Alphavirus Vectors for Respiratory Pathogen Vaccines

Abstract

Described herein are compositions and methods for stimulating an immune response to one or more proteins derived from one or more respiratory pathogens. In particular, the invention relates to alphavirus replicons, alphavirus vector constructs, alphavirus replicon particles expressing one or more antigens derived from one or more respiratory pathogens as well as to method of making and using these immunogenic compositions.

1 . An immunogenic composition comprising
 (a) a first alphavirus replicon vector, vector construct or replicon particle comprising:   (i) a first heterologous nucleic acid encoding at least one immunogenic protein derived from a respiratory pathogen; and   (ii) a second heterologous nucleic acid encoding at least one immunogenic protein derived from a respiratory pathogen;   wherein if the first and second heterologous nucleic acids both encode an immunogenic protein derived from the same parainfluenza virus (PIV), the same respiratory syncytial virus (RSV) or the same SARS virus, the composition further comprises a second alphavirus replicon vector, vector construct or replicon particle comprising the second heterologous nucleic acid; and   (b) a pharmaceutically acceptable carrier, diluent, or excipient.   
     
     
         2 . The immunogenic composition of  claim 1 , wherein the first alphavirus replicon vector, vector construct or replicon particle encodes the first and second heterologous nucleic acids. 
     
     
         3 . The immunogenic composition of  claim 1 , further comprising a second alphavirus replicon vector, vector construct or replicon particle, wherein the first alphavirus replicon vector, vector construct or replicon particle comprises the first heterologous nucleic acid sequences and the second alphavirus replicon vector, vector construct or replicon particle comprises the second heterologous nucleic acid. 
     
     
         4 . The immunogenic composition of  claim 1 , wherein the respiratory pathogen is a virus selected from the group consisting of an influenza virus, a parainfluenza virus, a respiratory syncytial virus, a human metapneumovirus and a SARS virus. 
     
     
         5 . The immunogenic composition of  claim 1 , wherein the respiratory pathogen is a bacteria selected from the group consisting of  Mycobacterium tuberculosis; Corynebacterium diphtheriae; Bordatella pertussis; Streptococcus pneumoniae ; nontypeable  Haemophilus influenzae; Moraxella catarrhalis; Pseudomonas aeruginosa; Bacillus anthracis  (anthrax); and  Legionella pneumophila  (Legionnaires' Disease). 
     
     
         6 . The immunogenic composition according to  claim 4 , wherein the first heterologous nucleic acid encodes an influenza virus neuraminidase protein derived from an influenza virus subtype selected from the group consisting of N1, N2, N3, N4, N5, N6, N7, N8, and N9. 
     
     
         7 . The immunogenic composition according to  claim 4 , wherein the first heterologous nucleic acid encodes an influenza virus neuraminidase protein and the alphavirus replicon vector, vector construct or replicon particle further comprises a second heterologous nucleic acid encoding an influenza virus hemagglutinin protein. 
     
     
         8 . The immunogenic composition according to  claim 7 , wherein the second heterologous nucleic acid encodes a hemagglutinin protein derived from an influenza virus subtype selected from the group consisting of H1, H2H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, and H15. 
     
     
         9 . The immunogenic composition according to  claim 1 , wherein the first heterologous nucleic acid is operably linked to a first alphavirus subgenomic junction region promoter, and the second heterologous nucleic acid is operably linked to a second alphavirus subgenomic junction region promoter. 
     
     
         10 . The immunogenic composition according to  claim 1 , wherein the first or second heterologous nucleic acid further comprises a nucleic acid corresponding to an internal ribosome entry site (IRES). 
     
     
         11 . The immunogenic composition according to  claim 1 , wherein the first heterologous nucleic acid encodes an influenza virus protein and the second heterologous nucleic acid encodes a SARS coronavirus protein. 
     
     
         12 . The immunogenic composition according to  claim 1 , wherein the first heterologous nucleic acid encodes a parainfluenza virus protein and the second heterologous nucleic acid encodes a respiratory syncytial virus protein. 
     
