Recombinant swine influenza virus and uses thereof

Abstract

Recombinant, chimeric porcine influenza viruses are disclosed that include hemagglutinin segments from more than one influenza virus subtype. Also described are methods of producing the recombinant influenza viruses, immunogenic compositions comprising the recombinant influenza viruses, methods of stimulating an immune response against influenza virus, and methods of treating and preventing influenza virus infection.

1 . A recombinant, chimeric porcine influenza virus comprising more than one hemagglutinin (HA) segment (segment 5) from more than one influenza subtype, wherein said virus comprises segments 1-5, 7 and 8 from a first influenza subtype and a second segment 5 from a second influenza subtype, and further wherein all or a portion of the neuraminidase (NA) segment (segment 6) of the first influenza subtype is missing to render an attenuated virus. 
     
     
         2 . The recombinant, chimeric porcine influenza virus of  claim 1 , wherein said second segment 5 comprises NA packaging sequences from said first influenza subtype located 3′ and optionally 5′ to said second segment 5. 
     
     
         3 . The recombinant, chimeric porcine influenza virus of  claim 2 , wherein the NA packaging sequences comprise 3′ NA packaging sequences from the 3′ NA UTR and the 3′ NA coding sequence and, optionally 5′ NA packaging sequences from the 5′ NA UTR and the 5′ NA coding sequence. 
     
     
         4 . The recombinant, chimeric porcine influenza virus of  claim 1 , wherein the influenza virus is derived from an influenza A virus. 
     
     
         5 . The recombinant, chimeric porcine influenza virus of  claim 4 , wherein the influenza virus comprises an HA segment from an H1N1 subtype and an HA segment from an H3N2 subtype. 
     
     
         6 . The recombinant, chimeric porcine influenza virus of  claim 1 , wherein the first influenza subtype is H1N1. 
     
     
         7 . The recombinant, chimeric porcine influenza virus of  claim 6 , wherein the H1N1 subtype is A/swine/Saskatchewan/18789/02. 
     
     
         8 . The recombinant, chimeric porcine influenza virus of  claim 1 , wherein the second influenza subtype is H3N2. 
     
     
         9 . The recombinant, chimeric porcine influenza virus of  claim 8 , wherein the H3N2 subtype is A/Swine/Texas/4199-2/98. 
     
     
         10 . An attenuated, recombinant, porcine influenza virus comprising segments 1-5, 7 and 8 from an H1N1 influenza subtype, and segment 5 from an H3N2 influenza subtype, wherein all or a portion of segment 6 from the H1N1 influenza subtype is missing, wherein the H3N2 segment 5 is flanked by NA packaging sequences from said H1N1 subtype, wherein the packaging sequences comprise 3′ NA packaging sequences from the 3′ NA UTR and the 3′ NA coding sequence and 5′ NA packaging sequences from the 5′ NA UTR and the 5′ NA coding sequence. 
     
     
         11 . The attenuated, recombinant porcine influenza virus of  claim 10 , wherein the H1N1 subtype is A/swine/Saskatchewan/18789/02 and the H3N2 subtype is A/Swine/Texas/4199-2/98. 
     
     
         12 . A composition comprising the recombinant virus of  claim 1  and a pharmaceutically acceptable excipient. 
     
     
         13 . The composition of  claim 12 , further comprising an adjuvant. 
     
     
         14 . A composition comprising the recombinant virus of  claim 10  and a pharmaceutically acceptable excipient. 
     
     
         15 . The composition of  claim 14 , further comprising an adjuvant. 
     
     
         16 . A method of eliciting an immunological response in a vertebrate subject, comprising administering the composition of  claim 12  to said subject. 
     
     
         17 . A method of eliciting an immunological response in a vertebrate subject, comprising administering the composition of  claim 14  to said subject. 
     
     
         18 . A method of treating or preventing an influenza infection in a vertebrate subject, comprising administering to said subject a therapeutically effective amount of the composition of  claim 14 . 
     
     
         19 . A method of vaccinating a subject against an influenza virus, comprising administering an effective amount of the composition of  claim 14  to said subject. 
     
     
         20 . The method of  claim 16 , wherein the subject is a porcine subject. 
     
     
         21 . The method of  claim 20 , wherein the influenza virus is a swine influenza virus. 
     
     
         22 . A recombinant construct comprising:
 (a) a porcine influenza H3N2 subtype HA segment; and   (b) porcine influenza H1N1 subtype NA packaging sequences located 3′ and optionally 5′ to said H3N2 HA segment.   
     
     
         23 . The recombinant construct of  claim 22 , wherein the H3N2 HA segment is flanked by H1N1 NA packaging sequences that comprise 3′ NA packaging sequences from the 3′ NA UTR and the 3′ NA coding sequence and 5′ NA packaging sequences from the 5′ NA UTR and the 5′ NA coding sequence. 
     
     
         24 . The recombinant construct of  claim 23 , wherein the H1N1 subtype is A/swine/Saskatchewan/18789/02 and the H3N2 subtype is A/Swine/Texas/4199-2/98. 
     
     
         25 . A method of producing a recombinant, chimeric influenza virus, comprising transfecting a host cell with (a) individual plasmids comprising segments 1-5, 7 and 8 from an H1N1 influenza subtype; and (b) a recombinant construct according to  claim 22 , and culturing said host cell under conditions that result in the production of said recombinant, chimeric influenza virus. 
     
     
         26 . A cell transformed with (a) individual plasmids comprising segments 1-5, 7 and 8 from an H1N1 influenza subtype; and (b) a recombinant construct according to  claim 22 . 
     
     
         27 . A method of producing a composition comprising combining the recombinant, chimeric porcine influenza virus of  claim 1  with a pharmaceutically acceptable excipient. 
     
     
         28 . A method of producing a composition comprising combining the recombinant, chimeric porcine influenza virus of  claim 10  with a pharmaceutically acceptable excipient. 
     
     
         29 . A method of producing an influenza vaccine comprising:
 (a) propagating the recombinant, chimeric porcine influenza virus of  claim 1 ;   (b) purifying the virus; and   (c) combining the purified virus with a pharmaceutically acceptable excipient.   
     
     
         30 . A method of producing an influenza vaccine comprising:
 (a) propagating the recombinant, chimeric porcine influenza virus of  claim 10 ;   (b) purifying the virus; and   (c) combining the purified virus with a pharmaceutically acceptable excipient.   
     
     
         31 . A kit comprising one or more containers wherein the one or more containers comprise the recombinant virus of  claim 1 .