Richter L, Arntzen CJ, Shuler ML, Mason HS (2003) Structural characterization of plant-derived hepatitis B surface antigen employed in oral immunization studies. Vaccine 21: 4011-4021.
90. Soria-Guerra RE, Rosales-Mendoza S, Márquez-Mercado C, López-Revilla L, Castillo-Collazo R, Alpuche-Solís AG (2007) Transgenic tomatoes express an antigenic polypeptide containing epitopes of the diphtheria, pertussis and tetanus exotoxins, encoded by a synthetic gene. Plant Cell Rep 26: 961-968.
91. Soria-Guerra RE, Alpuche-Solís AG, Rosales-Mendoza S, Moreno-Fierros L, Martínez-González L, Bendik E, 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.
92. Soria-Guerra RE, Rosales-Mendoza S, Moreno-Fierros L, Lopez-Revilla R, Alpuche-Solis AG (2011) Oral immunogenicity of tomato-derived sDPT polypeptide containing Corynebacterium diphtheriae, Bordetella pertussis and Clostridium tetani exotoxin epítopes. Plant Cell Rep 30: 417-424.
93. Spangler BD (1992) Structure and function of cholera toxin and the related Escherichia coli heat-labile enterotoxin. Microbiol Rev 56: 622-647.
94. Stoger E, Ma JKC, Fischer R, Christou P (2005) Sowing the seeds of success: pharmaceutical proteins from plants. Curr Opin Biotech 16: 167-173.
95. Titball RW, Leary SE (1998) Plague. British Medical Bulletin 54 pp. 625-633.
96. Vázquez RI, Moreno-Fierros L, Neri-Bazán L, De La Riva GA, López-Revilla R (1999) Bacillus thuringiensis Cry1Ac protoxin is a potent systemic and mucosal adjuvant. Scand J Immunol 49: 578-584.
97. Vázquez-Padrón RI, Moreno-Fierros L, Neri-Bazán L, de la Riva GA, López-Revilla R (1999) Intragastric and intraperitoneal administration of Cry1Ac protoxin from Bacillus thuringiensis induces systemic and mucosal antibody responses in mice. Life Sci 64: 1897-1912.
98. Walmsley AM, Alvarez ML, Jin Y, Kirk DD, Lee SM, Pinkhasov J, Rigano MM, Arntzen CJ, Mason HS (2003) Expression of the B subunit of Escherichia coli heat-labile enterotoxin as a fusion protein in transgenic tomato. Plant Cell Rep 21: 1020-1026.
99. Welkos SL, Davis KM, Pitt LM, Worsham PL, Friedlander AM (1995) Studies on the contribution of the F1-capsule-associated plasmid pFra to the virulence of Yersinia pestis. Contrib Microbiol Immunol 13: 299-305.
100. Williamson ED, Stephen ME, Griffin KF, Green M, Russel P, Leary SEC, Oyston PCF, Easterbrook T, Reddin KM, Robinson A, Titball RW (1995) A new improved sub-unit vaccine for plague: the basis of protection. FEMS Immunol Med Microbiol 12: 223-230.
101. Williamson ED, Eley SM, Stagg AJ, Green M, Russell P, Titball RW (1997) A sub-unit vaccine elicits IgG in serum, spleen cell culture and bronchial washing and protects immunized animals against pneumonic plague. Vaccine 15: 1079-1084.
102. Yu J, Langridge WH (2001) A plant-based multicomponent vaccine protects mice from enteric diseases. Nat Biotechnol 19: 548-552.
103. Zhang B, Yang YH, Lin YM, Rao Q, Zheng GG, Wu KF (2003) Expression and production of bioactive human interleukin-18 in transgenic tobacco plants. Biotechnol Lett 25: 1629-1635.
104. Zhou F, Badillo-Corona JA, Karcher D, Gonzalez-Rabade N, Piepenburg K, Borchers AM, Maloney A, Kavanagh TA, Gray JC, Bock R (2008) High-level expression of human immunodeficiency virus antigens from the tobacco and tomato plastid genomes. Plant Biotech J 6: 897-913.
105. Zimmermann J, Saalbach I, Jahn D, Giersberg M, Haehnel S, Wedel J, Macek J, Zoufal K, Glünder G, Falkenburg D, Kipriyanov SM (2009) Antibody expressing pea seeds as fodder for prevention of gastrointestinal parasitic infections in chickens. BMC Biotechnology 9: 74.