Dengue virus e-glycoprotein polypeptides containing mutations that eliminate immunodominant cross-reactive epitopes

Abstract

Described herein are dengue virus E-glycoprotein polypeptides containing mutations that eliminate immunodominant cross-reactive epitopes associated with immune enhancement. The disclosed dengue virus E-glycoproteins optionally further include mutations that introduce a strong CD4 T cell epitope. The disclosed E-glycoprotein polypeptides, or nucleic acid molecules encoding the polypeptides, can be used, for example, in monovalent or tetravalent vaccines against dengue virus.

1 . An isolated cross-reactivity reduced dengue virus E-glycoprotein polypeptide, wherein the polypeptide is selected from:
 (1) a dengue serotype 1 virus (DENV-1) E-glycoprotein polypeptide comprising an arginine at position 106, an aspartic acid at position 107, an aspartic acid at position 310, a lysine at position 311, a glutamine at position 364, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487, numbered with reference to SEQ ID NO: 1;   (2) a dengue serotype 2 virus (DENV-2) E-glycoprotein polypeptide comprising an arginine at position 106, an aspartic acid at position 107, an glutamic acid at position 310, an arginine at position 311, an arginine at position 364, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487, numbered with reference to SEQ ID NO: 2;   (3) a dengue serotype 3 virus (DENV-3) E-glycoprotein polypeptide comprising an aspartic acid at position 106, an aspartic acid at position 107, an aspartic acid at position 308, a lysine at position 309, an aspartic acid at position 362, an isoleucine at position 466, a threonine at position 476, a valine at position 480 and a leucine at position 485, numbered with reference to SEQ ID NO: 3; and   (4) a dengue serotype 4 virus (DENV-4) E-glycoprotein polypeptide comprising a glutamic acid at position 106, an aspartic acid at position 107, an aspartic acid at position 310, a glutamine at position 311, a glutamine at position 389, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487, numbered with reference to SEQ ID NO: 4.   
     
     
         2 . The polypeptide of  claim 1 , wherein the amino acid sequence of the polypeptide is at least 95% identical to SEQ ID NO: 1, and wherein the polypeptide comprises an arginine at position 106, an aspartic acid at position 107, an aspartic acid at position 310, a lysine at position 311, a glutamine at position 364, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487 of SEQ ID NO: 1. 
     
     
         3 . (canceled) 
     
     
         4 . The polypeptide of  claim 1 , wherein the amino acid sequence of the polypeptide is at least 95% identical to SEQ ID NO: 2, and wherein the polypeptide comprises an arginine at position 106, an aspartic acid at position 107, an glutamic acid at position 310, an arginine at position 311, an arginine at position 364, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487 of SEQ ID NO: 2. 
     
     
         5 . (canceled) 
     
     
         6 . The polypeptide of  claim 1 , wherein the amino acid sequence of the polypeptide is at least 95% identical to SEQ ID NO: 3, and wherein the polypeptide comprises an aspartic acid at position 106, an aspartic acid at position 107, an aspartic acid at position 308, a lysine at position 309, an aspartic acid at position 362, an isoleucine at position 466, a threonine at position 476, a valine at position 480 and a leucine at position 485 of SEQ ID NO: 3. 
     
     
         7 . (canceled) 
     
     
         8 . The polypeptide of  claim 1 , wherein the amino acid sequence of the polypeptide is at least 95% identical to SEQ ID NO: 4, and wherein the polypeptide comprises a glutamic acid at position 106, an aspartic acid at position 107, an aspartic acid at position 310, a glutamine at position 311, a glutamine at position 389, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487 of SEQ ID NO: 4. 
     
     
         9 . The polypeptide of  claim 1 , wherein the amino acid sequence of the polypeptide comprises SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. 
     
     
         10 . An isolated dengue virus-like particle (VLP) comprising the polypeptide of  claim 1 . 
     
     
         11 . The VLP of  claim 10 , further comprising a dengue virus prM protein. 
     
     
         12 . A recombinant nucleic acid molecule encoding the polypeptide of  claim 1 . 
     
     
         13 . The recombinant nucleic acid molecule of  claim 12 , comprising a nucleotide sequence at least 95%, identical to SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 12. 
     
     
         14 . The recombinant nucleic acid molecule of  claim 12 , comprising the nucleotide sequence of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 12. 
     
     
         15 . A vector comprising the recombinant nucleic acid molecule of  claim 12 . 
     
     
         16 . An isolated cell comprising the vector of  claim 15 . 
     
     
         17 . A composition comprising the polypeptide of  claim 1 , and a pharmaceutically acceptable carrier. 
     
     
         18 . The composition of  claim 17 , further comprising an adjuvant. 
     
