Diarrheal diseases caused by Vibrio cholerae and enterotoxigenic Escherichia coli (ETEC) are worldwide health problems that might be prevented with vaccines based on edible plants expressing the B subunit from either the cholera toxin (CTB) or the E. coli heat labile toxin (LTB). In this work we analyzed the immunity induced in Balb/c mice by ingestion of three weekly doses of 10 μg of LTB derived from transgenic carrot material. Although the anti-LTB serum immunoglobulin G (IgG) and intestinal IgA antibody responses were higher with 10 μg-doses of pure bacterial recombinant LTB (rLTB), the transgenic carrot material also elicited significant serum and intestinal antibody responses. Serum anti-LTB IgG1 antibodies predominated over IgG2a antibodies, suggesting that mainly Th2 responses were induced. A decrease of intestinal fluid accumulation after cholera toxin challenge was observed in mice immunized with either rLTB or LTB-containing carrot material. These results demonstrate that ingestion of carrot-derived LTB induces antitoxin systemic and intestinal immunity in mice and suggest that transgenic carrots expressing LTB may be used as an effective edible vaccine against cholera and ETEC diarrhea in humans.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Belisle BW, Twiddy EM, Holmes RK (1984) Monoclonal antibodies with an expanded repertoire of specificities and potent neutralizing activity for Escherichia coli heat-labile enterotoxin. Infect Immun 46:759–764
Black RE (1993) Epidemiology of diarrheal disease: implications for control by vaccines. Vaccine 11:100–106
Boedeker EC (2004) Vaccines for enterotoxigenic Escherichia coli: current status. Curr Opin Gastroenterol 21:15–19
Chikwamba R, McMurray J, Shou H, Frame B, Pegg SE, Scott P, Mason H, Wang K (2002a) Expression of a synthetic E. coli heat-labile enterotoxin B sub-unit (LT-B) in maize. Mol Breed 10:253–265
Chikwamba R, Cunnick J, Hathaway D, McMurray J, Mason H, Wang K (2002b) A functional antigen in a practical crop: LT-B producing maize protects mice against Escherichia coli heat labile enterotoxin (LT) and cholera toxin (CT). Transgenic Res 11:479–493
Curtis RI, Cardineau CA (1990) Oral immunization by transgenic plants. World Patent Application, WO 90/02484. (http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&u=/netahtml/search-adv.htm&r=6&p=1&f=G&l=50&d=ptxt&S1=5,686,079&OS=5,686,079&RS=5,686,079)
Daniell H, Streatfield SJ, Wycoff K (2001) Medical molecular farming: production of antibodies, biopharmaceuticals and edible vaccines in plants. Trends Plant Sci 6:219–226
Haq TA, Mason HS, Clements JD, Arntzen CJ (1995) Oral immunization with a recombinant bacterial antigen produced in transgenic plants. Science 268:714–716
Klipstein FA, Engert RF, Clements JD, Houghten RA (1984) Differences in cross-protection in rats immunized with the B subunits of cholera toxin and Escherichia coli heat-labile toxin. Infect Immun 43:811–816
Mason HS, Haq TA, Clements JD, Arntzen CJ (1998) Edible vaccine protects mice against Escherichia coli heat-labile enterotoxin (LT): potatoes expressing a synthetic LT-B gene. Vaccine 16:1336–1343
Mason H, Warzecha H, Mor T, Arntzen C (2002) Edible plant vaccines: applications for prophylactic and therapeutic molecular medicine. Trends Mol Med 8:324–329
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–479
Nataro JP, Kaper JB (1998) Diarrheagenic Escherichia coli. Clin Microbiol Rev 11:142–201
Richardson SH, Giles JC, Kruger KS (1984) Sealed adult mice: new model for enterotoxin evaluation. Infect Immun 43:482–486
Rosales-Mendoza S, Soria-Guerra RE, Olivera-Flores MTJ, López-Revilla R, Argüello-Astorga GR, Jiménez-Bremont JF, García de la Cruz RF, Loyola-Rodríguez JP, Alpuche-Solís AG (2007) Expression of Escherichia coli heat-leabile enterotoxin B subunit (LT-B) in carrot (Daucus carota L.). Plant Cell Rep 26(7):969–976 doi:10.1007/s00299-007-0310-2
Simon PW (1984) Carrot genetics. Plant Mol Biol Rep 2:54–63
Sixma TK, Pronk SE, Kalk KH, Wartna ES, van Zanten BA, Witholt B, Hol WG (1991) Crystal structure of a cholera toxin-related heat-labile enterotoxin from E. coli. Nature (London) 351:371–377
Spangler BD (1992) Structure and function of cholera toxin and related Escherichia coli heat-labile enterotoxin. Microbiol Rev 56:622–647
Streatfield SJ, Jilka JM, Hood EE, Turner DD, Bailey MR, Mayor JM, Woodard SL, Beifuss KK, Horn ME, Delaney DE, Tizard IR, Howard JA (2001) Plant-based vaccines: unique advantages. Vaccine 19:2742–2748
Tacket CO, Mason HS, Losonsky G, Clements JD, Levine MM, Arntzen CJ (1998) Immunogenicity in humans of a recombinant bacterial antigen delivered in a transgenic potato. Nat Med 4:607–609
Tacket CO, Pasetti MF, Edelman R, Howard JA, Streatfield S (2004) Immunogenicity of recombinant LT-B delivered orally to humans in transgenic corn. Vaccine 22:4385–4389
WHO. World Health Organization (2005) Global immunization. Vision and strategy 2006–2015. http://www.who.int/vaccines-documents/
Wolf MK (1997) Occurrence, distribution and associations of O and H serogroups, colonization factor antigens, and toxins of enterotoxigenic Escherichia coli. Clin Microbiol Rev 10:569–584
We wish to thank Dr. John Clements for his generous gift of pure rLTB. This research was partially funded by an UCMexUS grant to RLR as well as grant 37048-B to AAS and graduate scholarships to SRM and RSG from CONACYT (Mexico).
Communicated by P. Lakshmanan.
About this article
Cite this article
Rosales-Mendoza, S., Soria-Guerra, R.E., López-Revilla, R. et al. Ingestion of transgenic carrots expressing the Escherichia coli heat-labile enterotoxin B subunit protects mice against cholera toxin challenge. Plant Cell Rep 27, 79–84 (2008). https://doi.org/10.1007/s00299-007-0439-z
- Edible vaccine
- Protective immunity