# US 7,611,714 B2 · Methods and compositions for immunizing against Pseudomonas infection > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,611,714 B2 - Document ID: US7611714B2 - Status: Granted - Kind: B2 - Application number: 11/244,348 - Filed: 2005-10-04 - Published: 2006-05-18 - Granted: 2009-11-03 - Earliest claimed: 2004-10-04 - Assignees: Trinity Biosystems, Inc. - Inventors: Randall J. Mrsny - Web page: https://patentgrep.com/patent/US7611714B2 ## CPC - C07K 14/21: from Pseudomonadaceae (F) - A61K 39/104: {Pseudomonadales, e.g.} Pseudomonas - C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation - C07K 2319/55: containing a fusion with a toxin, e.g. diphteria toxin - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) ## Abstract Methods and compositions for inducing an immune response against Pseudomonas aeruginosa are provided herein. In one aspect, the invention provides a chimeric immunogen, comprising a receptor binding domain, a translocation domain, and a Pseudomonas pilin peptide comprising an amino acid sequence that is TAADGLWKCTSDQDEQFIPKGCSK (SEQ ID NO.:1), wherein the chimeric immunogen, when administered to a subject, induces an immune response in said subject that is effective to reduce adherence of a microorganism that expresses said Pseudomonas pilin peptide to epithelial cells of said subject. In other aspects, the invention provides nucleic acids encoding chimeric immunogens of the invention, kits comprising chimeric immunogens of the invention, cells expressing chimeric immunogens of the invention, and methods of using chimeric immunogens of the invention. ## Claims ### Claim 1 A chimeric immunogen, comprising a)—a receptor binding domain, b)—a translocation domain, and c)—a Pseudomonas pilin peptide consisting of the amino acid sequence that is TAADGLWKCTSDQDEQFIPKGCSK (SEQ ID NO.: 1), wherein said chimeric immunogen, when administered to a subject, induces an immune response in said subject that is effective to reduce adherence of a Pseudomonas bacterium that expresses said Pseudomonas pilin peptide to epithelial cells of said subject. ### Claim 2 (depends on 1) The chimeric immunogen of claim 1, wherein said chimeric immunogen, when administered to said subject, generates an immune response in said subject that reduces the cytotoxicity of Pseudomonas exotoxin A to the subject. ### Claim 3 (depends on 1) The chimeric immunogen of claim 1, wherein said chimeric immunogen further comprises an endoplasmic reticulum retention domain. ### Claim 4 (depends on 3) The chimeric immunogen of claim 3, wherein said Pseudomonas pilin peptide is located between said translocation domain and said endoplasmic reticulum retention domain. ### Claim 5 (depends on 3) The chimeric immunogen of claim 3, wherein said endoplasmic reticulum retention domain is an enzymatically inactive domain III of Pseudomonas exotoxin A. ### Claim 6 (depends on 5) The chimeric immunogen of claim 5, wherein said enzymatically inactive domain III of Pseudomonas exotoxin A is inactivated by deleting a glutamate at position 553. ### Claim 7 (depends on 3) The chimeric immunogen of claim 3, wherein said endoplasmic reticulum retention domain comprises an amino acid sequence that is selected from the group of RDEL (SEQ ID NO.:2) or KDEL (SEQ ID NO.:3). ### Claim 8 (depends on 1) The chimeric immunogen of claim 1, wherein said translocation domain is selected from the group consisting translocation domains from Pseudomonas exotoxin A, diptheria toxin, pertussis toxin, cholera toxin, heat-labile E. coli enterotoxin, shiga toxin, and shiga-like toxin. ### Claim 9 (depends on 5) The chimeric immunogen of claim 5, wherein said translocation domain is domain II of Pseudomonas exotoxin A. ### Claim 10 (depends on 1) The chimeric