Expression systems

Abstract

The invention relates to an expression system comprising polynucleotides encoding proteins, wherein the expression system comprises a first polynucleotide encoding at least one protein, peptide or variant thereof, which induces a T cell response, and a second polynucleotide encoding at least one protein, peptide or variant thereof, which induces an anti-pathogenic B cell response. The invention further relates to protein mixtures encoded by the expression system and cells comprising the expression system or the protein mixture and pharmaceutical compositions comprising the expression system or the protein mixture.

1 . An expression system comprising polynucleotides encoding proteins, wherein the expression system comprises a first polynucleotide encoding at least one protein, peptide or variant thereof, which induces a T cell response, and a second polynucleotide encoding at least one protein peptide or variant thereof, which induces an anti-pathogenic B cell response. 
     
     
         2 . The expression system of  claim 1  wherein the first polynucleotide and the second polynucleotide are linked such that they are expressed as an artificial polyprotein. 
     
     
         3 . The expression system of  claim 1 , wherein the proteins, peptides or fragments encoded by the first and the second polynucleotide are separated co- or posttranslationally. 
     
     
         4 . The expression system of  claim 1 , wherein a polynucleotide which encodes a cleavage site is positioned between the first polynucleotide and the second polynucleotide. 
     
     
         5 . The expression system of  claim 4 , wherein the cleavage site is a self-cleaving site or an endopeptidase cleavage site. 
     
     
         6 . (canceled) 
     
     
         7 . The expression-system of  claim 1 , further comprising a third polynucleotide encoding a protein or variant thereof. 
     
     
         8 .- 13 . (canceled) 
     
     
         14 . The expression system of  claim 1 , wherein the expression system comprises at least one vector selected from the group consisting of plasmid vectors, cosmid vectors, phage vectors, and viral vectors, viral like particles, and bacterial spores. 
     
     
         15 . The expression-system of  claim 14 , wherein the viral vector is an adenovirus vector selected from the group consisting of PanAd1, PanAd2, PanAd3, ChAd55, ChAd 73, ChAd83, ChAd146, ChAd147, ChAd3, ChAd4, ChAd5, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd16, ChAd17, ChAd19, ChAd20, ChAd22, ChAd24, ChAd26, ChAd30, ChAd31, ChAd37, ChAd38, ChAd44, ChAd63 and ChAd82. 
     
     
         16 . The expression-system of  claim 14 , wherein the viral vector is selected from the group consisting of cytomegaloviruse vectors, arena virus vectors, and pox virus vectors. 
     
     
         17 . The expression system of  claim 1 , wherein at least one of the first polynucleotide or the second polynucleotide is selected from the group consisting of a protein, peptide or variant thereof from a pathogen. 
     
     
         18 . The expression system of  claim 17 , wherein the pathogen is a virus. 
     
     
         19 . The expression system of  claim 1 , wherein the protein, that induces a T cell response is a non-structural and/or internal protein of a virus, and/or the protein that induces an anti-pathogenic B cell response is a structural and/or surface protein of a virus. 
     
     
         20 .- 23 . (canceled) 
     
     
         24 . The expression system of  claim 18 , wherein the virus is selected from the group consisting of paramyxoviruses and orthomyxoviruses. 
     
     
         25 .- 26 . (canceled) 
     
     
         27 . The expression system of  claim 24 , wherein the paramyxovirus is selected from the group consisting of Pneumovirus, Metapneumovirus, and Respirovirus. 
     
     
         28 . (canceled) 
     
     
         29 . The expression system of  claim 19 , wherein the structural and/or surface protein of the virus is selected from the group of paramyxovirus proteins consisting of fusion protein (F), and any of the attachment glycoproteins G, H, and HN. 
     
     
         30 .- 31 . (canceled) 
     
     
         32 . The expression system of  claim 1 , wherein the non-structural and/or internal protein is selected from the group of paramyxovirus proteins consisting of nucleoprotein N, Matrix proteins M and M2, Phosphoprotein P, non structural proteins NS1 and NS2, and the catalytic subunit of the polymerase (L). 
     
     
         33 .- 35 . (canceled) 
     
     
         36 . The expression system of  claim 24 , wherein the orthomyxovirus is selected from Influenza A virus, Influenza B virus, Influenza C virus, Thogotovirus, Isavirus and unclassified Orthomyxoviridae. 
     
     
         37 . (canceled) 
     
     
         38 . The expression system of  claim 19 , wherein the structural and/or surface protein of the virus is selected from the group orthomyxovirus proteins consisting of hemagglutinin (HA) and neuraminidase (NA). 
     
     
         39 .- 40 . (canceled) 
     
     
         41 . The expression system of  claim 19 , wherein the non-structural and/or internal protein(s) is selected from the group of orthomyxovirus proteins consisting of nucleoprotein (NP), matrixprotein 1 (MP1), matrixprotein M2, non-structural protein 1 (NS1), non-structural protein2/nuclear export protein (NS2/NEP), polymerase subunit protein PA, polymerase subunit protein PB1, polymerase subunit protein PB2, and PB1-F2 protein encoded by an alternate reading frame in the PB1 gene. 
     
     
         42 .- 43 . (canceled) 
     
     
         44 . An isolated protein mixture encoded by the expression system of  claim 1 . 
     
     
         45 . An isolated host cell containing the expression-system according to  claim 1 . 
     
     
         46 . A composition comprising the expression-system according to  claim 1  and a pharmaceutically acceptable carrier and/or excipient. 
     
     
         47 .- 50 . (canceled) 
     
     
         51 . A method of treatment or prevention of a viral disease comprising administration of an effective amount of the expression-system according to  claim 1 . 
     
     
         52 . The method of  claim 51  wherein the disease is caused by a virus selected from the group consisting of cytomegalovirus, arena virus, pox virus, paramyxovirus, and orthomyxovirus. 
     
     
         53 . (canceled) 
     
     
         54 . A nucleic acid construct which comprises a polynucleotide encoding a modified influenza hemagglutinin (HA), wherein the HA0 cleavage site is modified by introducing one or more basic amino acids. 
     
     
         55 .- 57 . (canceled) 
     
     
         58 . The expression system of  claim 1 , comprising at least one polynucleotide encoding a modified influenza HA protein, wherein the HA0 cleavage site is modified by introducing one or more basic amino acids. 
     
     
         59 . An isolated protein mixture encoded by the expression system of  claim 58 . 
     
     
         60 . An isolated host cell containing the expression-system according to  claim 58 . 
     
     
         61 . A composition comprising the expression-system of  claim 58  and a pharmaceutically acceptable carrier and/or excipient. 
     
     
         62 .- 63 . (canceled) 
     
     
         64 . A method of treatment or prevention of a viral disease comprising administration of an effective amount of the expression-system according to claim  586 . 
     
     
         65 . (canceled)