Mutants of Escherichia coli heat-labile toxin lacking ADP-ribosyltransferase activity act as nontoxic, mucosal adjuvants

G. Douce; C. Turcotte; I. Cropley; M. Roberts; M. Pizza; R. Rappuoli; G. Dougan

doi:10.1073/pnas.92.5.1644

Mutants of Escherichia coli heat-labile toxin lacking ADP-ribosyltransferase activity act as nontoxic, mucosal adjuvants

G. Douce
, C. Turcotte
, I. Cropley
, M. Roberts
, M. Pizza
, R. Rappuoli
, G. Dougan

Research output: Contribution to journal › Article › Research › peer-review

250 Citations (Scopus)

Abstract

A nontoxic mutant (LTK7) of the Escherichia coli heat-labile enterotoxin (LT) lacking ADP-ribosylating activity but retaining holotoxin formation was constructed. By using site-directed mutagenesis, the arginine at position 7 of the A subunit was replaced with lysine. This molecule, which was nontoxic in several assays, was able to bind to eukaryotic cells and acted as a mucosal adjuvant for co-administered proteins; BALB/c mice immunized intranasally with LTK7 and ovalbumin developed high levels of serum and local antibodies to ovalbumin and toxin. In addition, mice immunized intranasally with fragment C of tetanus toxin and LTK7 were protected against lethal challenge with tetanus toxin. Thus nontoxic mutants of heat-labile toxin can act as effective intranasal mucosal adjuvants.

Original language	English
Pages (from-to)	1644-1648
Number of pages	5
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	92
Issue number	5
DOIs	https://doi.org/10.1073/pnas.92.5.1644
Publication status	Published - 28 Feb 1995
Externally published	Yes

Keywords

Heat-labile enterotoxin
Nontoxic mutant
Tetanus

Access to Document

10.1073/pnas.92.5.1644

Cite this

@article{0296fa7bf2e64131a527fed11abef217,

title = "Mutants of Escherichia coli heat-labile toxin lacking ADP-ribosyltransferase activity act as nontoxic, mucosal adjuvants",

abstract = "A nontoxic mutant (LTK7) of the Escherichia coli heat-labile enterotoxin (LT) lacking ADP-ribosylating activity but retaining holotoxin formation was constructed. By using site-directed mutagenesis, the arginine at position 7 of the A subunit was replaced with lysine. This molecule, which was nontoxic in several assays, was able to bind to eukaryotic cells and acted as a mucosal adjuvant for co-administered proteins; BALB/c mice immunized intranasally with LTK7 and ovalbumin developed high levels of serum and local antibodies to ovalbumin and toxin. In addition, mice immunized intranasally with fragment C of tetanus toxin and LTK7 were protected against lethal challenge with tetanus toxin. Thus nontoxic mutants of heat-labile toxin can act as effective intranasal mucosal adjuvants.",

keywords = "Heat-labile enterotoxin, Nontoxic mutant, Tetanus",

author = "G. Douce and C. Turcotte and I. Cropley and M. Roberts and M. Pizza and R. Rappuoli and G. Dougan",

year = "1995",

month = feb,

day = "28",

doi = "10.1073/pnas.92.5.1644",

language = "English",

volume = "92",

pages = "1644--1648",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "National Academy of Sciences",

number = "5",

}

Mutants of Escherichia coli heat-labile toxin lacking ADP-ribosyltransferase activity act as nontoxic, mucosal adjuvants. / Douce, G.; Turcotte, C.; Cropley, I. et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, No. 5, 28.02.1995, p. 1644-1648.

Research output: Contribution to journal › Article › Research › peer-review

TY - JOUR

T1 - Mutants of Escherichia coli heat-labile toxin lacking ADP-ribosyltransferase activity act as nontoxic, mucosal adjuvants

AU - Douce, G.

AU - Turcotte, C.

AU - Cropley, I.

AU - Roberts, M.

AU - Pizza, M.

AU - Rappuoli, R.

AU - Dougan, G.

PY - 1995/2/28

Y1 - 1995/2/28

N2 - A nontoxic mutant (LTK7) of the Escherichia coli heat-labile enterotoxin (LT) lacking ADP-ribosylating activity but retaining holotoxin formation was constructed. By using site-directed mutagenesis, the arginine at position 7 of the A subunit was replaced with lysine. This molecule, which was nontoxic in several assays, was able to bind to eukaryotic cells and acted as a mucosal adjuvant for co-administered proteins; BALB/c mice immunized intranasally with LTK7 and ovalbumin developed high levels of serum and local antibodies to ovalbumin and toxin. In addition, mice immunized intranasally with fragment C of tetanus toxin and LTK7 were protected against lethal challenge with tetanus toxin. Thus nontoxic mutants of heat-labile toxin can act as effective intranasal mucosal adjuvants.

AB - A nontoxic mutant (LTK7) of the Escherichia coli heat-labile enterotoxin (LT) lacking ADP-ribosylating activity but retaining holotoxin formation was constructed. By using site-directed mutagenesis, the arginine at position 7 of the A subunit was replaced with lysine. This molecule, which was nontoxic in several assays, was able to bind to eukaryotic cells and acted as a mucosal adjuvant for co-administered proteins; BALB/c mice immunized intranasally with LTK7 and ovalbumin developed high levels of serum and local antibodies to ovalbumin and toxin. In addition, mice immunized intranasally with fragment C of tetanus toxin and LTK7 were protected against lethal challenge with tetanus toxin. Thus nontoxic mutants of heat-labile toxin can act as effective intranasal mucosal adjuvants.

KW - Heat-labile enterotoxin

KW - Nontoxic mutant

KW - Tetanus

UR - https://www.scopus.com/pages/publications/0028918767

U2 - 10.1073/pnas.92.5.1644

DO - 10.1073/pnas.92.5.1644

M3 - Article

C2 - 7878032

AN - SCOPUS:0028918767

SN - 0027-8424

VL - 92

SP - 1644

EP - 1648

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 5

ER -

Mutants of Escherichia coli heat-labile toxin lacking ADP-ribosyltransferase activity act as nontoxic, mucosal adjuvants

Abstract

Keywords

Access to Document

Other files and links

Cite this