Chimeric Viruses presenting non-native surface proteins and uses thereof
Patents Granted (Contd.) FORM 25 (12) PATENT (19) AP (11) Patent No : AP 2911 (73) Applicant(s) (21) Application No : AP/P/2008/004504 THE MOUNT SINAI SCHOOL OF MEDICINE OF NEW YORK UNIVERSITY (22) Filing Date: 01.12.2006 One Gustave L. Levy Place, New York, NY 10029-6574, United States of America (24) (45) Date of Grant & Publication : 05.05.2014 (30) Priority Data (33) Country (31) Number (32) Date (72) Inventors US 60/741,833 02.12.2005 PALESE Peter, United States of America US 60/802,864 22.05.2006 GARCIA-SASTRE Adolfo, United States of America (84) Designated State (74) Representative BW GM GH KE LS MW MZ NA SL FISHER CORMACK & BOTHA SD SZ TZ UG ZM ZW (51) International Classification: (54) Title Chimeric Viruses presenting non-native surface proteins and uses thereof (57) Abstract The present invention provides chimeric negative-stand RNA viruses that allov\ a subject, e.g., an avian, to be immunized against two infectious agents by using a single chimeric virus of the invention. In particular, the present invention provides chimeric influenza viruses engineered to express and incorporate into their virions a fusion protein comprising an ectodomain of a protein of an infectious agent and the transmembrane and cytoplasmic domain of an influenza virus protein. Such chimeric viruses induce an immune response against influenza virus and the infectious agent. The present invention also provides chimeric Newcastle Disease viruses {NDV) engineered to express and incorporate into their virions a fusion protein comprising the ectodomain of a protein of an infectious agent and the transmembrane and cytoplasmic domain of an NDV protein. Such chimeric viruses induce an immune response against NDV and the infectious agent. (56) Documents Cited : The present invention provides chimeric negative-stand RNA viruses that allow a subject, e.g., an avian, to be immunized against two infectious agents by using a single chimeric virus of the invention. In particular, the present invention provides chimeric Newcastle Disease viruses (NDV) engineered to express and incorporate into their virions a fusion protein comprising the ectodomain of a protein of an infectious agent and the transmembrane and cytoplasmic domain of an NDV protein. Such chimeric viruses induce an immune response against NDV and the infectious agent. A chimeric Newcastle Disease Virus (NDV), comprising a packaged genome comprising:
a. a nucleotide sequence encoding an F-fusion protein having the transmembrane and cytoplasmic domains of an F protein and at least one epitope of an ectodomain of a protective antigen of an infectious agent or an antigen associated with a disease that is anchored by the C-terminus, so that the F-fusion protein is expressed and incorporated into the chimeric NDV, and wherein the antigen is not a paramyxovirus antigen; or b. a nucleotide sequence encoding an HN-fusion protein having the transmembrane and cytoplasmic domains of an HN protein and at least one epitope of an ectodomain of a protective antigen of an infectious agent or an antigen associated with a disease that is anchored by the N-terminus, so that the HN-fusion protein is expressed and incorporated into the chimeric NDV, and wherein the antigen is not a paramyxovirus antigen. The chimeric NDV of claim 1 which comprises a nucleotide sequence encoding an F-fusion protein having the transmembrane and cytoplasmic domains of an F protein and at least one epitope of an ectodomain of a protective antigen of an infectious agent or an antigen associated with a disease that is anchored by the C-terminus, so that the F-fusion protein is expressed and incorporated into the chimeric NDV, and wherein the antigen is not a paramyxovirus antigen. The chimeric NDV of claim 1 which comprises a nucleotide sequence encoding an HN-fusion protein having the transmembrane and cytoplasmic domains of an HN protein and at least one epitope of an ectodomain of a protective antigen of an infectious agent or an antigen associated with a disease that is anchored by the N-terminus, so that the HN-fusion protein is expressed and incorporated into the chimeric NDV, and wherein the antigen is not a paramyxovirus antigen. The chimeric NDV of any one of claims 1 to 3 in which the genome comprises a nucleotide sequence encoding an HN protein, so that the HN protein is expressed and incorporated into the chimeric NDV. The chimeric NDV of any one of claims 1 to 4 in which the genome comprises a nucleotide sequence encoding an F protein, so that the F protein is expressed and incorporated into the chimeric NDV. The chimeric NDV of claim 1 or 2 in which the nucleotide sequence encoding the F-fusion protein replaces the nucleotide sequence encoding the NDV F protein and the F-fusion protein supplies the function of the F protein. The chimeric NDV of claim 1 or 3 in which the nucleotide sequence encoding the HN-fusion protein replaces the nucleotide sequence encoding the NDV HN protein and the HN-fusion protein supplies the function of the HN protein. The chimeric NDV of claim 1, 2, or 6, wherein the antigen is the ectodomain of a hemagglutinin antigen of an influenza virus. The chimeric NDV of claim 1, 3, or 7, wherein the antigen is the ectodomain of a neuraminidase antigen of an influenza virus. The chimeric NDV of claim 8 or 9, wherein the influenza virus is an avian influenza virus. The chimeric NDV of any one of the preceding claims, wherein the chimeric NDV is attenuated. The chimeric NDV of any one of claims 1 to 7, wherein the infectious agent is an infectious pathogen. An immunogenic composition comprising the chimeric NDV of any one of the preceding claims. The chimeric NDV of any one of claims 1 to 12 or the immunogenic composition of claim 13 for use in inducing an immune response to NDV and the antigen in an avian, or for use in inducing an immune response to the antigen in a human. A method for producing an immunogenic formulation, the method comprising:
a. propagating the chimeric NDV of any one of claims 1 to 12 in an embryonated egg or in a cell line that is susceptible to an NDV infection; and b. collecting the progeny virus, Use of the chimeric NDV of any one of claims 1 to 12 in the preparation of a medicament for use in inducing an immune response to NDV and the antigen in an avian. Use of the chimeric NDV of any one of claims 1 to 12 in the preparation of a medicament for use in inducing an immune response to the antigen in a human. A recombinant DNA molecule encoding the chimeric NDV of any one of claims 9 to 10.