The C-terminal domain is essential for protective activity of the Bordetella pertussis adenylate cyclase-hemolysin.
The adenylate cyclase-hemolysin of Bordetella pertussis consists of a cell-invasive N-terminal adenylate cyclase domain linked to a C-terminal RTX hemolysin containing extensive glycine-rich repeats. The toxin is an essential virulence factor required in the initial stages of infection. Adenylate cyclase-hemolysin was also shown to be a potent vaccinating antigen inducing protection against B. pertussis colonization of the mouse respiratory tract. This protective activity depends on a posttranslational fatty-acylation modification. We used a set of deletion derivatives of the recombinant adenylate cyclase-hemolysin to localize the protective epitopes on the 1,706-residue toxin. We show that specific anti-adenylate cyclase-hemolysin antibodies present in the sera of B. pertussis-infected mice and humans are directed predominantly against the modification-and-repeat portion of the toxin, contained in the last 800 residues of the adenylate cyclase-hemolysin. These antibodies appear to recognize conformational epitopes present only in a structure formed by the intact C-terminal half of the toxin. There was no correlation between the capacity of the truncated adenylate cyclase-hemolysin derivatives to induce both toxin-neutralizing antibodies upon immunization of mice and protective immunity. However, only the truncated proteins which were recognized by the sera of infected mice and humans and which had their last 800 residues intact had the capacity to induce protection of mice against colonization by B. pertussis. This indicates that the structure of the modification-and-repeat region of adenylate cyclase-hemolysin is critical for its protective activity.
Betsou
F
F
Unité de Bactériologie Moléculaire et Médicale, Institut Pasteur, Paris, France.
Sebo
P
P
Guiso
N
N
D000262:Adenylyl Cyclases / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis; D001886:Bordetella pertussis / Q000276:immunology*; D002675:Child, Preschool; D005260:Female; D006460:Hemolysin Proteins / Q000276:immunology*; D006801:Humans; D007114:Immunization; D007223:Infant; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D013329:Structure-Activity Relationship
D000907:Antibodies, Bacterial; D006460:Hemolysin Proteins; D011993:Recombinant Fusion Proteins; D000262:Adenylyl Cyclases