Protein vaccines against poxviruses

Abstract

The invention described here entails a protein vaccine against poxviruses which contains at least two purified recombinant monkeypox virus proteins or peptides. The proteins or peptides are encoded by the open reading frames of the monkeypox ortholog genes M1R, A35R, A29L B6R, and orthologs of these proteins or peptides having 90% identity. The invention also entails a vaccine protocol against poxvirus whereby a vaccine is vaccinated with a first vaccine made up of a nucleic acid vaccine of three or more poxvirus virus genes, and subsequently vaccinated with at least one other booster vaccine made up of two or more poxvirus virus proteins.

1 . A vaccine against poxviruses comprising at least two purified recombinant monkeypox virus proteins or peptides selected from the group consisting of
 (i) a protein or peptide encoded by the open reading frame of the monkeypox ortholog M1R gene,   (ii) a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog M1R gene, which protein or peptide has 90% amino acid sequence identity to a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog M1R gene;   (iii) a protein or peptide encoded by the open reading frame of the monkeypox ortholog A29L gene,   (iv) a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog A29L gene, which protein or peptide has 90% amino acid sequence identity to a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog A29L gene;   (v) a protein or peptide encoded by the open reading frame of the monkeypox ortholog A35R gene,   (vi) a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog A35R gene, which protein or peptide has 90% amino acid sequence identity to a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog A35R gene;   (vii) a protein or peptide encoded by the open reading frame of the monkeypox ortholog B6R gene, and   (viii) a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog B6R gene, which protein or peptide has 90% amino acid sequence identity to a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog B6R gene;   wherein at least one protein or peptide is (i), (ii), (iii) or (iv) and at least one protein or peptide is (v), (vi), (vii) or (viii), and an adjuvant.   
     
     
         2 . The vaccine of  claim 1  wherein said poxvirus protected against is an orthopoxvirus selected from the group consisting of: camelpox virus, ectromelia virus, raccoon poxvirus, skunk poxvirus, Tatera poxvirus, Uasin Gishu virus, Volepox virus, variola virus, vaccinia virus, monkeypox virus, gerbilpox and cowpox virus, or genetically engineered versions thereof. 
     
     
         3 . The vaccine of  claim 1  wherein the proteins or peptides of the orthologs of (ii), (iv), (vi) and (viii) are derived from an orthopoxvirus selected from the group consisting of: camelpox virus, ectromelia virus, raccoon poxvirus, skunk poxvirus, Tatera poxvirus, Uasin Gishu virus, Volepox virus, variola virus, vaccinia virus, monkeypox virus, gerbilpox and cowpox virus, or genetically engineered versions thereof. 
     
     
         4 . The vaccine of  claim 1 , wherein the adjuvant is selected from the group consisting of CpG, alum, immune modulatory molecules, Toll-like receptor stimulators and co-stimulatory markers. 
     
     
         5 . The vaccine of  claim 1 , wherein the vaccine comprises a protein or peptide encoded by the open reading frame of the monkeypox ortholog M1R gene, a protein or peptide encoded by the open reading frame of the monkeypox ortholog A35R gene, a protein or peptide encoded by the open reading frame of the monkeypox ortholog A29L gene, a protein or peptide encoded by the open reading frame of the monkeypox ortholog B6R gene, and an adjuvant. 
     
     
         6 . The vaccine of  claim 1 , wherein the recombinant monkeypox virus proteins or peptides contain B cell epitopes. 
     
     
         7 . The vaccine of  claim 6  wherein the B cell epitope is selected from the group consisting of amino acids 49-64 or 237-263 within the protein or peptide encoded by the open reading frame of the monkeypox ortholog B6R gene, amino acids 97-127 within the a protein or peptide encoded by the open reading frame of the monkeypox ortholog A35R gene, and amino acids 69-91 or 137-155 within the protein or peptide encoded by the open reading frame of the monkeypox ortholog M1R gene. 
     
     
         8 . The vaccine of  claim 1 , wherein the purified recombinant monkeypox virus proteins or peptides are expressed from prokaryotic host cells or synthesized in vitro. 
     
     
         9 . The vaccine of  claim 8 , wherein the prokaryotic host cells are  E. coli  cells. 
     
     
         10 . A method for inducing in a subject an immune response against poxvirus infection comprising administering to said subject an immunologically effective amount of the vaccine of  claim 1 , in a pharmaceutically acceptable carrier. 
     
     
         11 . The method of  claim 10  for inducing in a subject an immune response against poxvirus, wherein the dosage amount is between 50 micrograms and 1 milligram. 
     
     
         12 . An immunogenic composition comprising at least two purified recombinant monkeypox virus proteins or peptides selected from the group consisting of
 (i) a protein or peptide encoded by the open reading frame of the monkeypox ortholog M1R gene,   (ii) a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog M1R gene, which protein or peptide has 90% amino acid sequence identity to a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog M1R gene;   (iii) a protein or peptide encoded by the open reading frame of the monkeypox ortholog A29L gene,   (iv) a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog A29L gene, which protein or peptide has 90% amino acid sequence identity to a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog A29L gene;   (v) a protein or peptide encoded by the open reading frame of the monkeypox ortholog A35R gene,   (vi) a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog A35R gene, which protein or peptide has 90% amino acid sequence identity to a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog A35R gene;   (vii) a protein or peptide encoded by the open reading frame of the monkeypox ortholog B6R gene, and   (viii) a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog B6R gene, which protein or peptide has 90% amino acid sequence identity to a protein or peptide encoded by the open reading frame of an ortholog of the monkeypox ortholog B6R gene;   wherein at least one protein or peptide is (i), (ii), (iii) or (iv) and at least one protein or peptide is (v), (vi), (vii) or (viii).   
     
     
         13 . The immunogenic composition of  claim 12  wherein the orthologs of (ii), (iv), (vi) and (viii) are derived from an orthopoxvirus selected from the group consisting of: camelpox virus, ectromelia virus, raccoon poxvirus, skunk poxvirus, Tatera poxvirus, Uasin Gishu virus, Volepox virus, variola virus, vaccinia virus, monkeypox virus, gerbilpox and cowpox virus, or genetically engineered versions thereof. 
     
     
         14 - 30 . (canceled)