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 . A method of classifying a PRRSV isolate, the method comprising sequencing the HV-2 hypervariable region of a PRRSV isolate, and identifying or classifying the PRRSV isolate based upon the HV-2 hypervariable region.
2 . The method of claim 1 , wherein the HV-2 hypervariable region is of about eight amino acid residues that follow a conserved GP5 motif, represented by the sequence C(E/S)LNG(T/A), SEQ ID NO:1.
3 . The method of claim 1 , wherein the HV-2 hypervariable region is represented by the sequence X 0 WLX 1 X 2 X 3 X 4 X 5 , wherein each of X 0 , X 1 , X 2 , X 3 , X 4 , and X 5 is independently one of the 20 naturally occurring amino acid residues, except that X 5 is not cysteine, methionine, proline, phenylalanine, or tryptophan.
4 . The method of claim 3 , wherein the HV-2 region comprises the sequence X 0 WLX 1 X 2 X 3 X 4 X 5 wherein X 5 is not cysteine, methionine, proline, phenylalanine, tryptophan, serine, threonine, or tyrosine, and is identified or classified as
D-1: wherein X 1 is an aliphatic amino acid residue, X 2 is an acidic amino acid residue (or wherein X 1 is an acidic amino acid residue and X 2 is an aliphatic amino acid residue), X 3 is a basic amino acid residue, and X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F);
D-2: wherein X 1 is an aliphatic amino acid residue, X 2 is serine (S), threonine (T), tyrosine (Y) or an basic amino acid residue (or wherein X 1 is serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue and X 2 is an aliphatic amino acid residue), X 3 is a basic amino acid residue, and X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F);
D-3: wherein each of X 1 and X 2 is independently an aliphatic amino acid residue, X 3 is a basic amino acid residue, and X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F);
D-4: wherein X 1 is an acidic amino acid residue, X 2 is serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue (or wherein X 1 is serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue and X 2 is an acidic amino acid residue), X 3 is a basic amino acid residue, and X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F);
D-5: wherein each of X 1 and X 2 is independently an acidic amino acid residue, X 3 is a basic amino acid residue, and X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F);
D-6: wherein each of X 1 and X 2 is independently one of the 20 naturally occurring amino acid residues, X 3 is an acidic amino acid residue, and X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F);
D-7: wherein each of X 1 and X 2 is independently one of the 20 naturally occurring amino acid residues, X 3 is a non-aromatic amino acid residue with a hydroxyl containing R-group, and X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F); or
D-8: wherein each of X 1 and X 2 is independently serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue, X 3 is a basic amino acid residue, and X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F).
5 . The method of claim 1 , wherein the HV-2 region comprises the sequence X 0 WLX 1 X 2 X 3 X 4 X 5 and is identified or classified as
S-1: wherein X 1 is an acidic amino acid residue, X 2 is asparagine (N), X 3 is a basic amino acid residue, X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F), and X 5 is serine (S);
S-2: wherein each of X 1 and X 2 is independently an acidic amino acid residue except that X 2 is not asparagine (N), X 3 is a basic amino acid residue, X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F), and X 5 is serine (S);
S-3: wherein X 1 is an aliphatic amino acid residue, X 2 is an acidic amino acid residue (or wherein X 1 is an acidic amino acid residue and X 2 is an aliphatic amino acid residue), X 3 is a basic amino acid residue, X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F), and X 5 is serine (S);
S-4: wherein X 1 is an aliphatic amino acid residue, X 2 is serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue (or wherein X 1 is serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue and X 2 is an aliphatic amino acid residue; or where each of X 1 and X 2 is independently serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue; or where each of X 1 and X 2 is independently an aliphatic amino acid residue), X 3 is a basic amino acid residue, X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F), and X 5 is serine (S);
S-5: wherein X 1 is an acidic amino acid residue, X 2 is serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue (or wherein X 1 is serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue and X 2 is an acidic amino acid residue except N (Asn)), X 3 is a basic amino acid residue, X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F), and X 5 is serine (S);
S-6: wherein X 1 is serine (S), threonine (T), tyrosine (Y) or a basic amino acid residue, X 2 is an asparagine (N), X 3 is a basic amino acid residue, X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F), and X 5 is serine (S);
S-7: wherein each of X 1 and X 2 is independently one of the 20 naturally occurring amino acid residues, X 3 is a basic amino acid residue, X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F), and X 5 is threonine (T) or tyrosine (Y); or
S-8: wherein X 1 is an acidic amino acid residue, X 2 is an acidic amino acid residue (or wherein X 1 is an acidic amino acid residue and, X 2 is an aliphatic amino acid residue, or alternatively wherein X 1 is an aliphatic amino acid residue and, X 2 is an acidic amino acid residue), X 3 is an acidic amino acid residue, X 4 is an amino acid residue comprising an aromatic ring, such as phenylalanine (F), and X 5 is serine (S).
6 . The method of claim 1 , wherein the HV-2 region comprises
the sequence NWLSX 2 X 3 X 4 X 5 , wherein each of X 2 , X 3 , and X 4 is independently one of the 20 naturally occurring amino acid residues, and X 5 is an acidic amino acid residue, and is identified or classified as E-1;
the sequence X 0 WLX 1 X 2 X 3 X 4 X 5 , wherein X 0 is an acidic amino acid residue except for asparagine (N), 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 an acidic amino acid residue, and is identified or classified as E-2;
the sequence X 0 WLX 1 X 2 X 3 X 4 X 5 , wherein X 0 is a basic amino acid residue, 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 an acidic amino acid residue, and is identified or classified as E-3;
the sequence X 0 WLX 1 X 2 X 3 X 4 X 5 , wherein X 0 is any non-acidic and non-basic amino acid residue, 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 an acidic amino acid residue, and is identified or classified as E-4;
the sequence NWLSX 2 X 3 X 4 X 5 , wherein 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 (S) or Threonine (T), and is identified or classified as E-5;
the sequence X 0 WLX 1 NX 3 X 4 X 5 , wherein X 0 is any amino acid residue except asparagine (N), 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 (S) or Threonine (T), and is identified or classified as E-6;
the sequence X 0 WLX 1 X 2 X 3 X 4 X 5 , wherein X 0 is an acidic amino acid residue except asparagine (N), 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 (N), and X 5 is any non-acidic amino acid residue, and is identified or classified as E-7; or
the sequence X 0 WLX 1 X 2 X 3 X 4 X 5 , wherein X 0 is any non-acidic amino acid residue, 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 any non-acidic amino acid residue, and is identified or classified as E-8.
7 . The method of claim 1 , wherein the isolate is a PRRSV viral particle.
8 . The method of claim 7 , wherein the viral particle is a virion.
9 . The method of claim 1 , wherein the isolate is obtained from a pig.
10 . The method of claim 1 , wherein the sequencing is of the GP5 coding sequence (ORFS) of the isolate.
11 . The method of claim 1 , wherein the sequencing is of the GP5 protein.