CAT Allergen Conjugates and Uses Thereof

Abstract

The present invention is in the fields of medicine, public health, immunology, molecular biology and virology. The invention provides compositions comprising a virus-like particle (VLP) or a virus particle and at least one antigen, particularly at least one feline antigen, and more particularly at least one feline antigen that is a human allergen. In certain embodiments, the antigen is a Fel d1 antigen or a fragment thereof, covalently linked to the VLP. The invention also provides methods for producing the compositions. The compositions of the invention induce efficient immune responses, in particular antibody responses, in mammals, particularly humans. The compositions and methods of the invention are useful in the production of vaccines, in particular for the treatment and/or prevention of allergies to cat dander and other cat antigens and allergens.

What is claimed is: 
     
         1 . A composition comprising:
 (a) a core particle with at least one first attachment site, wherein said core particle is a virus-like particle (VLP) or a virus particle,   (b) at least one antigen with at least one second attachment site, wherein said at least one antigen is a Fel d1 protein or a Fel d1 fragment, and wherein (a) and (b) are covalently linked through said at least one first and said at least one second attachment site.   
     
     
         2 . The composition of  claim 1 , wherein said Fel d1 protein comprises chain 1 of Fel d1 and chain 2 of Fel d1, wherein said chain 1 of Fel d1 is associated with chain 2 of Fel d1 by at least one covalent bond. 
     
     
         3 . The composition of  claim 1  or  2 , wherein said Fel d1 protein is a fusion protein comprising chain 1 of Fel d1 and chain 2 of Fel d1, wherein chain 1 of Fel d1 and chain 2 of Fel d1 are fused either directly via one peptide bond or via a spacer, which links the N-terminus of one chain with the C-terminus of another chain. 
     
     
         4 . The composition of  claim 3 , wherein said chain 2 of Fel d1 is fused via its C-terminus to the N-terminus of said chain 1 of Fel d1 either directly or via a spacer. 
     
     
         5 . The composition of  claim 3 , wherein said chain 1 of Fel d1 is fused via its C-tenninus to the N-terminus of said chain 2 of Fel d1 either directly or via a spacer. 
     
     
         6 . The composition of any one of the  claims 3 - 5 , wherein said spacer consists of an amino acid sequence having 1-20 amino acid residues. 
     
     
         7 . The composition of any one of the  claims 3 - 6 , wherein said fusion protein comprises an amino acid sequence selected from the group consisting of:
 (a) SEQ ID NO: 24;   (b) SEQ ID NO:54;   (c) SEQ ID NO:55;   (d) SEQ ID NO:56; and   (e) SEQ ID NO:57.   
     
     
         8 . The composition of any one of the  claims 3 - 6 , wherein said chain 1 of Fel d 1 comprises a sequence of SEQ ID NO: 22 or a homologue sequence thereof, wherein said homologue sequence has an identity to SEQ ID NO: 22 of greater than 80%, preferably greater than 90%, or even more preferably greater than 95%. 
     
     
         9 . The composition of any one of the  claims 3 - 6 , wherein said chain 2 of Fel d 1 comprises a sequence of SEQ ID NO: 23, SEQ ID NO: 25 or SEQ ID NO: 26, or a homologue sequence thereof, wherein said homologue sequence has an identity to SEQ ID NO: 23, SEQ ID NO: 25 or SEQ ID NO: 26 of greater than 80%, preferably greater than 90%, and even more preferably greater than 95%. 
     
     
         10 . The composition of any one of the proceeding claims, wherein said core particle is a virus particle, and wherein preferably said virus particle is a virus particle of an RNA-bacteriophage. 
     
     
         11 . The composition of any one of the  claims 1 - 9 , wherein said core particle is a virus-like particle, and wherein preferably said virus-like particle is a virus-like particle of an RNA-bacteriophage. 
     
     
         12 . The composition of  claim 10  or  11 , wherein said RNA-bacteriophage is selected from Qβ, fr, GA or AP205. 
     