     
         19 . A tetravalent dengue virus vaccine, comprising a recombinant nucleic acid molecule encoding the DENV-1 polypeptide of SEQ ID NO: 1, a recombinant nucleic acid molecule encoding the DENV-2 polypeptide of SEQ ID NO: 2, a recombinant nucleic acid molecule encoding the DENV-3 polypeptide of SEQ ID NO: 3, and a recombinant nucleic acid molecule encoding the DENV-4 polypeptide of SEQ ID NO: 4. 
     
     
         20 . A method of eliciting an immune response in a subject against dengue virus, comprising administering to the subject a therapeutically effective amount of the vaccine of  claim 19 , thereby eliciting an immune response in the subject against dengue virus. 
     
     
         21 - 22 . (canceled) 
     
     
         23 . A composition comprising a DENV-1 E-glycoprotein polypeptide, a DENV-2 E-glycoprotein polypeptide, a DENV-3 E-glycoprotein polypeptide and a DENV-4 E-glycoprotein polypeptide; or a recombinant nucleic acid molecule encoding the DENV-1 E-glycoprotein polypeptide, a recombinant nucleic acid molecule encoding the DENV-2 E-glycoprotein polypeptide, a recombinant nucleic acid molecule encoding the DENV-3 E-glycoprotein polypeptide and a recombinant nucleic acid molecule encoding the DENV-4 E-glycoprotein polypeptide, wherein:
 (1) the DENV-1 E-glycoprotein polypeptide comprising an arginine at position 106, an aspartic acid at position 107, an aspartic acid at position 310, a lysine at position 311, a glutamine at position 364, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487, numbered with reference to SEQ ID NO: 1;   (2) the DENV-2 E-glycoprotein polypeptide comprising an arginine at position 106, an aspartic acid at position 107, an glutamic acid at position 310, an arginine at position 311, an arginine at position 364, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487, numbered with reference to SEQ ID NO: 2;   (3) the DENV-3 E-glycoprotein polypeptide comprising an aspartic acid at position 106, an aspartic acid at position 107, an aspartic acid at position 308, a lysine at position 309, an aspartic acid at position 362, an isoleucine at position 466, a threonine at position 476, a valine at position 480 and a leucine at position 485, numbered with reference to SEQ ID NO: 3; and   (4) the DENV-4 E-glycoprotein polypeptide comprising a glutamic acid at position 106, an aspartic acid at position 107, an aspartic acid at position 310, a glutamine at position 311, a glutamine at position 389, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487, numbered with reference to SEQ ID NO: 4.   
     
     
         24 . The composition of  claim 23 , wherein:
 (1) the amino acid sequence of the DENV-1 polypeptide is at least 95% identical to SEQ ID NO: 1, and wherein the polypeptide comprises an arginine at position 106, an aspartic acid at position 107, an aspartic acid at position 310, a lysine at position 311, a glutamine at position 364, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487 of SEQ ID NO: 1;   (2) the amino acid sequence of the DENV-2 polypeptide is at least 95% identical to SEQ ID NO: 2, and wherein the polypeptide comprises an arginine at position 106, an aspartic acid at position 107, an glutamic acid at position 310, an arginine at position 311, an arginine at position 364, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487 of SEQ ID NO: 2;   (3) the amino acid sequence of the DENV-3 polypeptide is at least 95% identical to SEQ ID NO: 3, and wherein the polypeptide comprises an aspartic acid at position 106, an aspartic acid at position 107, an aspartic acid at position 308, a lysine at position 309, an aspartic acid at position 362, an isoleucine at position 466, a threonine at position 476, a valine at position 480 and a leucine at position 485 of SEQ ID NO: 3; and   (4) the amino acid sequence of the DENV-4 polypeptide is at least 95% identical to SEQ ID NO: 4, and wherein the polypeptide comprises a glutamic acid at position 106, an aspartic acid at position 107, an aspartic acid at position 310, a glutamine at position 311, a glutamine at position 389, an isoleucine at position 468, a threonine at position 478, a valine at position 482 and a leucine at position 487 of SEQ ID NO: 4.   
     
     
         25 . The composition of  claim 23 , wherein the DENV-1 polypeptide comprises the amino acid sequence of SEQ ID NO: 1, the DENV-2 polypeptide comprises the amino acid sequence of SEQ ID NO: 2, the DENV-3 polypeptide comprises the amino acid sequence of SEQ ID NO: 3 and the DENV-4 polypeptide comprises the amino acid sequence of SEQ ID NO: 4. 
     
     
         26 . A method of eliciting an immune response in a subject against dengue virus, comprising administering to the subject a therapeutically effective amount of the composition of  claim 23 . 
     
     
         27 . The method of  claim 26 , further comprising administering a protein boost vaccination. 
     
     
         28 . A method of eliciting an immune response in a subject against dengue virus, comprising administering to the subject a therapeutically effective amount of the polypeptide of  claim 1 , thereby eliciting an immune response in the subject against dengue virus.