4.4 Article

Protective Immunity Against Enterotoxigenic Escherichia coli by Oral Vaccination of Engineered Lactococcus lactis

Journal

CURRENT MICROBIOLOGY
Volume 78, Issue 9, Pages 3464-3473

Publisher

SPRINGER
DOI: 10.1007/s00284-021-02601-x

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Funding

  1. Shahed University [4/127]

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The study developed an oral vaccine based on lactic acid bacteria against ETEC, a leading cause of diarrhea in children. Experimental results showed that oral administration of the engineered L. lactis significantly increased the production of IgA in the intestine, neutralized the toxin, and reduced the pathological symptoms caused by diarrhea.
Enterotoxigenic Escherichia coli (ETEC) is one of the leading causes of diarrhea in children globally, and thus suitable vaccines are desired. Antigen display on lactic acid bacteria is a reliable approach for efficient oral vaccination and preventing bowel diseases. To develop an oral vaccine against ETEC, the gene of the binding domain from heat-labile toxin (LTB), a key ETEC virulence factor, was codon-optimized and cloned into a construct containing a signal peptide and an anchor for display on L. lactis. Bioinformatics analysis showed a codon adaptation index of 0.95 for the codon-optimized gene. Cell surface expression of LTB was confirmed by transmission electron microscopy and blotting. White New Zealand rabbits were immunized per os (PO) with the recombinant L. lactis, and the antibody titers were assayed with ELISA. In vitro neutralization assay was performed using mouse adrenal tumor cells and rabbit ileal loop test was performed as the in vivo assay. ELISA results indicated that oral administration of the engineered L. lactis elicited a significant production of IgA in the intestine. In vitro neutralization assay showed that the effect of the toxin could be neutralized with 500 mu g/ml of IgG isolated from the oral vaccine group. Furthermore, the dose of ETEC causing fluid accumulation in the ileal loop test showed a tenfold increase in rabbits immunized with either recombinant L. lactis or LTB protein compared to other groups. Our results imply that recombinant L. lactis could potentially be an effective live oral vaccine against ETEC toxicity.

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