Journal Article

Mice protected by oral immunization with Lactobacillus reuteri secreting fusion protein of Escherichia coli enterotoxin subunit protein

Chi-Ming Wu and Tung-Ching Chung

in FEMS Immunology & Medical Microbiology

Published on behalf of Federation of Microbiological Societies

Volume 50, issue 3, pages 354-365
Published in print August 2007 | ISSN: 0928-8244
Published online July 2007 | e-ISSN: 1574-695X | DOI: https://dx.doi.org/10.1111/j.1574-695X.2007.00255.x
Mice protected by oral immunization with Lactobacillus reuteri secreting fusion protein of Escherichia coli enterotoxin subunit protein

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Abstract

A green fluorescent protein (gfp) gene was ligated to the Lactobacillus reuteri-specific nisin-inducible expression-secretion vector pNIES, generating a pNIES-GFP vector capable of secreting the cloned gene as a GFP-fusion protein with fluorescent activity. To develop this system as a live vehicle carrying the heat-stable enterotoxin (ST) and heat-labile enterotoxin B (LTB) of the enterotoxigenic Escherichia coli (ETEC), a recombinant 5′-ST-LTB-3′ DNA fragment was cloned into pNIES-GFP. The resulting L. reuteri/pNIES-GFP:STLTB system was found to possess the capability of adhering to the mice gut, secreting GFP:STLTB product at 0.14 and 0.026 pgcell−1 under induced and noninduced conditions, respectively. Further analysis of the GFP:STLTB product confirmed its ganglioside-binding ability, LTB antigenicity and relative freedom from the ST-associated toxicity, making it suitable for use as an oral vaccine in mice. Oral inoculation of the L. reuteri/pNIES-GFP:STLTB culture in mice elicited significant (P<0.01) serum IgG and mucosal IgA antibodies against the STLTB antigen. These immunized mice were subsequently challenged with ETEC and showed full protection against the fluid influx response in the gut. This is the first report of using L. reuteri as a vaccine carrier to induce complete immunologic protection against ETEC.

Keywords: Escherichia coli enterotoxin; Lactobacillus reuteri; oral vaccine; green fluorescent protein

Journal Article.  7373 words.  Illustrated.

Subjects: Medical Microbiology and Virology ; Biotechnology ; Genetics and Genomics ; Microbiology ; Molecular and Cell Biology

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