PRRSV GP5 BASED COMPOSITIONS AND METHODS
The disclosure includes compositions and methods for the production of an immune response against porcine reproductive and respiratory syndrome (PRRS) virus, or PRRSV. The disclosure is based in part on the use of two or more peptide domains, each with a different sequence, from the PRRSV GP5 protein ectodomain. Compositions and methods comprising polypeptides containing the two or more domains, or nucleic acids encoding them, are described.
1 . An immunogenic composition, comprising:
a first separated PRRSV GP5 antigenic polypeptide comprising a first HV-2 hypervariable region; and
a second separated PRRSV GP5 antigenic polypeptide comprising a second HV-2 hypervariable region, the second HV-2 region selected to be different from the first HV-2 hypervariable region.
2 . The composition of claim 1 , wherein the first HV-2 hypervariable region has a first group sequence, and the second HV-2 hypervariable region has a second group sequence, selected to be different from the first group sequence.
3 . The composition of claim 2 , wherein the first HV-2 hypervariable region comprises a first sequence having an amino acid sequence X 0 WLX 1 X 2 X 3 X 4 X 5 , and the first group sequence is one of D1 to E-8, and the second HV-2 hypervariable region comprises a second sequence having an amino acid sequence X 0 WLX 1 X 2 X 3 X 4 X 5 , and the second group sequence is selected from D-1 to E-8; wherein the group sequences D-1 to E-8 are:
D-1: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is glycine, alanine, valine, leucine or isoleucine and X 2 is aspartic acid, asparagine, glutamic acid or glutamine, or X 1 is aspartic acid, asparagine, glutamic acid or glutamine and X 2 is glycine, alanine, valine, leucine or isoleucine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is glycine, alanine, valine, leucine, isoleucine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, lysine or histidine;
D-2: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is glycine, alanine, valine, leucine or isoleucine and X 2 is serine, threonine, tyrosine, arginine, lysine or histidine, or X 1 is serine, threonine, tyrosine, arginine, lysine or histidine and X 2 is glycine, alanine, valine, leucine or isoleucine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is glycine, alanine, valine, leucine, isoleucine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, lysine or histidine;
D-3: X 0 is one of the 20 naturally occurring amino acid residues; each of X 1 and X 2 is independently glycine, alanine, valine, leucine or isoleucine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is glycine, alanine, valine, leucine, isoleucine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, lysine or histidine;
D-4: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is aspartic acid, asparagine, glutamic acid or glutamine and X 2 is serine, threonine, tyrosine, arginine, lysine or histidine, or X 1 is serine, threonine, tyrosine, arginine, lysine or histidine and X 2 is aspartic acid, asparagine, glutamic acid or glutamine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is glycine, alanine, valine, leucine, isoleucine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, lysine or histidine;
D-5: X 0 is one of the 20 naturally occurring amino acid residues; each of X 1 and X 2 is independently aspartic acid, asparagine, glutamic acid or glutamine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is glycine, alanine, valine, leucine, isoleucine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, lysine or histidine;
D-6: X 0 is one of the 20 naturally occurring amino acid residues; each of X 1 and X 2 is independently one of the 20 naturally occurring amino acid residues; X 3 is aspartic acid, asparagine, glutamic acid or glutamine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is glycine, alanine, valine, leucine, isoleucine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, lysine or histidine;
D-7: X 0 is one of the 20 naturally occurring amino acid residues; each of X 1 and X 2 is independently one of the 20 naturally occurring amino acid residues; X 3 is serine, X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is glycine, alanine, valine, leucine, isoleucine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, lysine or histidine;
D-8: X 0 is one of the 20 naturally occurring amino acid residues; each of X 1 and X 2 is independently serine, threonine, tyrosine, arginine, lysine or histidine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is glycine, alanine, valine, leucine, isoleucine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, lysine or histidine;
S-1: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is aspartic acid, asparagine, glutamic acid or glutamine; X 2 is asparagine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is serine;
