Transgenic plants for animal health

Edible vaccine against piglet ETEC diarrhea

J. J. Joensuu, I. Van Molle, F. Verdonck, M. Kotiaho, L. Buts, A. Ehrström, M. Peltola, H. Siljander-Rasi, Anna-Maria Nuutila, Kirsi-Marja Oksman-Caldentey, T. H. Teeri, J. Bouckaert, L. Wyns, H. De Greve, S. Panjikar, E. Cox, B. M. Goddeeris, V. Niklander-Teeri

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

Abstract

F4 fimbriae are the major colonization factors associated with porcine neonatal and postweaning diarrhea caused by enterotoxigenic Escherichia coli (ETEC). Via the chaperone/usher pathway, the F4 are assembled as long polymers of the major subunit FaeG, which also possesses the adhesive properties of the fimbriae. Being highly stable and mucosally immunogenic F4/FaeG offers a unique model system to study oral vaccination against ETEC-induced postweaning diarrhea (PWD). PWD is a major problem in piggeries worldwide and results in significant economic losses. No vaccine is currently available to protect piglets against PWD. Transgenic plants provide an economically feasible platform for large-scale production of vaccine antigens for animal health. Here, the capacity of transgenic plants to produce FaeG was evaluated. Using the model plant tobacco, FaeG was directed to different subcellular compartments by specific targeting signals. Targeting of FaeG into chloroplasts offered a superior accumulation level of 1% of total soluble proteins over the endoplasmic reticulum and the apoplast. Intrinsically, the incomplete fold of fimbrial subunits renders them unstable and susceptible to aggregation and/or proteolytical degradation in the absence of a specific periplasmic chaperone. The chloroplast-targeted FaeG was purified from tobacco and crystallized. The crystal structure shows that chloroplasts circumvent the absence of the fimbrial assembly machinery by assembling FaeG into strand-swapped dimers. Moreover, the FaeG-dimers retained the key properties of an oral vaccine, i.e. stability in gastrointestinal conditions, binding to porcine intestinal F4 receptors, and inhibition of the F4+ ETEC attachment to F4R. To investigate the oral immunogenicity, the FaeG protein was expressed in the crop plants alfalfa and barley. Desiccated alfalfa plants and barley grains stored FaeG in a stable form for years. When the transgenic alfalfa plants and cholera toxin were orally co-administered to weaned piglets, F4-specific systemic and mucosal immune responses were induced and the duration and number of F4+ E. coli excretion following F4+ ETEC challenge were reduced. In conclusion, these results suggest that transgenic plants producing the FaeG subunit protein could be used for production and delivery of oral vaccines against porcine PWD.
Original languageEnglish
Title of host publicationPlants for Human Health in the Post-Genome Era
Subtitle of host publicationPSE Congress
Place of PublicationEspoo
PublisherVTT Technical Research Centre of Finland
Pages52-53
ISBN (Electronic)978-951-38-6322-7
ISBN (Print)978-951-38-6321-0
Publication statusPublished - 2007
MoE publication typeNot Eligible
EventPSE Congress: Plants for Human Health in the Post-Genome Era - Helsinki, Finland
Duration: 26 Aug 200729 Aug 2007

Publication series

NameVTT Symposium
PublisherVTT
Number249
ISSN (Print)0357-9387
ISSN (Electronic)1445-0873

Conference

ConferencePSE Congress: Plants for Human Health in the Post-Genome Era
CountryFinland
CityHelsinki
Period26/08/0729/08/07

Fingerprint

edible vaccines
enterotoxigenic Escherichia coli
animal health
piglets
transgenic plants
diarrhea
vaccines
alfalfa
mouth
chloroplasts
fimbriae
swine
tobacco
barley
mucosal immunity
oral vaccination
cholera toxin
phytotoxins
stored grain
apoplast

