Mutant CyaA Polypeptides and Polypeptide Derivatives Suitable for the Delivery of Immunogenic Molecules Into a Cell

A Polypeptide which is a mutant of an adenylate cyclase protein (mutant polypeptide) and whose amino acid sequence comprises or consists of one of the following sequences:

• a) the amino acid sequence of the adenylate cyclase (CyaA) of Bordetella pertussis, Bordetella parapertussis or Bordetella hinzii wherein the following mutations have been performed:
• (i) the substitution of the glutamic acid residue at position 570 by a glutamine residue (E570Q) or by a conservative amino acid residue, and
• (ii) the substitution of the lysine residue at position 860 by an arginine residue (K860R) or by a conservative amino acid residue, or;
• b) an amino acid sequence of a fragment of the adenylate cyclase of Bordetella pertussis, Bordetella parapertussis or Bordetella hinzii, which fragment has the capacity of the CyaA protein of Bordetella pertussis to bind to a target cell and the capacity to translocate its N-terminal adenylate cyclase enzyme domain or part thereof into said cell, wherein said fragment further contains the following mutated amino acid residues located at positions 570 and 860 in said adenylate cyclase:
• E570Q and K860R, or
• c) an amino acid sequence which differs from the amino acid sequence as defined in a) or b) by one or more amino acid residue substitutions and/or insertions and which has the capacity of the CyaA protein of Bordetella Pertussis to bind to a target cell and the capacity to translocate its N-terminal adenylate cyclase enzyme domain or part thereof into said cell, wherein said amino acid sequence further contains the following mutated amino acid residues located at positions 570 and 860 in said adenylate cyclase:
• E570Q and K860R, or
• d) the amino acid sequence of the adenylate cyclase (CyaA) of Bordetella bronchiseptica wherein the following mutations have been performed:
• (i) the substitution of the glutamic acid residue at position 569 by a glutamine residue (E569Q) or by a conservative amino acid residue, and
• (ii) the substitution of the lysine residue at position 859 by an arginine residue (K859R) or by a conservative amino acid residue, or;
• e) an amino acid sequence of a fragment of the adenylate cyclase of Bordetella bronchiseptica, which fragment has the capacity of the CyaA protein of Bordetella pertussis to bind to a target cell and the capacity to translocate its N-terminal adenylate cyclase enzyme domain or part thereof into said cell, wherein said fragment further contains the following mutated amino acid residues located at positions 569 and 859 in said adenylate cyclase:
• E569Q and K859R, or
• f) an amino acid sequence which differs from the amino acid sequence as defined in d) or e) by one or more amino acid residue substitutions and/or insertions and which has the capacity of the CyaA protein of Bordetella Pertussis to bind to a target cell and the capacity to translocate its N-terminal adenylate cyclase enzyme domain or part thereof into said cell, wherein said amino acid sequence further contains the following mutated amino acid residues located at positions 569 and 859 in said adenylate cyclase:
• E569Q and K859R.