     
         13 . The immunogenic composition according to  claim 1 , wherein the first heterologous nucleic acid encodes a respiratory syncytial virus protein and the second heterologous nucleic acid encodes a human metapneumovirus protein. 
     
     
         14 . An immunogenic composition according to  claim 1  further comprising an adjuvant. 
     
     
         15 . The immunogenic composition according to  claim 1 , wherein the first or second heterologous nucleic acid encoding an influenza virus protein is selected from the group consisting of a sequence encoding a hemagglutinin protein, a sequence encoding a neuraminidase protein, a sequence encoding a nucleocapsid protein, and a sequence encoding a matrix protein. 
     
     
         16 . The immunogenic composition according to  claim 1 , wherein the first or second heterologous nucleic acid encoding a SARS coronavirus protein is selected from the group consisting of a nucleic acid encoding a spike protein, a nucleic acid encoding an envelope protein, a nucleic acid encoding a nucleocapsid protein, and a nucleic acid encoding a matrix protein. 
     
     
         17 . The immunogenic composition according to  claim 1 , wherein the first or second heterologous nucleic acid encoding a human metapneumovirus protein is selected from the group consisting of a nucleic acid encoding a glycoprotein G, a nucleic acid encoding a fusion protein, a nucleic acid encoding a nucleocapsid protein, and a nucleic acid encoding a matrix protein. 
     
     
         18 . The immunogenic composition according to  claim 1 , wherein the first or second heterologous nucleic acid encoding a parainfluenza virus protein is selected from the group consisting of a nucleic acid encoding a hemagglutinin-neuraminidase protein, a nucleic acid encoding a fusion protein, a nucleic acid encoding a nucleocapsid protein, and a nucleic acid encoding a matrix protein. 
     
     
         19 . The immunogenic composition according to  claim 1 , wherein the first or second heterologous nucleic acid encoding a respiratory syncytial virus protein is selected from the group consisting of a nucleic acid encoding a glycoprotein G, a nucleic acid encoding a fusion protein, a nucleic acid encoding a nucleocapsid protein, and a nucleic acid encoding a matrix protein. 
     
     
         20 . The immunogenic composition according to  claim 1 , wherein the first alphavirus replicon vector, vector construct and replicon particle is derived from one or more alphaviruses selected from the group consisting of a Sindbis virus, a Semliki Forest virus, a Venezuelan equine encephalitis virus, and a Ross River virus. 
     
     
         21 . The immunogenic composition according to  claim 3 , wherein the second alphavirus replicon vector, vector construct and replicon particle is derived from one or more alphaviruses selected from the group consisting of a Sindbis virus, a Semliki Forest virus, a Venezuelan equine encephalitis virus, and a Ross River virus. 
     
     
         22 . The immunogenic composition according to  claim 1 , wherein the immunogenic composition is lyophilized. 
     
     
         23 . A method of stimulating an immune response in a mammal, the method comprising administering an immunogenic composition according to  claim 1  the mammal, thereby generating an immune response. 
     
     
         24 . The method according to  claim 23 , further comprising the step of administering a second immunogenic composition to the mammal, wherein the second immunogenic composition comprises:
 a) a protein, polypeptide or portion thereof, derived from substantially the same source as the heterologous nucleic acid(s); and   b) a pharmaceutically acceptable carrier, diluent, or excipient.   
     
     
         25 . The method according to  claim 24 , wherein the step of administering a second immunogenic composition, further comprises administering an adjuvant. 
     
     
         26 . The method according to  claim 24 , wherein the second immunogenic composition further comprises an adjuvant. 
     
     
         27 . The method according to  claim 23 , wherein the first or second immunogenic compositions are administered by a route selected from the group consisting of intramuscular, intranasal, subcutaneous, intradermal, intratracheal, and oral. 
     
