Plant-derived vaccines against respiratory syncytial virus

Abstract

A plant-derived vaccine against respiratory syncytial virus (RSV) is disclosed. The vaccine includes an immunogenic complex that includes plant cells transformed with a chimeric gene containing a nucleotide sequence adapted for protein expression in plants and an RSV coding sequence that encodes an antigenic protein of RSV. Also disclosed are methods of making the plant-derived vaccine of the invention, as well as transgenic plants, transgenic plant cells, and nucleic acid constructs useful in immunizing a mammal against RSV.

1 - 22 . (canceled)  
     
     
         23 . A chimeric nucleic acid construct for expression in plants comprising: 
 a nucleotide sequence adapted for protein expression in plants comprising a respiratory syncytial virus (RSV) coding sequence encoding RSV-F protein or an antigenic RSV-F protein or peptide;    wherein the RSV-F coding sequence expresses in an edible portion of the plant, and    wherein upon oral ingestion of the edible portion of the plant by a subject in need thereof, the RSV-F coding sequence predominantly elicits a Th1 immune response and stimulates the mucosal immune system in the subject.    
     
     
         24 . The chimeric nucleic acid construct according to  claim 23 , wherein said nucleotide sequence includes a plant functional promoter.  
     
     
         25 . The chimeric nucleic acid construct according to  claim 24 , further comprising: 
 a translation enhancer;    wherein said translation enhancer is fused between said plant functional promoter and said RSV coding sequence.    
     
     
         26 . The chimeric nucleic acid construct according to  claim 25 , further comprising: 
 a transcriptional enhancer;    wherein said transcriptional enhancer is fused to the plant functional promoter.    
     
     
         27 . The chimeric nucleic acid construct according to  claim 24  wherein the plant functional promoter is an E8 promoter or a CaMV 35 S promoter.  
     
     
         28 . The chimeric nucleic acid construct according to  claim 25  wherein the translational enhancer is AMV RNA4 untranslated leader sequence.  
     
     
         29 . The chimeric nucleic acid construct according to  claim 26  wherein the transcriptional enhancer is a P268 enhancer fused upstream of the plant functional promoter.  
     
     
         30 . The chimeric nucleic acid construct according to  claim 23 , wherein the chimeric nucleic acid construct encodes a protein antigenically related to native RSV-F protein.  
     
     
         31 . The chimeric nucleic acid construct according to  claim 23 , wherein the nucleotide sequence comprises codons optimized for enhanced expression of RSV-F in plants.  
     
     
         32 . The nucleic acid construct according to  claim 23 , wherein said RSV coding sequence encodes an RSV-F glycoprotein or an antigenic RSV-F protein or peptide of RSV-F glycoprotein.  
     
     
         33 . A plant cell transformed with the chimeric nucleic acid construct according to  claim 23  conferring RSV-F antigen production in the plant cell.  
     
     
         34 . A transgenic plant transformed with the chimeric nucleic acid construct according to  claim 23  conferring RSV-F antigen production in the transgenic plant.  
     
     
         35 . The transgenic plant of  claim 34  wherein the plant is a tomato or apple plant.  
     
     
         36 . The transgenic plant of  claim 35  wherein the plant is a tomato plant.  
     
     
         37 . A method for preparing an immunogenic complex comprising: 
 growing plant cells transformed with the chimeric nucleic acid construct according to  claim 23;  and    expressing said RSV-F coding sequence in said plant cells to confer RSV-F antigen production in the plant cells.    
     
     
         38 . A method for preparing an immunogenic complex comprising: 
 transforming a plant with the chimeric nucleic acid construct according to  claim 23;  and    expressing the RSV-F coding sequence in the plant to confer RSV-F antigen production in the plant.    
     
     
         39 . An immunogenic complex comprising plant cells transformed with the chimeric nucleic acid construct according to  claim 23 .  
     
     
         40 . An immunogenic complex comprising a transgenic plant transformed with the chimeric nucleic acid construct according to  claim 23 .  
     
     
         41 . A method for inhibiting and/or treating RSV comprising: 
 administering to a subject in need thereof an antigen to RSV-F derived from a plant,    said antigen obtained from a plant transformed with a nucleic acid construct comprising a nucleotide sequence adapted for protein expression in plants comprising a respiratory syncytial virus (RSV) coding sequence encoding RSV-F protein or an antigenic RSV-F protein or peptide;    wherein the RSV-F coding sequence expresses in an edible portion of the plant, and    wherein upon oral ingestion of the edible portion of the plant by a subject in need thereof, the RSV-F coding sequence predominantly elicits a Th1 immune response and stimulates the mucosal immune system in the subject.