Abstract
Heat-labile enterotoxin (LT) from Escherichia coli and Cholera toxin (CT) from Vibro cholerae are known to be potent mucosal immunogens. These toxins have 80% sequence homology and a similar tertiary structure (1,4), and both elicit potent serum IgG and mucosal IgA responses (5,6). Moreover, both also serve as excellent adjuvants for coadministered antigens. However, they are toxic in their native state and both produce accumulation of intestinal fluid and watery diarrhea (7). LT is the cause of traveler’s diarrhea, whereas CT causes cholera. In order to make use of the adjuvanticity of these molecules but reduce their toxicity, several mutants have been generated by site-directed mutagenesis. Of these, there are two mutants of the enzymatic A subunit, LTK63 and LTR72, that maintain a high level of immunogenicity and have significant potential as adjuvants. This chapter will focus on the use of LTK63 and LTR72 as intramuscular and intranasal adjuvants for the induction of cytotoxic T lymphocytes (CTL) activity against coadministered protein immunogens.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Magagnoli, C., Manetti, R., Fontana, M. R., Giannelli, V., Giuliani, M. M., Rappuoli, R., et al. (1996) Mutations in the A subunit affect yield, stability, and protease sensitivity of nontoxic derivatives of heat labile enterotoxin. Infection and Immunity 64, 5434–5438.
Dallas, W. S. and Falkow, S. (1980) Amino acid homology between cholera toxin and Escherichia coli heat labile toxin. Nature 288, 499–501.
Spicer, E. K., Kavanaugh, W. M., Dallas, W. S., Falkow, S., Konisberg, W. H., and Shafer, D. (1981) Sequence homologies between A subunits of E. Coli and V. cholerae enterotoxin. Proc. Natl. Acad. Sci. USA 78, 50–54.
Yamamoto, T., Tamura, T., and Yokota, T. (1984) Primary structure of heat-labile enterotoxin produced by Escherichia coli pathogenic for humans. J. Biol. Chem. 259, 5037–5044.
Clements, J. D. and Dickinson, B. L. (1996) Use of Escherichia coli heat-labile enterotoxin as an oral adjuvant, in Mucosal Vaccines (Kiyono, H., Ogra, P. L., and McGhee, J. R., eds.), Academic, London and New York, pp. 73–87.
Douce, G., Turcotte, C., Cropley, I., Roberts, M., Pizza, M., Domenighini, M., et al. (1995) Mutants of Escherichia coli heat-labile toxin lacking ADP-ribosyltransferase activity act as nontoxic, mucosal adjuvants. Proc. Natl. Acad. Sci. USA 92, 1644–1648.
Pizza, M., Fontana, M. R., Giuliani, M. M., Domenighini, M., Magagnoli, C., Giannelli, V., et al. (1994) A genetically detoxified derivative of heat-labile Escherichia coli enterotoxin induces neutralizing antibodies against the A sub-unit. J. Exp. Med. 180, 2147–2153.
Holmgren, J., Lonnroth, I., and Svennerholm, L. (1973) Tissue receptor for cholera exotoxin: postulated structure from studies with GM1-ganglioside and related glycolipids. Infect. Immun. 8, 208–214.
Sugii, S. T. (1989) Binding specificities of heat-labile enterotoxins isolated from porcine and human enterotoxigenic Escherichia coli heat-labile toxin and cholera toxin. Can. J. Microbiol. 35, 670–673.
Pizza, M., Fontana, M. R., Scarlato, V., and Rappuoli, R. (1996) Genetic detoxification of bacterial toxins in, Methods in Molecular Medicine: Vaccine Protocols (Robinson, A., Farrar, G., and Wiblin, C., eds.), Humana, Totowa, NJ, pp. 91–109.
Lycke, N. and Holmgren, J. (1992) The adjuvant effect of Vibro cholerae and Escherichia coli heat-labile enterotoxins is linked to ADP-ribosyltransferase activity. Eur. J. Immunol. 22, 2277.
Di Tommaso, A., Saletti, G., Pizza, M., Rappuoli, R., Dougan, G., Abrignani, S., et al. (1996) Induction of antigen specific antibodies in vaginal secretions by using a nontoxic mutant heat-labile enterotoxin as a mucosal adjuvant. Infect. Immun. 64, 974–979.
Partidos, C. D., Pizza, M., Rappuoli, R., and Steward, M. W. (1996) The adjuvant effect of a non-toxic mutant heat-labile enterotoxin of Escherichia coli for the induction of measles virus-specific CTL responses after intranasal co-immunization with a synthetic peptide. Immunology 89, 483–487.
Clements, J. D. and Finkelstein, R. A. (1979) Isolation and characterization of homogeneous heat-labile enterotoxin with high specific activity from Escherichia coli cultures. Infect. Immun. 24, 760–769.
Clements, J. D., Yancy, R. J., and Finkelstein, R. A. (1980) Properties of homogeneous heat-labile enterotoxin from Escherichia coli. Infect. Immun. 29, 91–97.
Pizza M., Domenighini M., Hoi, W., Giannelli, V., Fontana, M. R., Giuliani, M. M., et al. (1994) Probing the structure-activity relationship of Escherichia coli LT-A by site-directed mutagenesis. Mol. Microb. 14, 51–60.
Pizza, M., Fontana, M. R., Giuliani, M. M., Domenighini, M., Magagnoli, C., Giannelli, V., et al. (1994) A genetically detoxified derivative of heat-labile Escherichia coli enterotoxin induces neutralizing antibodies against the A sub-unit. J. Exp. Med. 180, 2147–2153.
Giuliani, M. M., Del Giudice, G., Giannelli, V., Dougan, G., Douce, D., Rappuoli, R., et al. (1998) Mucosal adjuvanticity and immunogenicity of LTR72, a novel mutant of Escherichia coli heat-labile enterotoxin with partial knockout of ADP-ribosyltransferase activity. J. Exp. Med. 187, 1–10.
Doe, B. and Walker, C. M. (1996) HIV-1 p24 gag-specific cytotoxic T-lymphocyte responses in mice. Correspondence, AIDS 10, 793–806.
Elvin, J., Cerundolo, V., Elliott, T., and Townsend, A. (1991) A quantitative assay of peptide-dependant class I assembly. Eur. J. Immunol. 21, 2025–2031.
Paliard, X., Doe, B., and Walker, C. M. (1998) The T cell repertoire primed by antiviral vaccination is influenced by self-tolerance. Cellular Immunol. 188, 73–79.
Doe, B., Steimer, K. S., and Walker, C. M. (1994) Induction of HIV-1 envelope (gp120)-specific cytotoxic T lymphocyte responses in mice by recombinant CHO-derived gp120 is enhanced by enzymatic removal of N-linked glycans. Euro. J. Immunol. 24, 2369–2376.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc.
About this protocol
Cite this protocol
Neidleman, J.A., Ott, G., O’Hagan, D. (2000). Mutant Heat-Labile Entertoxins As Adjuvants for CTL Induction. In: O’Hagan, D.T. (eds) Vaccine Adjuvants. Methods in Molecular Medicine™, vol 42. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-083-7:327
Download citation
DOI: https://doi.org/10.1385/1-59259-083-7:327
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-735-9
Online ISBN: 978-1-59259-083-4
eBook Packages: Springer Protocols