Peptides for vaccine against birch allergy

Abstract

The present invention relates to compositions comprising peptides for preventing or treating allergy to birch, and in particular to optimal combinations of peptides for preventing or treating said allergy.

1 . A composition of matter suitable for use in preventing or treating allergy to birch pollen, said composition being selected from the group consisting of:
 a) a composition comprising:
 i) at least one of the polypeptides of SEQ ID NO: 74 (BIR12B; AKYMVIQGEPGRVIRGK), SEQ ID NO: 72 (BIR11; FPQFKPQEITGIMK), SEQ ID NO: 71 (BIR10; GSVWAQSSSFPQFK), SEQ ID NO: 73 (BIR12A; PTGMFVAGAKYMVIQGR), SEQ ID NO: 75 (BIR13; IKYMVIQGEAGAVIRGK) and SEQ ID NO: 76 (BIR14; EAGAVIRGKKGSGGIT), or a variant of any thereof, and 
 ii) at least one of the polypeptides of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK), SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK), SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV), SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY), SEQ ID NO: 60 (BIR07; SNEIKIVATPDGGSILK), and SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK), or a variant of any thereof, 
   wherein a variant of a polypeptide is:
 I) up to 30 amino acids in length and comprises the sequence of the said polypeptide, or 
 II) 9 to 30 amino acids in length and comprises a sequence that has at least 65% homology to the sequence of said polypeptide, which sequence is capable of tolerising to said polypeptide; or 
 III) 9 to 30 amino acids in length and comprises a sequence of at least 9 contiguous amino acids of the sequence of said polypeptide, or a sequence that has at least 65% homology to said at least 9 contiguous amino acids, which sequence of at least 9 contiguous amino acids or homologous sequence is capable of tolerising to said polypeptide; 
   b) a composition comprising at least one polynucleotide sequence which when expressed causes the production of a composition as defined in (i);   c) a vector comprising four or more polynucleotide sequences which each encode a different polypeptide as defined in (i);   d) a product comprising:
 i) at least one of the polypeptides of SEQ ID NO: 74 (BIR12B; AKYMVIQGEPGRVIRGK), SEQ ID NO: 72 (BIR11; FPQFKPQEITGIMK), SEQ ID NO: 71 (BIR10; GSVWAQSSSFPQFK), SEQ ID NO: 73 (BIR12A; PTGMFVAGAKYMVIQGR), SEQ ID NO: 75 (BIR13; IKYMVIQGEAGAVIRGK) and SEQ ID NO: 76 (BIR14; EAGAVIRGKKGSGGIT), or a variant of any thereof, and 
   ii) at least one of the polypeptides of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK), SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK), SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV), SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY), SEQ ID NO: 60 (BIR07; SNEIKIVATPDGGSILK), and SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK), or a variant of any thereof,   wherein a variant of a polypeptide is as defined in  claim 1 (a) (I), (II) or (III), and   wherein each different polypeptide is for simultaneous, separate or sequential use in preventing or treating allergy to birch pollen; and   e) a pharmaceutical formulation comprising a composition according to (a) or (b); a vector according to (c); or a product according to (d); and a pharmaceutically acceptable carrier or diluent, and optionally one or more adjuvants selected from a glucocorticoid, vitamin D and rapamycin.   
     