     
         28 . The method of stimulating an immune response according to  claim 23 , further comprising a second step of administering a second immunogenic composition, wherein the second immunogenic composition comprises:
 a) a non-alphavirus derived viral vector encoding a protein, polypeptide or portion thereof, derived from substantially the same source as the heterologous nucleic acid(s); and   b) a pharmaceutically acceptable carrier, diluent, or excipient.   
     
     
         29 . The method of stimulating an immune response according to  claim 23 , further comprising a second step of administering a second immunogenic composition, wherein the second immunogenic composition comprises:
 a) an attenuated virus encoding a protein, polypeptide or portion thereof, derived from substantially the same source as the heterologous nucleic acid(s); and   b) a pharmaceutically acceptable carrier, diluent, or excipient.   
     
     
         30 . (canceled) 
     
     
         31 . An immunogenic composition, comprising:
 a) a preparation of substantially purified, inactivated parainfluenza virus;   b) a pharmaceutically acceptable carrier, diluent, or excipient; and   c) an adjuvant selected from MF59 or LTK63.   
     
     
         32 . The immunogenic composition according to  claim 31 , wherein the adjuvant is MF59. 
     
     
         33 . A method of stimulating an immune response in a mammal, the method comprising administering an immunogenic composition according to  claim 31  to the mammal, thereby generating an immune response. 
     
     
         34 . The method according to  claim 33 , wherein the immunogenic composition is administered by a route selected from the group consisting of intramuscular, intranasal, subcutaneous, intradermal, intratracheal, and oral. 
     
     
         35 . An immunogenic composition comprising
 (a) a first alphavirus replicon vector, vector construct or replicon particle comprising a first heterologous nucleic acid encoding a first immunogenic neuraminidase protein derived from an influenza virus; and   (b) a pharmaceutically acceptable carrier, diluent, or excipient.   
     
     
         36 . The immunogenic composition of  claim 35 , further comprising
 a second heterologous nucleic acid encoding a second immunogenic protein derived from an influenza virus selected from the group consisting of: a hemagglutinin protein, nucleocapsid, matrix protein and a second neuraminidase protein.   
     
     
         37 . The immunogenic composition of  claim 36 , wherein the second immunogenic protein is a hemagglutinin protein derived from one or more pandemic or potentially pandemic influenza virus strains. 
     
     
         38 . The immunogenic composition of  claim 36 , wherein the second immunogenic protein is a hemagglutinin protein derived from one or more interpandemic influenza virus strains. 
     
     
         39 . The immunogenic composition of  claim 36 , wherein the second immunogenic protein is a hemagglutinin protein derived from a combination of one or more pandemic or potentially pandemic influenza virus strains and one or more interpandemic influenza virus strains 
     
     
         40 . The immunogenic composition of  claim 36 , wherein the first alphavirus replicon vector, vector construct or replicon particle comprises the second heterologous nucleic acid. 
     
     
         41 . The immunogenic composition of  claim 36 , further comprising a second alphavirus replicon vector, vector construct or replicon particle that comprises the second heterologous nucleic acid. 
     
     
         42 . The immunogenic composition of  claim 4 , wherein said respiratory pathogen is an influenza virus, said first heterologous nucleic acid encodes a hemagglutinin protein derived from a first influenza virus strain and said second heterologous nucleic acid encodes a hemagglutinin protein derived from a second influenza virus strain. 
     
     
         43 . The immunogenic composition of  claim 42  further comprising one or more additional heterologous nucleic acid encoding one or more additional immunogenic proteins derived from one or more additional influenza virus strains. 
     
     
         44 . The immunogenic composition of  claim 42  or  claim 43 , wherein the hemagglutinin proteins are derived from one or more pandemic or potentially pandemic influenza virus strains. 
     
     
         45 . The immunogenic composition of  claim 42  or  claim 43 , wherein the hemagglutinin proteins are derived from one or more interpandemic influenza virus strains. 
     
     
         46 . The immunogenic composition of  claim 42  or  claim 43 , wherein the hemagglutinin proteins are derived from a combination of one or more pandemic or potentially pandemic influenza virus strains and one or more interpandemic influenza virus strains.