Characterization of a Mutant Escherichia coli Heat-Labile Toxin, LT(R192G/L211A), as a Safe and Effective Oral Adjuvant

American Society for Microbiology - Tập 18 Số 4 - Trang 546-551 - 2011
Elizabeth B. Norton1, Louise B. Lawson2, Lucy C. Freytag2, John D. Clements2
1Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, Louisiana 70112, USA
2Department of Microbiology & Immunology, Tulane University School of Medicine, New Orleans, Louisiana 70112

Tóm tắt

ABSTRACT

Despite the fact that the adjuvant properties of the heat-labile enterotoxins ofEscherichia coli(LT) andVibrio cholerae(CT) have been known for more than 20 years, there are no available oral vaccines containing these molecules as adjuvants, primarily because they are both very potent enterotoxins. A number of attempts with various degrees of success have been made to reduce or eliminate the enterotoxicity of LT and CT so they can safely be used as oral adjuvants or immunogens. In this report we characterize the structural, enzymatic, enterotoxic, and adjuvant properties of a novel mutant of LT, designated LT(R192G/L211A), or dmLT. dmLT was not sensitive to trypsin activation, had reduced enzymatic activity for induction of cyclic AMP in Caco-2 cells, and exhibited no enterotoxicity in the patent mouse assay. Importantly, dmLT retained the ability to function as an oral adjuvant for a coadministered antigen (tetanus toxoid) and to elicit anti-LT antibodies.In vitroandin vivodata suggest that the reduced enterotoxicity of this molecule compared to native LT or the single mutant, LT(R192G), is a consequence of increased sensitivity to proteolysis and rapid intracellular degradation in mammalian cells. In conclusion, dmLT is a safe and powerful detoxified enterotoxin with the potential to function as a mucosal adjuvant for coadministered antigens and to elicit anti-LT antibodies without undesirable side effects.

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American Society for Microbiology

Tập 18 Số 4

546-551

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