S-2: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is aspartic acid, asparagine, glutamic acid or glutamine; X 2 is aspartic acid, glutamic acid or glutamine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is serine;
S-3: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is glycine, alanine, valine, leucine or isoleucine and X 2 is aspartic acid, asparagine, glutamic acid or glutamine, or X 1 is aspartic acid, asparagine, glutamic acid or glutamine and X 2 is glycine, alanine, valine, leucine or isoleucine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is serine;
S-4: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is glycine, alanine, valine, leucine or isoleucine and X 2 is serine, threonine, tyrosine, arginine, lysine or histidine, or X 1 is serine, threonine, tyrosine, arginine, lysine or histidine and X 2 is glycine, alanine, valine, leucine or isoleucine; or each of X 1 and X 2 is independently serine, threonine, tyrosine, arginine, lysine or histidine, or each of X 1 and X 2 is independently glycine, alanine, valine, leucine or isoleucine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is serine;
S-5: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is aspartic acid, asparagine, glutamic acid or glutamine and X 2 is serine, threonine, tyrosine, arginine, lysine or histidine, or X 1 is serine, threonine, tyrosine, arginine, lysine or histidine and X 2 is aspartic acid, glutamic acid or glutamine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is serine;
S-6: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is serine, threonine, tyrosine, arginine, lysine or histidine; X 2 is asparagine; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is serine;
S-7: X 0 is one of the 20 naturally occurring amino acid residues; each of X 1 and X 2 is independently one of the 20 naturally occurring amino acid residues; X 3 is arginine, lysine or histidine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is threonine or tyrosine;
S-8: X 0 is one of the 20 naturally occurring amino acid residues; X 1 is aspartic acid, asparagine, glutamic acid or glutamine and X 2 is aspartic acid, asparagine, glutamic acid or glutamine, or X 1 is aspartic acid, asparagine, glutamic acid or glutamine and X 2 is glycine, alanine, valine, leucine or isoleucine, or X 1 is glycine, alanine, valine, leucine or isoleucine and X 2 is aspartic acid, asparagine, glutamic acid or glutamine; X 3 is aspartic acid, asparagine, glutamic acid or glutamine; X 4 is phenylalanine, tyrosine or tryptophan; and X 5 is serine;
E1: X 0 is asparagine; X 1 is serine; each of X 2 , X 3 , and X 4 is independently one of the 20 naturally occurring amino acid residues; and X 5 is aspartic acid, asparagine, glutamic acid, glutamine, glycine, alanine, valine, leucine or isoleucine;
E2: X 0 is aspartic acid, glutamic acid or glutamine; each of X 1 , X 2 , X 3 , and X 4 is independently one of the 20 naturally occurring amino acid residues; and X 5 is aspartic acid, asparagine, glutamic acid, glutamine, glycine, alanine, valine, leucine or isoleucine;
E3: X 0 is arginine, lysine or histidine; each of X 1 , X 2 , X 3 , and X 4 is independently one of the 20 naturally occurring amino acid residues; and X 5 is aspartic acid, asparagine, glutamic acid, glutamine, glycine, alanine, valine, leucine or isoleucine;
E4: X 0 is glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, phenylalanine, tyrosine, tryptophan or proline; each of X 1 , X 2 , X 3 , and X 4 is independently one of the 20 naturally occurring amino acid residues; and X 5 is aspartic acid, asparagine, glutamic acid, glutamine, glycine, alanine, valine, leucine or isoleucine;
E5: X 0 is asparagine; X 1 is serine; each of X 2 , X 3 and X 4 is independently one of the 20 naturally occurring amino acid residues; and X 5 is serine or threonine;
E6: X 0 is any of the 20 naturally occurring amino acid residues except asparagine; X 2 is asparagine; each of X 1 , X 3 , and X 4 is independently one of the 20 naturally occurring amino acid residues; and X 5 is serine or threonine;
E7: X 0 is aspartic acid, glutamic acid or glutamine; each of X 1 , X 2 , X 3 , and X 4 is independently one of the 20 naturally occurring amino acid residues except that X 2 is not asparagine; and X 5 is glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, arginine, lysine, histidine, phenylalanine, tyrosine, tryptophan or proline; and
E8: X 0 is glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, arginine, lysine, histidine, phenylalanine, tyrosine, tryptophan or proline; each of X 1 , X 2 , X 3 , and X 4 is independently one of the 20 naturally occurring amino acid residues; and X 5 is glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, arginine, lysine, histidine, phenylalanine, tyrosine, tryptophan or proline.
