Cholera toxin B (CTB) is functional as an adjuvant for cytoplasmatic proteins if directed to the endoplasmatic reticulum (ER), but not to the cytoplasm of plants

Mikschofsky, Heike; König, Patricia; Keil, Günther M.; Hammer, Martin; Schirrmeier, Horst; Broer, Inge

doi:10.1016/j.plantsci.2009.03.010

Cited by 19 publications

(17 citation statements)

References 44 publications

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“…Synthetic sequences encoding CTB and VP60 using the codon preference of tobacco (Mikschofsky et al 2009b) were used. Transgenic potatoes transformed with pGK-ctb SEKDEL , pGK-vp60 SEKDEL and pGK were characterised by PCR, RT-PCR and Northern blot analyses, using the total RNA from leaves with vp60-specific primers and a ctb gene probe (Mikschofsky et al 2009b, c. The events with the highest expression levels, Desiree-VP60-6 and -17 and Albatros-CTB-204 and -218, were selected (Mikschofsky et al 2009b). The nearisogenic varieties (NIV), Desiree and Albatros, derived from laboratory cultures and the transgenic events, Desiree-NPTII-6 and Albatros-NPTII-205, carrying only the marker gene served as controls.…”

Section: Resultsmentioning

confidence: 99%

“…Desiree and Albatros, were cultivated in vitro in growth chambers at 22°C/18°C with a 16 h/8 h light/dark cycle on Murashige and Skoog (MS) medium (Murashige and Skoog 1962), containing 20 g/l of sucrose. The binary vectors pGK-ctb SEKDEL , pGK-vp60 SEKDEL and pGK (Mikschofsky et al 2009b, c were introduced into Agrobacterium tumefaciens LBA4404 (Hoekema et al 1983). The Solanum tubersosum cv.…”

Section: Methodsmentioning

confidence: 99%

“…Because there are no in vitro cell culture systems available to propagate RHDV, all of the commercial VP60-based vaccines are prepared from the livers of experimentally infected rabbits. Recently, the successful expression of VP60 in tobacco (Fernandez-Fernandez et al 2001;Mikschofsky et al 2009b), carrot (Mikschofsky et al 2009a, c;pea (Mikschofsky et al 2009d) and potato (Castanon et al 1999;Castanon et al 2002) have been reported.…”

Section: Introductionmentioning

confidence: 99%

“…We have introduced a synthetic vp60 gene (Mikschofsky et al 2009c) into the potato variety Desiree and a synthetic ctb gene into the variety Albatros (Mikschofsky et al 2009b) and selected two events per construct for comprehensive expression analysis under greenhouse and field conditions.…”

Section: Introductionmentioning

confidence: 99%

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Greenhouse and field cultivations of antigen-expressing potatoes focusing on the variability in plant constituents and antigen expression

Mikschofsky

Heilmann

Schmidtke³

et al. 2011

Plant Mol Biol

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The production of plant-derived pharmaceuticals essentially requires stable concentrations of plant constituents, especially recombinant proteins; nonetheless, soil and seasonal variations might drastically interfere with this stability. In addition, variability might depend on the plant organ used for production. Therefore, we investigated the variability in plant constituents and antigen expression in potato plants under greenhouse and field growth conditions and in leaves compared to tubers. Using potatoes expressing VP60, the only structural capsid protein of the rabbit haemorrhagic disease virus (RHDV), CTB, the non-toxic B subunit (CTB) of the cholera toxin (CTA-CTB(5)) and the marker protein NPTII (neomycinphosphotransferase) as a model, we compare greenhouse and field production of potato-derived antigens. The influence of the production organ turned out to be transgene specific. In general, yield, plant quality and transgene expression levels in the field were higher than or similar to those observed in the greenhouse. The variation (CV) of major plant constituents and the amount of transgene-encoded protein was not influenced by the higher variation of soil properties observed in the field. Amazingly, for specific events, the variability in the model protein concentrations was often lower under field than under greenhouse conditions. The changes in gene expression under environmental stress conditions in the field observed in another event do not reduce the positive influence on variability since events like these should excluded from production. Hence, it can be concluded that for specific applications, field production of transgenic plants producing pharmaceuticals is superior to greenhouse production, even concerning the stability of transgene expression over different years. On the basis of our results, we expect equal or even higher expression levels with lower variability of recombinant pharmaceuticals in the field compared to greenhouse production combined with approximately 10 times higher tuber yield in the field.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Greenhouse and field cultivations of antigen-expressing potatoes focusing on the variability in plant constituents and antigen expression

