title: A mutant pertussis toxin molecule that lacks ADP-ribosyltransferase activity, PT-9K/129G, is an effective mucosal adjuvant for intranasally delivered proteins.
abstract: We examined the capacity of a genetically detoxified derivative of pertussis toxin (PTX), PT-9K/129G, to act as a mucosal adjuvant for an intranasally (i.n.) administered tetanus vaccine. Groups of mice were immunized i.n. with the nontoxic C-terminal 50-kDa portion of tetanus toxin (fragment C [Frg C]) either alone or mixed with PT-9K/129G, PTX, or cholera toxin (CT) or were immunized subcutaneously (s.c.) with an equivalent amount of Frg C adsorbed to alhydrogel. In response to a single immunization, mice receiving Frg C plus PT-9K/129G or CT i.n. and parenterally immunized mice developed high-titer (> 20,000) anti-Frg C antibodies, whereas mice immunized i.n. with Frg C plus PTX or with Frg C alone seroconverted only after being boosted. The serum anti-Frg C response was dominated by immunoglobulin G1 (IgG1) in mice immunized with Frg C plus PT-9K/129G, with Frg C plus PTX, or s.c. In contrast, IgG1, IgG2a, and IgG2b contributed almost equally to the Frg C response when CT was the adjuvant. Anti-Frg C IgE was detected only in the sera of mice immunized i.n. with Frg C plus PTX and immunized s.c. with Frg C plus alhydrogel. High levels of IgA antibodies were present in nasal lavage fluid from mice immunized i.n. with Frg C plus PT-9K/129G, PTX, or CT but not in that from mice given Frg C alone i.n. or parenterally. The mucosal adjuvanticity of PT-9K/129G was manifested in inbred as well as outbred mice. A single i.n. dose of Frg C plus either PT-9K/129G or PTX (with high specific activity) was sufficient to protect all immunized mice from tetanus toxin challenge, in contrast to the case for mice that received Frg C alone i.n. We conclude that the pertussis toxin analog PT-9K/129G, which is devoid of ADP-ribosyltransferase activity, is a potent mucosal adjuvant for vaccines delivered via the respiratory tract.
authors.affiliation: Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, United Kingdom.
authors.forename: M
authors.initials: M
authors.lastname: Roberts
authors.forename: A
authors.initials: A
authors.lastname: Bacon
authors.forename: R
authors.initials: R
authors.lastname: Rappuoli
authors.forename: M
authors.initials: M
authors.lastname: Pizza
authors.forename: I
authors.initials: I
authors.lastname: Cropley
authors.forename: G
authors.initials: G
authors.lastname: Douce
authors.forename: G
authors.initials: G
authors.lastname: Dougan
authors.forename: M
authors.initials: M
authors.lastname: Marinaro
authors.forename: J
authors.initials: J
authors.lastname: McGhee
authors.forename: S
authors.initials: S
authors.lastname: Chatfield
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000281:Administration, Intranasal; D000818:Animals; D002772:Cholera Toxin / Q000276:immunology; D005260:Female; D007114:Immunization; D007071:Immunoglobulin A, Secretory / Q000032:analysis; D007073:Immunoglobulin E / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000276:immunology*; D037342:Pertussis Toxin; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D013744:Tetanus Toxin / Q000276:immunology*; D010566:Virulence Factors, Bordetella / Q000276:immunology / Q000494:pharmacology*
chemical_list: D000276:Adjuvants, Immunologic; D007071:Immunoglobulin A, Secretory; D007074:Immunoglobulin G; D010446:Peptide Fragments; D013744:Tetanus Toxin; D010566:Virulence Factors, Bordetella; C040635:tetanus toxin fragment C; D007073:Immunoglobulin E; D002772:Cholera Toxin; D011065:Poly(ADP-ribose) Polymerases; D037342:Pertussis Toxin