     
         2 . A composition according to  claim 1 (a), which is selected from the group consisting of:
 i) a composition of  claim 1 (a) which further comprises at least one additional polypeptide of  claim 1 (a)(i) or (ii) or variant thereof not selected in  claim 1 (a);   ii) a composition of  claim 1 (a) which further comprises at least one additional polypeptide selected from the group consisting of SEQ ID NO: 77 (BIR15; SLNTLRLRRIFDLFDK) and SEQ ID NO:78 (BIR16A; AERERIFKRFDANGEGK), or a variant of any thereof; and   iii) a composition of  claim 1 (a) which comprises:   (a) the polypeptide SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK), or a variant thereof;   (b) the polypeptide SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK), or a variant thereof; and   (c) the polypeptide SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a variant thereof;   
       wherein a variant of a polypeptide is as defined in  claim 1 (a) (I), (II) or (III), preferably wherein;
 when said polypeptide is SEQ ID NO: 48 said variant of SEQ ID NO: 48 is SEQ ID NO: 45 (Bir01F; FNYETEATSVIPAARK), SEQ ID NO: 46 (Bir01G; FNYEIEATSVIPAARK) or SEQ ID NO: 47 (Bir01H; FNYEIETTSVIPAARK); and 
 when said polypeptide is SEQ ID NO: 53, said variant of SEQ ID NO: 53 is SEQ ID NO: 50 (Bir02E; PAARLFKAFILEGDTLIPK), SEQ ID NO: 51 (Bir02G; PAARLFKAFILEGDNLIPK), SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or SEQ ID NO: 49 (Bir02D; PAARMFKAFILDGDKLVPK); and 
 when said polypeptide is SEQ ID NO: 67, said variant of SEQ ID NO: 67 is selected from SEQ ID NO: 68 (Bir09A; GETLLRAVESYLLAHS), SEQ ID NO: 69 (Bir09B; KEMGETLLRAVESYLLAHS) or SEQ ID NO: 70 (Bir09C; KEKGETLLRAVESYLLAHS); and 
 when said polypeptide is SEQ ID NO: 78 said variant of SEQ ID NO: 78 is SEQ ID NO: 79 (Bir16B; AERERIFKRFDAGGEGK). 
 
     
     
         3 - 5 . (canceled) 
     
     
         6 . A composition suitable for use in preventing or treating allergy to birch pollen comprising at least three different polypeptides, selected from:
 (a) SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK), or a variant thereof;   (b) SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK), or a variant thereof;   (c) SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a variant thereof;   (d) SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a variant thereof;   (e) SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY) or a variant thereof;   (f) SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a variant thereof;   (g) SEQ ID NO: 60 (Bir07; SNEIKIVATPDGGSILK) or a variant thereof;   (h) SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a variant thereof;   (i) SEQ ID NO: 72 (Bir01I; FPQFKPQEITGIMK) or a variant thereof;   (j) SEQ ID NO: 77 (Bir15; SLNTLRLRRIFDLFDK) or a variant thereof;   
       wherein a variant of a polypeptide is: 
       I) up to 30 amino acids in length and comprises the sequence of said polypeptide, or 
       II) 9 to 30 amino acids in length and comprises a sequence that has at least 65% homology to the sequence of said polypeptide, which sequence is capable of tolerising to said polypeptide; or 
       III) 9 to 30 amino acids in length and comprises a sequence of at least 9 contiguous amino acids of the sequence of said polypeptide, or a sequence that has at least 65% homology to said at least 9 contiguous amino acids, which sequence of at least 9 contiguous amino acids or homologous sequence is capable of tolerising to said polypeptide. 
     
     
         7 . A composition according to  claim 6 , which is selected from the group consisting of:
 i) a composition comprising:   a) the polypeptide SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a variant thereof;   b) at least one of the polypeptides SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) and SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK), or a variant of any thereof; and   c) at least one additional polypeptide of a) to j);   ii) a composition comprising the polypeptide SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a variant thereof, the polypeptide SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a variant thereof, the polypeptide SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a variant thereof, and at least one additional polypeptide of a) to j);   iii) a composition comprising the polypeptide SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a variant thereof, the polypeptide SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a variant thereof, the polypeptide SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a variant thereof, the polypeptide SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a variant thereof, the polypeptide SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY) or a variant thereof, the polypeptide SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a variant thereof, and the polypeptide SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a variant thereof, and optionally no further polypeptides derived from a birch pollen allergen;   iv) a composition comprising the polypeptide SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a variant thereof, the polypeptide SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a variant thereof, the polypeptide SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a variant thereof, the polypeptide SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a variant thereof, the polypeptide SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a variant thereof, the polypeptide SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a variant thereof, and the polypeptide SEQ ID NO: 69 (Bir09B; KEMGETLLRAVESYLLAHS), and optionally no further polypeptides derived from a birch pollen allergen; and   v) a composition comprising the polypeptide SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a variant thereof, the polypeptide SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a variant thereof, the polypeptide SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a variant thereof, the polypeptide SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a variant thereof, the polypeptide SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a variant thereof, and the polypeptide SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a variant thereof, and optionally no further polypeptides derived from a birch pollen allergen;   
       wherein a variant of a polypeptide is as defined in claim  3  (I), (II) or (III), preferably wherein:
 when said polypeptide is SEQ ID NO: 48 said variant of SEQ ID NO: 48 is SEQ ID NO: 45 (Bir01F; FNYETEATSVIPAARK), SEQ ID NO: 46 (Bir01G; FNYEIEATSVIPAARK) or SEQ ID NO: 47 (Bir01H; FNYEIETTSVIPAARK); and 
 when said polypeptide is SEQ ID NO: 53, said variant of SEQ ID NO: 53 is SEQ ID NO: 50 (Bir02E; PAARLFKAFILEGDTLIPK), SEQ ID NO: 51 (Bir02G; PAARLFKAFILEGDNLIPK), SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or SEQ ID NO: 49 (Bir02D; PAARMFKAFILDGDKLVPK); and 
 when said polypeptide is SEQ ID NO: 67, said variant of SEQ ID NO: 67 is selected from SEQ ID NO: 68 (Bir09A; GETLLRAVESYLLAHS), SEQ ID NO: 69 (Bir09B; KEMGETLLRAVESYLLAHS) or SEQ ID NO: 70 (Bir09C; KEKGETLLRAVESYLLAHS); and 
 when said polypeptide is SEQ ID NO: 78, said variant of SEQ ID NO: 78 is SEQ ID NO: 79 (Bir 16B; AERERIFKRFDAGGEGK). 
 