Mikschofsky

Heilmann

Schmidtke³

et al. 2011

Plant Mol Biol

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“…The pentameric B subunit of the cholera toxin, CT-B, is structurally and functionally very similar to LT-B and has thus been the main target for developing plant-produced candidate vaccines against cholera as well as traveler's diarrhea. CT-B has been expressed in potato (Arakawa et al, 1997;He et al, 2007;Mikschofsky et al, 2009), tobacco (Daniell et al, 2001Kang et al, 2004b, Kang et al, 2006cMishra et al, 2006;Mikschofsky et al, 2009;Tiwari et al, 2009;Rattanapisit et al, 2013), carrot (Kim et al, 2009), tomato leaves and fruit (Meena et al, 2002;Jiang et al, 2007;Loc et al, 2011b), lettuce (Huy et al, 2011), rice (Soh et al, 2015) and maize seeds (Karaman et al, 2012). As it is a potential mucosal and parenteral adjuvant and an effective carrier for chemically or genetically linked antigens, CT-B has often been expressed in the form of fusion proteins with antigens which do not induce sufficient immune response when orally applied on their own, much like LT-B.…”

Section: Cholera Toxin Subunit Bmentioning

confidence: 99%

Production of vaccines for treatment of infectious diseases by transgenic plants

Ledl

Luthar

2016

AAS

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Since the first pathogen antigen was expressed in transgenic plants with the aim of producing edible vaccine in early 1990s, transgenic plants have become a well-established expression system for production of alternative vaccines against various human and animal infectious diseases. The main focus of plant expression systems in the last five years has been on improving expression of well-studied antigens such as porcine reproductive and respiratory syndrome (PRRSV), bovine viral diarrhea disease virus (BVDV), footh and mouth disease virus (FMDV), hepatitis B surface antigen (HBsAg), rabies G protein, rotavirus, Newcastle disease virus (NDV), Norwalk virus capsid protein (NVCP), avian influenza virus H5N1, Escherichia coli heat-labile enterotoxin subunit B (LT-B), cholera toxin B (CT-B), human immunodeficiency virus (HIV), artherosclerosis, ebola and anthrax. Significant increases in expression have been obtained using improved expression vectors, different plant species and transformation methods.Key words: transgenic plants; vaccine production; recombinant proteins; antibodies; infectious diseases; risk assessment IZVLEČEK ZDRAVLJENJE NALEZLJIVIH BOLEZNI S CEPIVI PRIDOBLJENIMI S TRANSGENIMI RASTLINAMIPridobivanje aktivnih sestavin za cepiva s transgenimi rastlinami se je začelo v devetdesetih letih prejšnjega stoletja. Od takrat se transgene rastline uveljavljajo kot alternativni ekspresijski sistem za sintezo aktivnih komponent za cepiva proti številnim humanim in živalskim nalezljivim boleznim. V zadnjih petih letih je bil glavni poudarek razvoja namenjen optimizaciji rastlinskih ekspresijskih sistemov za dobro proučene antigene, kot so virus prašičjega respiratornega in reproduktivnega sindroma, virus goveje virusne diareje, virus parkljevke, površinski antigen hepatitisa B, G protein virusa stekline, rotavirus, virus atipične kokošje kuge, plaščni protein Norwalk virusa, sev ptičje gripe H5N1, temperaturno nestabilna B podenota enterotoksina Escherischia coli, kolera toksin B, HIV, arterioskleroza, ebola in antraks. Uporaba optimiziranih transformacijskih metod in ekspresijskih vektorjev v kombinaciji s primerno izbrano rastlinsko vrsto je v večini naštetih primerov vodila do opaznega izboljšanja sinteze posameznih učinkovin.

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Genetic Engineering of Vegetable Crops

Sidhu

Chellan

2010

Handbook of Vegetables and Vegetable Processing

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Cholera toxin B (CTB) is functional as an adjuvant for cytoplasmatic proteins if directed to the endoplasmatic reticulum (ER), but not to the cytoplasm of plants

Cited by 19 publications

References 44 publications

Greenhouse and field cultivations of antigen-expressing potatoes focusing on the variability in plant constituents and antigen expression

Greenhouse and field cultivations of antigen-expressing potatoes focusing on the variability in plant constituents and antigen expression

Production of vaccines for treatment of infectious diseases by transgenic plants

Genetic Engineering of Vegetable Crops

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