Expression of Escherichia coli heat-labile enterotoxin B subunit in transgenic tomato (Solanum lycopersicum L.) fruit

We report a feasibility study for expressing the LTB protein (Escherichia coli heat-labile enterotoxin B subunit) via Agrobacterium-mediated transformation of tomato (Solanum lycopersicum L.). We produced five regenerated plants obtained on the selection medium supplemented with an antibiotic. Stable integrations of the LTB gene into the genome of these plants were confirmed by Southern blot hybridization. Western blot analysis showed that only two of the five T<sub>0 </sub>transgenic tomato plants expressed the pentameric LTB protein in the fruits. An enzyme-linked immunosorbent assay indicated that these two plants synthesized the LTB protein bound specifically to GM1 ganglioside, suggesting that the LTB subunits formed active pentamers. The LTB protein produced in tomatoes can be a potential candidate for inexpensive, safe, and effective plant-based vaccines.

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Functional Pentameric Formation via Coexpression of the Escherichia coli Heat-Labile Enterotoxin B Subunit and Its Fusion Protein Subunit with a Neutralizing Epitope of ApxIIA Exotoxin Improves the Mucosal Immunogenicity and Protection against Challenge by Actinobacillus pleuropneumoniae

Clinical and Vaccine Immunology ◽

10.1128/cvi.05230-11 ◽

2011 ◽

Vol 18 (12) ◽

pp. 2168-2177 ◽

Cited By ~ 7

Author(s):

Jung-Mi Kim ◽

Seung-Moon Park ◽

Jung-Ae Kim ◽

Jin-Ah Park ◽

Min-Hee Yi ◽

...

Keyword(s):

Escherichia Coli ◽

Fusion Protein ◽

Actinobacillus Pleuropneumoniae ◽

Enzyme Linked Immunosorbent Assay ◽

Recombinant Yeast ◽

Content Type ◽

Fusion Construct ◽

B Subunit ◽

Heat Labile ◽

Enterotoxin B

ABSTRACTA coexpression strategy inSaccharomyces cerevisiaeusing episomal and integrative vectors for theEscherichia coliheat-labile enterotoxin B subunit (LTB) and a fusion protein of an ApxIIA toxin epitope produced byActinobacillus pleuropneumoniaecoupled to LTB, respectively, was adapted for the hetero-oligomerization of LTB and the LTB fusion construct. Enzyme-linked immunosorbent assay (ELISA) with GM1 ganglioside indicated that the LTB fusion construct, along with LTB, was oligomerized to make the functional heteropentameric form, which can bind to receptors on the mucosal epithelium. The antigen-specific antibody titer of mice orally administered antigen was increased when using recombinant yeast coexpressing the pentameric form instead of recombinant yeast expressing either the LTB fusion form or antigen alone. Better protection against challenge infection withA. pleuropneumoniaewas also observed for coexpression in recombinant yeast compared with others. The present study clearly indicated that the coexpression strategy enabled the LTB fusion construct to participate in the pentameric formation, resulting in an improved induction of systemic and mucosal immune responses.

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Attenuated Salmonella Gallinarum secreting an Escherichia coli heat-labile enterotoxin B subunit protein as an adjuvant for oral vaccination against fowl typhoid

Veterinary Immunology and Immunopathology ◽

10.1016/j.vetimm.2012.09.006 ◽

2012 ◽

Vol 150 (3-4) ◽

pp. 149-160 ◽

Cited By ~ 9

Author(s):

Byung Woo Jeon ◽

Chetan V. Jawale ◽

Seung Hwan Kim ◽

John Hwa Lee

Keyword(s):

Escherichia Coli ◽

Oral Vaccination ◽

Salmonella Gallinarum ◽

Subunit Protein ◽

B Subunit ◽

Heat Labile ◽

Fowl Typhoid ◽

Enterotoxin B

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Tissue culture and expression of Escherichia coli heat-labile enterotoxin B subunit in transgenic Peperomia pellucida

Protein Expression and Purification ◽

10.1016/j.pep.2010.02.010 ◽

2010 ◽

Vol 72 (1) ◽

pp. 82-86 ◽

Cited By ~ 11

Author(s):

Nguyen Hoang Loc ◽

Nguyen Hoang Bach ◽

Tae-Geum Kim ◽

Moon-Sik Yang

Keyword(s):

Escherichia Coli ◽

Tissue Culture ◽

B Subunit ◽

Heat Labile ◽

Enterotoxin B

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Escherichia coli Heat-Labile Enterotoxin B Subunit Is a More Potent Mucosal Adjuvant than Its Closely Related Homologue, the B Subunit of Cholera Toxin

Infection and Immunity ◽

10.1128/iai.69.5.3476-3482.2001 ◽

2001 ◽

Vol 69 (5) ◽

pp. 3476-3482 ◽

Cited By ~ 57

Author(s):

Douglas G. Millar ◽

Timothy R. Hirst ◽

Denis P. Snider

Keyword(s):

Escherichia Coli ◽

Cholera Toxin ◽

Lymphocyte Proliferation ◽

Vaccine Adjuvants ◽

B Subunit ◽

Heat Labile ◽

Antibody Titers ◽

Enterotoxin B ◽

Draining Lymph Nodes ◽

Stimulation Of

ABSTRACT Although cholera toxin (Ctx) and Escherichia coliheat-labile enterotoxin (Etx) are known to be potent mucosal adjuvants, it remains controversial whether the adjuvanticity of the holotoxins extends to their nontoxic, receptor-binding B subunits. Here, we have systematically evaluated the comparative adjuvant properties of highly purified recombinant EtxB and CtxB. EtxB was found to be a more potent adjuvant than CtxB, stimulating responses to hen egg lysozyme when the two were coadministered to mice intranasally, as assessed by enhanced serum and secretory antibody titers as well as by stimulation of lymphocyte proliferation in spleen and draining lymph nodes. These results indicate that, although structurally very similar, EtxB and CtxB have strikingly different immunostimulatory properties and should not be considered equivalent as prospective vaccine adjuvants.

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