     
     
         8 - 12 . (canceled) 
     
     
         13 . The composition according to  claim 1 (a), wherein the composition:
 is capable of tolerising at least 50% or at least 60% of a panel of birch pollen allergic individuals in the population and/or   comprises at least one further polypeptide up to a total of thirteen unique/different polypeptides, wherein the further polypeptides:   
       (a) comprise a sequence having at least 65% sequence identity to at least 9 or more contiguous amino acids in any of SEQ ID NOs: 1 to 80 not selected above; and 
       (b) are 9 to 30 amino acids in length. 
     
     
         14 . The composition according to  claim 1 (a), comprising at least one said polypeptide which is 9 to 20 or 13 to 17 amino acids in length and/or wherein said polypeptide has at least 70% sequence identity to any of SEQ ID NOS: 1 to 80. 
     
     
         15 . The composition according to  claim 1 (a), wherein one or more of the polypeptides have one or more modifications selected from the following:
 (i) N terminal acetylation;   (ii) C terminal amidation;   (iii) one or more hydrogens on the side chain amines of Arginine and/or Lysine replaced with a methylene group;   (iv) glycosylation; and   (v) phosphorylation.   
     
     
         16 . The composition according to  claim 1 (a) wherein at least one of the polypeptides has been engineered to be soluble such that it comprises:
 i) N terminal to the residues of the polypeptide which flank a T cell epitope: one to six contiguous amino acids corresponding to the two to six contiguous amino acids immediately N terminal to said residues in the sequence of the protein from which the polypeptide derives; and/or   ii) C terminal to the residues of the polypeptide which flank a T cell epitope: one to six contiguous amino acids corresponding to the one to six contiguous amino acids immediately C terminal to the said residues in the sequence of the protein from which the polypeptide derives; or   iii) both N and C terminal to the residues of the polypeptide which flank a T cell epitope, at least one amino acid selected from arginine, lysine, histidine, glutamate and aspartate, wherein the polypeptide has a solubility of at least 3.5 mg/ml and the T cell epitope has a solubility of less than 3.5 mg/ml.   
     
     
         17 . The composition according to  claim 1 (a) wherein at least one of the polypeptides has been engineered to be soluble such that additionally:
 i) any cysteine residues in the native sequence of the polypeptide are replaced with serine or 2-aminobutyric acid; and/or   ii) any hydrophobic residues in the up to three amino acids at the N or C terminus of the native sequence of the polypeptide, which are not comprised in a T cell epitope, are deleted; and/or   iii) any two consecutive amino acids comprising the sequence Asp-Gly in the up to four amino acids at the N or C terminus of the native sequence of the polypeptide, which are not comprised in a T cell epitope, are deleted; and/or   iv) one or more positively charged residues are added at the N and/or C terminus of the native sequence of the polypeptide.   
     
     
         18 . The composition according to  claim 1 (a) wherein each polypeptide has a concentration in the range of 0.03 to 200 nmol/ml, 0.3 to 200 nmol/ml, 50 to 200 nmol/ml or 30 to 120 nmol/ml. 
     