immunogen of claim 1, wherein said translocation domain comprises amino acids 280 to 364 of domain II of Pseudomonas exotoxin A. ### Claim 11 (depends on 1) The chimeric immunogen of claim 1, wherein said chimeric immunogen comprises more than one of said Pseudomonas pilin peptides. ### Claim 12 (depends on 1) The chimeric immunogen of claim 1, wherein said receptor binding domain is selected from the group consisting of domain Ia of Pseudomonas exotoxin A; a receptor binding domains from cholera toxin, diptheria toxin, shiga toxin, or shiga-like toxin; a monoclonal antibody, a polyclonal antibody, or a single-chain antibody; TGFα, TGFβ, EGF, PDGF, IGF, or FGF; IL-1, IL-2, IL-3, or IL-6; and MIP-1a, MIP-1b, MCAF, or IL-8. ### Claim 13 (depends on 12) The chimeric immunogen of claim 12, wherein said receptor binding domain is domain Ia of Pseudomonas exotoxin A. ### Claim 14 (depends on 13) The chimeric immunogen of claim 13, wherein said domain Ia of Pseudomonas exotoxin A has an amino acid sequence that is SEQ ID NO.:4. ### Claim 15 (depends on 1) The chimeric immunogen of claim 1, wherein said receptor binding domain binds to α2-macroglobulin receptor, epidermal growth factor receptor, transferrin receptor, interleukin-2 receptor, interleukin-6 receptor, interleukin-8 receptor, Fc receptor, poly-IgG receptor, asialoglycopolypeptide receptor, CD3, CD4, CD8, chemokine receptor, CD25, CD11B, CD11C, CD80, CD86, TNFα receptor, TOLL receptor, M-CSF receptor, GM-CSF receptor, scavenger receptor, or VEGF receptor. ### Claim 16 (depends on 15) The chimeric immunogen of claim 15, wherein said receptor binding domain binds to α2-macroglobulin receptor. ### Claim 17 (depends on 1) The chimeric immunogen of claim 1, wherein said chimeric immunogen has an amino acid sequence that is SEQ ID NO.:5. ### Claim 18 A composition comprising a chimeric immunogen, wherein said chimeric immunogen comprises: a)—a receptor binding domain, b)—a translocation domain, and c)—a Pseudomonas pilin peptide that has an amino acid sequence that is TAADGLWKCTSDQDEQFIPKGCSK (SEQ ID NO.: 1) wherein said chimeric immunogen, when administered to a subject, induces an immune response in said subject that is effective to reduce adherence of a Pseudomonas bacterium that expresses said Pseudomonas pilin peptide to epithelial cells of said subject. ### Claim 19 (depends on 18) The composition of claim 18, wherein said composition further comprises a pharmaceutically acceptable diluent, excipient, vehicle, or carrier. ### Claim 20 (depends on 18) A kit comprising the composition of claim 18, wherein said composition is in a single-unit dosage form. ### Claim 21 A method for inducing an immune response in a subject, comprising administering to said subject an effective amount of a chimeric immunogen comprising a receptor binding domain, a translocation domain, and a Pseudomonas pilin peptide that consists of the amino acid sequence that is TAADGLWKCTSDQDEQFIPKGCSK (SEQ ID NO.: 1), wherein said chimeric immunogen induces an immune response in said subject that is effective to reduce adherence of a microorganism expressing the Pseudomonas pilin peptide to epithelial cells of said subject when said chimeric immunogen is administered to said subject. ### Claim 22 A method for generating in a subject antibodies specific for a Pseudomonas pilin peptide consisting of the amino acid sequence that is TAADGLWKCTSDQDEQFIPKGCSK (SEQ ID NO.:1), comprising administering to said subject an effective amount of a chimeric immunogen comprising a receptor binding domain, a translocation domain, and a Pseudomonas pilin peptide that consists of the amino acid sequence that is TAADGLWKCTSDQDEQFIPKGCSK (SEQ ID NO.: 1), thereby generating antibodies specific for said Pseudomonas pilin peptide.