Cite this

Joensuu, J. J., Van Molle, I., Verdonck, F., Kotiaho, M., Buts, L., Ehrström, A., ... Niklander-Teeri, V. (2007). Transgenic plants for animal health: Edible vaccine against piglet ETEC diarrhea. In Plants for Human Health in the Post-Genome Era: PSE Congress (pp. 52-53). [S4] Espoo: VTT Technical Research Centre of Finland. VTT Symposium, No. 249
Joensuu, J. J. ; Van Molle, I. ; Verdonck, F. ; Kotiaho, M. ; Buts, L. ; Ehrström, A. ; Peltola, M. ; Siljander-Rasi, H. ; Nuutila, Anna-Maria ; Oksman-Caldentey, Kirsi-Marja ; Teeri, T. H. ; Bouckaert, J. ; Wyns, L. ; De Greve, H. ; Panjikar, S. ; Cox, E. ; Goddeeris, B. M. ; Niklander-Teeri, V. / Transgenic plants for animal health : Edible vaccine against piglet ETEC diarrhea. Plants for Human Health in the Post-Genome Era: PSE Congress. Espoo : VTT Technical Research Centre of Finland, 2007. pp. 52-53 (VTT Symposium; No. 249).
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title = "Transgenic plants for animal health: Edible vaccine against piglet ETEC diarrhea",
abstract = "F4 fimbriae are the major colonization factors associated with porcine neonatal and postweaning diarrhea caused by enterotoxigenic Escherichia coli (ETEC). Via the chaperone/usher pathway, the F4 are assembled as long polymers of the major subunit FaeG, which also possesses the adhesive properties of the fimbriae. Being highly stable and mucosally immunogenic F4/FaeG offers a unique model system to study oral vaccination against ETEC-induced postweaning diarrhea (PWD). PWD is a major problem in piggeries worldwide and results in significant economic losses. No vaccine is currently available to protect piglets against PWD. Transgenic plants provide an economically feasible platform for large-scale production of vaccine antigens for animal health. Here, the capacity of transgenic plants to produce FaeG was evaluated. Using the model plant tobacco, FaeG was directed to different subcellular compartments by specific targeting signals. Targeting of FaeG into chloroplasts offered a superior accumulation level of 1{\%} of total soluble proteins over the endoplasmic reticulum and the apoplast. Intrinsically, the incomplete fold of fimbrial subunits renders them unstable and susceptible to aggregation and/or proteolytical degradation in the absence of a specific periplasmic chaperone. The chloroplast-targeted FaeG was purified from tobacco and crystallized. The crystal structure shows that chloroplasts circumvent the absence of the fimbrial assembly machinery by assembling FaeG into strand-swapped dimers. Moreover, the FaeG-dimers retained the key properties of an oral vaccine, i.e. stability in gastrointestinal conditions, binding to porcine intestinal F4 receptors, and inhibition of the F4+ ETEC attachment to F4R. To investigate the oral immunogenicity, the FaeG protein was expressed in the crop plants alfalfa and barley. Desiccated alfalfa plants and barley grains stored FaeG in a stable form for years. When the transgenic alfalfa plants and cholera toxin were orally co-administered to weaned piglets, F4-specific systemic and mucosal immune responses were induced and the duration and number of F4+ E. coli excretion following F4+ ETEC challenge were reduced. In conclusion, these results suggest that transgenic plants producing the FaeG subunit protein could be used for production and delivery of oral vaccines against porcine PWD.",
author = "Joensuu, {J. J.} and {Van Molle}, I. and F. Verdonck and M. Kotiaho and L. Buts and A. Ehrstr{\"o}m and M. Peltola and H. Siljander-Rasi and Anna-Maria Nuutila and Kirsi-Marja Oksman-Caldentey and Teeri, {T. H.} and J. Bouckaert and L. Wyns and {De Greve}, H. and S. Panjikar and E. Cox and Goddeeris, {B. M.} and V. Niklander-Teeri",
year = "2007",
language = "English",
isbn = "978-951-38-6321-0",
series = "VTT Symposium",
publisher = "VTT Technical Research Centre of Finland",
number = "249",
pages = "52--53",
booktitle = "Plants for Human Health in the Post-Genome Era",
address = "Finland",

}

Joensuu, JJ, Van Molle, I, Verdonck, F, Kotiaho, M, Buts, L, Ehrström, A, Peltola, M, Siljander-Rasi, H, Nuutila, A-M, Oksman-Caldentey, K-M, Teeri, TH, Bouckaert, J, Wyns, L, De Greve, H, Panjikar, S, Cox, E, Goddeeris, BM & Niklander-Teeri, V 2007, Transgenic plants for animal health: Edible vaccine against piglet ETEC diarrhea. in Plants for Human Health in the Post-Genome Era: PSE Congress., S4, VTT Technical Research Centre of Finland, Espoo, VTT Symposium, no. 249, pp. 52-53, PSE Congress: Plants for Human Health in the Post-Genome Era, Helsinki, Finland, 26/08/07.

Transgenic plants for animal health : Edible vaccine against piglet ETEC diarrhea. / Joensuu, J. J.; Van Molle, I.; Verdonck, F.; Kotiaho, M.; Buts, L.; Ehrström, A.; Peltola, M.; Siljander-Rasi, H.; Nuutila, Anna-Maria; Oksman-Caldentey, Kirsi-Marja; Teeri, T. H.; Bouckaert, J.; Wyns, L.; De Greve, H.; Panjikar, S.; Cox, E.; Goddeeris, B. M.; Niklander-Teeri, V.

Plants for Human Health in the Post-Genome Era: PSE Congress. Espoo : VTT Technical Research Centre of Finland, 2007. p. 52-53 S4 (VTT Symposium; No. 249).

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

TY - CHAP

T1 - Transgenic plants for animal health

T2 - Edible vaccine against piglet ETEC diarrhea

AU - Joensuu, J. J.

AU - Van Molle, I.

AU - Verdonck, F.

AU - Kotiaho, M.

AU - Buts, L.

AU - Ehrström, A.

AU - Peltola, M.

AU - Siljander-Rasi, H.

AU - Nuutila, Anna-Maria

AU - Oksman-Caldentey, Kirsi-Marja

AU - Teeri, T. H.

AU - Bouckaert, J.

AU - Wyns, L.