     
         19 - 23 . (canceled) 
     
     
         24 . The composition according to  claim 1 (e), wherein said composition of  claim 1 (a) or (b), said vector of  claim 1 (c) or said product of  claim 1 (d) is formulated for oral administration, nasal administration, topical administration, subcutaneous administration, sublingual administration, intradermal administration, buccal administration, epidermal administration, or for administration by inhalation, by injection, or by a patch. 
     
     
         25 . The composition according to  claim 1  wherein said composition of  claim 1 (a) or product of  claim 1 (d), additionally comprises a further polypeptide allergen for use in tolerising an individual to the further polypeptide allergen. 
     
     
         26 . An in vitro method of determining whether T cells recognize a composition according to  claim 1 (a), comprising contacting said T cells with said composition and detecting whether said T cells are stimulated by said composition. 
     
     
         27 . A method according to  claim 26  which is carried out to determine whether an individual has, or is at risk of having, an allergy to birch pollen. 
     
     
         28 . A method of preventing or treating allergy to birch pollen, the method comprising administering to a subject in need of such treatment a therapeutically effective amount of:
 (A) a composition comprising:
 i) at least one of the polypeptides of SEQ ID NO: 74 (BIR12B; AKYMVIQGEPGRVIRGK), SEQ ID NO: 72 (BIR11; FPQFKPQEITGIMK), SEQ ID NO: 71 (BIR10; GSVWAQSSSFPQFK), SEQ ID NO: 73 (BIR12A; PTGMFVAGAKYMVIQGR), SEQ ID NO: 75 (BIR13; IKYMVIQGEAGAVIRGK) and SEQ ID NO: 76 (BIR14; EAGAVIRGKKGSGGIT), or a variant of any thereof, and 
 ii) at least one of the polypeptides of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK), SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK), SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV), SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY), SEQ ID NO: 60 (BIR07; SNEIKIVATPDGGSILK), and SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK), or a variant of any thereof, 
   wherein a variant of a polypeptide is:
 I) up to 30 amino acids in length and comprises the sequence of the said polypeptide, or 
 II) 9 to 30 amino acids in length and comprises a sequence that has at least 65% homology to the sequence of said polypeptide, which sequence is capable of tolerising to said polypeptide; or 
 III) 9 to 30 amino acids in length and comprises a sequence of at least 9 contiguous amino acids of the sequence of said polypeptide, or a sequence that has at least 65% homology to said at least 9 contiguous amino acids, which sequence of at least 9 contiguous amino acids or homologous sequence is capable of tolerising to said polypeptide. 
   or   (B) a composition comprising at least three different polypeptides, selected from:
 (a) SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK), or a variant thereof; 
 (b) SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK), or a variant thereof; 
 (c) SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a variant thereof; 
 (d) SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a variant thereof; 
 (e) SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY) or a variant thereof; 
 (f) SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a variant thereof; 
 (g) SEQ ID NO: 60 (Bir07; SNEIKIVATPDGGSILK) or a variant thereof; 
 (h) SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a variant thereof; 
 (i) SEQ ID NO: 72 (Bir011; FPQFKPQEITGIMK) or a variant thereof; 
 (j) SEQ ID NO: 77 (Bir15; SLNTLRLRRIFDLFDK) or a variant thereof; 
   wherein a variant of a polypeptide is:
 I) up to 30 amino acids in length and comprises the sequence of said polypeptide, or 
 II) 9 to 30 amino acids in length and comprises a sequence that has at least 65% homology to the sequence of said polypeptide, which sequence is capable of tolerising to said polypeptide; or 
 III) 9 to 30 amino acids in length and comprises a sequence of at least 9 contiguous amino acids of the sequence of said polypeptide, or a sequence that has at least 65% homology to said at least 9 contiguous amino acids, which sequence of at least 9 contiguous amino acids or homologous sequence is capable of tolerising to said polypeptide. 
   
     
     
         29 . A composition according to  claim 1 , which is selected from the group consisting of:
 i) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a said variant thereof, the polypeptide of SEQ ID NO:48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof; and optionally no further polypeptides derived from a birch pollen allergen;   ii) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof, and the polypeptide of SEQ ID NO: 69 (Bir09B; KEMGETLLRAVESYLLAHS) or a said variant thereof, and optionally no further polypeptides derived from a birch pollen allergen;   iii) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof, and optionally no further polypeptides derived from a birch pollen allergen;   iv) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof;   v) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof, and the polypeptide of SEQ ID NO: 69 (Bir09B; KEMGETLLRAVESYLLAHS) or a said variant thereof; and   vi) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK).   
     