4 . The composition of claim 1 , further comprising an adjuvant.
5 . The composition of claim 3 , wherein at least one of the first and second group sequences is D-1, D-2, D-3, D-4, D-5, D-6, D-7 or D-8.
6 . The composition of claim 3 , wherein at least one of the first and second group sequences is S-1, S-2, S-3, S-4, S-5, S-6, S-7 or S-8.
7 . The composition of claim 3 , wherein at least one of the first and second group sequences is E-1, E-2, E-3, E-4, E-5, E-6, E-7 or E-8.
8 . The composition of claim 3 , wherein the first group sequence is D-1, D-2, D-3, D-4, D-5, D-6, D-7 or D-8, and the second group sequence is S-1, S-2, S-3, S-4, S-5, S-6, S-7 or S-8.
9 . The composition of claim 1 , further comprising a third PRRSV GP5 antigenic polypeptide comprising a third HV-2 hypervariable region.
10 . The composition of claim 9 , wherein the third HV-2 hypervariable region is selected to be different from the first and second HV-2 hypervariable regions.
11 . The composition of claim 9 , wherein the third PRRSV GP5 antigenic polypeptide is a separated PRRSV GP5 antigenic polypeptide.
12 . The composition of claim 9 , wherein the first HV-2 hypervariable region has a first group sequence, the second HV-2 hypervariable region has a second group sequence different from the first group sequence, and the third HV-2 hypervariable region has a third group sequence different from the first and second group sequences.
13 . The composition of claim 1 , comprising isolated first and second PRRSV GP5 antigenic polypeptides.
14 . The composition of claim 1 , wherein each of the first and second PRRSV GP5 antigenic polypeptides independently has a length from 14 to 201 amino acids.
15 . An immunogenic composition, comprising a first GP5 antigenic polypeptide domain having a length from 14 to 201 amino acids comprising a first HV-2 hypervariable region having a length of about 8 amino acids, and a second GP5 antigenic polypeptide domain having a length from 14 to 201 amino acids comprising a second HV-2 hypervariable region having a length of about 8 amino acids, the second HV-2 region selected to be different from the first HV-2 hypervariable region, the immunogenic composition being substantially free of intact viral particles.
16 . A method of making an immunogenic composition, comprising:
identifying or selecting a first PRRSV GP5 antigenic polypeptide comprising a first HV-2 hypervariable region;
purposefully selecting a second PRRSV GP5 antigenic polypeptide that has a second HV-2 hypervariable region different from the first HV-2 hypervariable region; and
combining the first and second PRRSV GP5 antigenic polypeptides to form the immunogenic composition.
17 . The method of claim 16 , wherein the first HV-2 hypervariable region has a first group sequence, and the second HV-2 hypervariable region has a second group sequence.
18 . The method of claim 17 , comprising purposefully selecting the second PRRSV GP5 antigenic polypeptide such that the second group sequence is different from the first group sequence.
19 . The method of claim 16 , further comprising adding an adjuvant.
20 . The method of claim 16 , further comprising combining a third PRRSV GP5 antigenic polypeptide with the first and second PRRSV GP5 antigenic polypeptides.
21 . The method of claim 20 , comprising selecting the third PRRSV GP5 antigenic polypeptide to have a third HV-2 hypervariable region that is different from the first and second HV-2 hypervariable regions.
22 . The method of claim 21 , wherein the third HV-2 hypervariable region has a third group sequence, and selecting the third GP5 antigenic polypeptide comprises selecting the third group sequence such that it is different from at least one of the first and second group sequences.
23 . The method of claim 22 , wherein the third group sequence is different from both the first and second group sequences.
24 . The method of claim 16 , wherein the first PRRSV GP5 antigenic polypeptide is a first separated PRRSV GP5 antigenic polypeptide, and the second PRRSV GP5 antigenic polypeptide is a second separated PRRSV GP5 antigenic polypeptide.
25 . An immunogenic composition made by the method of claim 16 .