     
         13 . The composition of any of the proceeding claims, wherein said first attachment site comprises, or preferably is, an amino group, preferably an amino group of a lysine. 
     
     
         14 . The composition of any of the proceeding claims, wherein said second attachment site comprises, or preferably is, a sulfhydryl group, preferably a sulfhydryl group of a cysteine. 
     
     
         15 . The composition of any of the proceeding claims further comprising a linker. 
     
     
         16 . The composition of  claim 15 , wherein said linker is fused to the C-terminus of said Fel d1 protein or to the C-terminus of said Fel d1 fragment. 
     
     
         17 . A vaccine comprising the composition of any one of the  claims 1 - 16 . 
     
     
         18 . The vaccine of  claim 17  further comprising at least one adjuvant. 
     
     
         19 . A pharmaceutical composition comprising:
 (a) the composition of any of the  claims 1 - 16 ; and   (b) an acceptable pharmaceutical carrier.   
     
     
         20 . A method of producing the composition of any one of the  claims 1 - 16  comprising:
 (a) providing a core particle with at least one first attachment site, wherein said core particle is a virus-like particle (VLP) or a virus particle; 
 (b) providing at least one antigen with at least one second attachment site, wherein said antigen is a Fel d1 protein or a Fel d1 fragment; and 
 (c) linking said core particle and said at least one antigen to produce said composition, wherein said at least one antigen and said core particle are linked through said at least one first and said at least one second attachment sites. 
 
     
     
         21 . Use of the composition of any one of the  claims 1 - 16  for the manufacture of a medicament for the treatment of cat allergy. 
     
     
         22 . A method of treating cat allergy comprising administering the composition of any one of the  claims 1 - 16  or the vaccine of any one of the  claims 17 - 18  to a human or to a non-human mammal, wherein preferably said non-human mammal is a dog or a cat. 
     
     
         23 . The composition of any one of the  claims 1 - 16  or the vaccine of any one of the  claims 17 - 18  for use in a method for treating cat allergy comprising administering said composition or said vaccine to a human or to a non-human mammal, wherein preferably said non-human mammal is a dog or a cat. 
     
     
         24 . A Fel d1 fusion protein comprising chain 1 of Fel d1 and chain 2 of Fel d1 fused via an amino acid spacer, which links the N-terminus of one chain with the C-terminus of another chain, wherein said amino acid spacer consists of an amino acid sequence having 10-30 amino acid residues, and wherein said fusion protein is not a fusion protein comprising chain 1 of SEQ ID NO:22 fused through (GGGGS) 3  to the N-terminus of chain 2 of SEQ ID NO:23, 25 or 26 and wherein said disclaimed fusion protein is expressed in baculovirus expression system. 
     
     
         25 . The Fel d1 fusion protein of  claim 24 , wherein said spacer consists of an amino acid sequence having 2, 3 or 4 times of GGGGS repeat. 
     
     
         26 . The Fel d1 fusion protein of  claim 24  or  25 , wherein said Fel d1 fusion protein is produced in  E. coli.    
     
     
         27 . The Fel d1 fusion protein of any one of the  claims 24 - 26 , wherein chain 2 of Fel d1 is at the N-terminus of the fusion protein. 
     
     
         28 . The Fel d1 fusion protein of any one of the  claims 24 - 26 , wherein chain 1 of Fel d1 is at the N-terminus of the fusion protein. 
     
     
         29 . The Fel d1 fusion protein of any one of the  claims 24 - 28  comprising an amino acid sequence selected from the group consisting of:
 (a) SEQ ID NO:54; 
 (b) SEQ ID NO:55; and 
 (c) SEQ ID NO:57. 
 
     
     
         30 . A Fel d1 fusion protein comprising an amino acid sequence of SEQ ID NO:57, wherein said Fel d1 protein is produced in  E. coli.