     
         30 . The method according to  claim 28 , wherein the composition is selected from the group consisting of:
 i) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a said variant thereof, the polypeptide of SEQ ID NO:48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof; and optionally no further polypeptides derived from a birch pollen allergen;   ii) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof, and the polypeptide of SEQ ID NO: 69 (Bir09B; KEMGETLLRAVESYLLAHS) or a said variant thereof, and optionally no further polypeptides derived from a birch pollen allergen;   iii) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof, and optionally no further polypeptides derived from a birch pollen allergen;   iv) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof;   v) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or said variant thereof, and the polypeptide of SEQ ID NO: 69 (Bir09B; KEMGETLLRAVESYLLAHS) or a said variant thereof; and   vi) a composition comprising the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK).   
     
     
         31 . A method of preparing a pharmaceutical formulation, the method comprising combining with a pharmaceutically acceptable carrier or diluent
 i) at least one of the polypeptides of SEQ ID NO: 74 (BIR12B; AKYMVIQGEPGRVIRGK), SEQ ID NO: 72 (BIR11; FPQFKPQEITGIMK), SEQ ID NO: 71 (BIR10; GSVWAQSSSFPQFK), SEQ ID NO: 73 (BIR12A; PTGMFVAGAKYMVIQGR), SEQ ID NO: 75 (BIR13; IKYMVIQGEAGAVIRGK and SEQ ID NO: 76 (BIR14; EAGAVIRGKKGSGGIT), or a variant of any thereof, and   ii) at least one of the polypeptides of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK), SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK), SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV), SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY), SEQ ID NO: 60 (BIR07; SNEIKIVATPDGGSILK), and SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK), or a variant of any thereof,   
       wherein a variant of a polypeptide is:
 I) up to 30 amino acids in length and comprises the sequence of the said polypeptide, or 
 II) 9 to 30 amino acids in length and comprises a sequence that has at least 65% homology to the sequence of said polypeptide, which sequence is capable of tolerising to said polypeptide; or 
 III) 9 to 30 amino acids in length and comprises a sequence of at least 9 contiguous amino acids of the sequence of said polypeptide, or a sequence that has at least 65% homology to said at least 9 contiguous amino acids, which sequence of at least 9 contiguous amino acids or homologous sequence is capable of tolerising to said polypeptide. 
 
     
     
         32 . A method according to  claim 31 , the method comprising combining with a pharmaceutically acceptable carrier or diluent
 i) the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a said variant thereof, the polypeptide of SEQ ID NO:48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof; and optionally no further polypeptides derived from a birch pollen allergen;   ii) the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof, and the polypeptide of SEQ ID NO: 69 (Bir09B; KEMGETLLRAVESYLLAHS) or a said variant thereof, and optionally no further polypeptides derived from a birch pollen allergen;   iii) the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 53 (Bir02J; PAARMFKAFILEGDKLVPK) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof, and optionally no further polypeptides derived from a birch pollen allergen;   iv) the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 67 (Bir09; ETLLRAVESYLLAHSDAY) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof;   v) the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK) or a said variant thereof, and the polypeptide of SEQ ID NO: 69 (Bir09B; KEMGETLLRAVESYLLAHS) or a said variant thereof; or   vi) the polypeptide of SEQ ID NO: 74 (Bir12B; AKYMVIQGEPGRVIRGK) or a said variant thereof, the polypeptide of SEQ ID NO: 52 (Bir02I; PAARMFKAFILD) or a said variant thereof, the polypeptide of SEQ ID NO: 48 (Bir01I; FNYETETTSVIPAARK) or a said variant thereof, the polypeptide of SEQ ID NO: 54 (Bir04; PGTIKKISFPEGFPFKYV) or a said variant thereof, the polypeptide of SEQ ID NO: 63 (Bir07C; SNEIKIVATPEGGSILK) or a said variant thereof, and the polypeptide of SEQ ID NO: 78 (Bir16A; AERERIFKRFDANGEGK).