AU - De Greve, H.

AU - Panjikar, S.

AU - Cox, E.

AU - Goddeeris, B. M.

AU - Niklander-Teeri, V.

PY - 2007

Y1 - 2007

N2 - F4 fimbriae are the major colonization factors associated with porcine neonatal and postweaning diarrhea caused by enterotoxigenic Escherichia coli (ETEC). Via the chaperone/usher pathway, the F4 are assembled as long polymers of the major subunit FaeG, which also possesses the adhesive properties of the fimbriae. Being highly stable and mucosally immunogenic F4/FaeG offers a unique model system to study oral vaccination against ETEC-induced postweaning diarrhea (PWD). PWD is a major problem in piggeries worldwide and results in significant economic losses. No vaccine is currently available to protect piglets against PWD. Transgenic plants provide an economically feasible platform for large-scale production of vaccine antigens for animal health. Here, the capacity of transgenic plants to produce FaeG was evaluated. Using the model plant tobacco, FaeG was directed to different subcellular compartments by specific targeting signals. Targeting of FaeG into chloroplasts offered a superior accumulation level of 1% of total soluble proteins over the endoplasmic reticulum and the apoplast. Intrinsically, the incomplete fold of fimbrial subunits renders them unstable and susceptible to aggregation and/or proteolytical degradation in the absence of a specific periplasmic chaperone. The chloroplast-targeted FaeG was purified from tobacco and crystallized. The crystal structure shows that chloroplasts circumvent the absence of the fimbrial assembly machinery by assembling FaeG into strand-swapped dimers. Moreover, the FaeG-dimers retained the key properties of an oral vaccine, i.e. stability in gastrointestinal conditions, binding to porcine intestinal F4 receptors, and inhibition of the F4+ ETEC attachment to F4R. To investigate the oral immunogenicity, the FaeG protein was expressed in the crop plants alfalfa and barley. Desiccated alfalfa plants and barley grains stored FaeG in a stable form for years. When the transgenic alfalfa plants and cholera toxin were orally co-administered to weaned piglets, F4-specific systemic and mucosal immune responses were induced and the duration and number of F4+ E. coli excretion following F4+ ETEC challenge were reduced. In conclusion, these results suggest that transgenic plants producing the FaeG subunit protein could be used for production and delivery of oral vaccines against porcine PWD.

AB - F4 fimbriae are the major colonization factors associated with porcine neonatal and postweaning diarrhea caused by enterotoxigenic Escherichia coli (ETEC). Via the chaperone/usher pathway, the F4 are assembled as long polymers of the major subunit FaeG, which also possesses the adhesive properties of the fimbriae. Being highly stable and mucosally immunogenic F4/FaeG offers a unique model system to study oral vaccination against ETEC-induced postweaning diarrhea (PWD). PWD is a major problem in piggeries worldwide and results in significant economic losses. No vaccine is currently available to protect piglets against PWD. Transgenic plants provide an economically feasible platform for large-scale production of vaccine antigens for animal health. Here, the capacity of transgenic plants to produce FaeG was evaluated. Using the model plant tobacco, FaeG was directed to different subcellular compartments by specific targeting signals. Targeting of FaeG into chloroplasts offered a superior accumulation level of 1% of total soluble proteins over the endoplasmic reticulum and the apoplast. Intrinsically, the incomplete fold of fimbrial subunits renders them unstable and susceptible to aggregation and/or proteolytical degradation in the absence of a specific periplasmic chaperone. The chloroplast-targeted FaeG was purified from tobacco and crystallized. The crystal structure shows that chloroplasts circumvent the absence of the fimbrial assembly machinery by assembling FaeG into strand-swapped dimers. Moreover, the FaeG-dimers retained the key properties of an oral vaccine, i.e. stability in gastrointestinal conditions, binding to porcine intestinal F4 receptors, and inhibition of the F4+ ETEC attachment to F4R. To investigate the oral immunogenicity, the FaeG protein was expressed in the crop plants alfalfa and barley. Desiccated alfalfa plants and barley grains stored FaeG in a stable form for years. When the transgenic alfalfa plants and cholera toxin were orally co-administered to weaned piglets, F4-specific systemic and mucosal immune responses were induced and the duration and number of F4+ E. coli excretion following F4+ ETEC challenge were reduced. In conclusion, these results suggest that transgenic plants producing the FaeG subunit protein could be used for production and delivery of oral vaccines against porcine PWD.

M3 - Conference abstract in proceedings

SN - 978-951-38-6321-0

T3 - VTT Symposium

SP - 52

EP - 53

BT - Plants for Human Health in the Post-Genome Era

PB - VTT Technical Research Centre of Finland

CY - Espoo

ER -

Joensuu JJ, Van Molle I, Verdonck F, Kotiaho M, Buts L, Ehrström A et al. Transgenic plants for animal health: Edible vaccine against piglet ETEC diarrhea. In Plants for Human Health in the Post-Genome Era: PSE Congress. Espoo: VTT Technical Research Centre of Finland. 2007. p. 52-53. S4. (VTT Symposium; No. 249).