Influenza virus-like particles (vlps) comprising hemagglutinin produced within a plant

Abstract

A method for synthesizing influenza virus-like particles (VLPs) within a plant or a portion of a plant is provided. The method involves expression of influenza HA in plants and the purification by size exclusion chromatography. The invention is also directed towards a VLP comprising influenza HA protein and plants lipids. The invention is also directed to a nucleic acid encoding influenza HA as well as vectors. The VLPs may be used to formulate influenza vaccines, or may be used to enrich existing vaccines.

1 . A nucleic acid comprising a nucleotide sequence encoding an influenza hemagglutinin (HA) operatively linked to a regulatory region active in a plant. 
     
     
         2 . The nucleic acid of  claim 1 , wherein the influenza HA is selected from the group consisting of H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15, and H16. 
     
     
         3 . A method of producing influenza virus like particles (VLPs) in a plant comprising:
 a) introducing the nucleic acid of  claim 1  into the plant, or portion thereof, and   b) incubating the plant under conditions that permit the expression of the nucleic acid, thereby producing the VLPs.   
     
     
         4 . The method of  claim 3 , wherein in the step of introducing (step a), the nucleic acid is transiently expressed in the plant. 
     
     
         5 . The method of  claim 3 , wherein, in the step of introducing (step a), the nucleic acid is stably expressed in the plant. 
     
     
         6 . The method of  claim 3  further comprising a step of
 c) harvesting the host and purifying the VLPs. 
 
     
     
         7 . A virus like particle (VLP) produced by the method of  claim 3  comprising an influenza virus hemagglutinin (HA) protein and one or more than one lipid derived from a plant. 
     
     
         8 . The virus like particle (VLP) of  claim 7 , wherein the influenza HA protein is H5 Indonesia. 
     
     
         9 . A composition comprising an effective dose of the VLP of  claim 7  for inducing an immune response and a pharmaceutically acceptable carrier. 
     
     
         10 . A method of inducing immunity to an influenza virus infection in a subject, comprising administering the virus like particle of  claim 7 . 
     
     
         11 . The method of  claim 10 , wherein the virus like particle is administered to a subject orally, intradermally, intranasally, intramuscularly, intraperitoneally, intravenously, or subcutaneously. 
     
     
         12 . A virus like particle (VLP) produced by the method of  claim 3  comprising an influenza virus HA bearing plant-specific N-glycans, or modified N-glycans. 
     
     
         13 . A composition comprising an effective dose of the VLP of  claim 12  for inducing an immune response and a pharmaceutically acceptable carrier. 
     
     
         14 . A method of inducing immunity to an influenza virus infection in a subject, comprising administering the composition of  claim 13 . 
     
     
         15 . The method of  claim 11 , wherein the composition is administered to a subject orally, intradermally, intranasally, intramuscularly, intraperitoneally, intravenously, or subcutaneously.