Expression of Cholera Toxin Subunits in Plantsa

Hein, Mich B.; Yeo, T. C.; Wang, Fei; Sturtevant, Ann

doi:10.1111/j.1749-6632.1996.tb32490.x

Cited by 21 publications

(6 citation statements)

References 19 publications

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“…The expressed CTB subunit retained its native antigenicity, and induced the generation of both mucosal and systemic anti-CT antibodies at levels sufficient to generate protective immunity by oral immunization in experimental mice. But its antigenicity was lost by more than 50% after the potatoes were cooked (Hein et al 1996;Arakawa et al 1997Arakawa et al , 1998Arakawa et al , 1999. The Tomato plant is one of the selected plants for transformation as oral vaccine since its fruits are edible fresh.…”

Section: Introductionmentioning

confidence: 98%

Cholera toxin B protein in transgenic tomato fruit induces systemic immune response in mice

Jiang

Peng

et al. 2007

Transgenic Res

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Cholera toxin B (CTB) subunit is a well-characterized antigen against cholera. Transgenic plants can offer an inexpensive and safe source of edible CTB vaccine and may be one of the best candidates for the production of plant vaccines. The present study aimed to develop transgenic tomato expressing CTB protein, especially in the ripening tomato fruit under the control of the tomato fruit-specific E8 promoter by using Agrobacterium-mediated transformation. Transgenic plants were selected using PCR and Southern blot analysis. Exogenous protein extracted from leaf, stem, and fruit tissues of transgenic plants was detected by ELISA and Western blot analysis, showing specific expression in the ripening fruit, with the highest amount of CTB protein being 0.081% of total soluble protein. Gavage of mice with ripe transgenic tomato fruits induced both serum and mucosal CTB specific antibodies. These results demonstrate the immunogenicity of the CTB protein in transgenic tomato and provide a considerable basis for exploring the utilization of CTB in the development of tomato-based edible vaccine against cholera. The rCTB antigen resulted in much lower antibody titers than an equal amount of exogenous CTB in transgenic fruits, suggesting the protective effect of the fibrous tissue of the fruit to the exogenous CTB protein against the degradation of protease in the digestive tracts of mice.

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Section: Introductionmentioning

confidence: 98%

Cholera toxin B protein in transgenic tomato fruit induces systemic immune response in mice

Jiang

Peng

et al. 2007

Transgenic Res

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“…Previous studies have shown that transgenic plant material expressing hepatitis B surface antigen can induce a primary immune response that, upon parenteral boosting, can generate a long-lasting antibody response (Thanawala et al 1995). Cholera toxin B subunit (CTB) has been expressed in transgenic tobacco (Hein et al 1996), potato (Arakawa et al 1997), and tomato (Jani et al 2002). Potato-expressed CTB has been found to induce a protective immune response in mice (Arakawa et al 1998).…”

Section: Introductionmentioning

confidence: 98%

Studies on the immunogenic potential of plant-expressed cholera toxin B subunit

et al. 2004

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Nicotiana tabacum var. Samsun was transformed via Agrobacterium-mediated transformation with a gene encoding the cholera toxin B subunit (CTB) of Vibrio cholerae, modified to contain a sequence coding for an endoplasmic reticulum retention signal (SEKDEL), under the control of the cauliflower mosaic virus 35S promoter. Total protein from the transgenic leaf tissue was isolated and an aliquot containing 5 microg recombinant CTB was injected intradermally into Balb/c (H2K(d)) mice. CTB-specific serum IgG was detected in animals that had been administered plant-expressed or native purified CTB. A T-cell proliferation study using splenocytes and cytokine estimations in supernatants generated by in vitro stimulation of macrophages isolated from the immuno-primed animals was carried out. Inhibition of proliferation of T lymphocytes was observed in splenic T lymphocytes isolated from animals injected with either native or plant-expressed CTB. Macrophages isolated from mice immunised with native or plant-expressed CTB showed enhanced secretion of interleukin-10 but secretion of lipopolysaccharide-induced interleukin-12 and tumor necrosis factor alpha was inhibited. These studies suggest that plant-expressed protein behaved like native CTB with regards to effects on T-cell proliferation and cytokine levels, indicating the suitability of plant expression systems for the production of bacterial antigens, which could be used as edible vaccine. The transgene was found to be inherited in the progeny and was expressed to yield a pentameric form of CTB as evident by its interaction with G(M1) ganglioside.

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“…Expression of proteins from a variety of microbial and viral pathogens showed the feasibility of transgenic plant expression systems for the production of subunit vaccines. The proteins expressed include those from the Hepatitis B surface antigen (HbsAg) in tobacco and lettuce (Ehsani, Khabiri and Domansky, 1997;Mason, Lam and Arntzen, 1992), a rabies virus antigen in tomato (McGarvey et al, 1995), a cholera antigen in tobacco and potato (Arakawa et al, 1997;Hein et al, 1996), a recombinant bacterial antigen (LT-B) in tobacco and potato (Haq et al, 1995), the Norwalk virus capsid protein (NVCP) in tobacco and potato (Mason et al, 1996) and a human cytomegalovirus antigen in tobacco (Tackaberry et al, 1999).…”

Section: Recombinant Subunit Vaccinesmentioning

confidence: 99%