Variants of group 5 allergens of the true grasses having reduced allergeneity due to mutagenesis of proline residues

Abstract

The present invention relates to the preparation and use of recombinant variants of group 5 allergens of the Poaceae (true grasses), which are characterised by reduced IgE reactivity compared with known wild-type allergens and at the same time substantially retained reactivity with T-lymphocytes.

1 . A hypoallergenic variant of a group 5 allergen of the true grass family (Poaceae) wherein the prolines which correspond in an alignment to the prolines in positions 57, 58, 117, 146, 155, 180, 211 in the amino acid sequence of wild-type Phl p 5.0109 have been mutated singly or in combinations. 
     
     
         2 . The hypoallergenic variant of  claim 1 , wherein the proline which corresponds in an alignment to proline 211 of wild-type Phl p 5.0109 is not deleted, but instead is replaced by another amino acid. 
     
     
         3 . The hypoallergenic variant of  claim 1 , wherein the prolines which correspond in an alignment to the prolines in positions 146 and 155 in the amino acid sequence of wild-type Phl p 5.0109 are not mutated. 
     
     
         4 . The hypoallergenic variant of  claim 1 , further wherein the amino acids which correspond in an alignment to lysine in position 61 and glutamic acid in position 205 in the amino acid sequence of wild-type Phl p 5.0109 have been mutated singly or in combinations. 
     
     
         5 . The hypoallergenic variant of  claim 1 , wherein the group 5 allergen is a group 5 allergen of the Pooideae sub-family. 
     
     
         6 . The hypoallergenic variant of  claim 1 , wherein the group 5 allergen is selected from the group consisting of Phl p 5, Lol p 5 , Poa  p 5, Hol l 5, Dac g 5, Pha a 5, Ant o 5, Fes p 5, Hor v 5, Sec c 5 and Tri a 5. 
     
     
         7 . The hypoallergenic variant of  claim 1 , wherein it is a fragment or a variant of a hypoallergenic variant of  claim 1  or a multimer of one or more hypoallergenic variants of  claim 1  or wherein one or more hypoallergenic variants of  claim 1  or fragments, variants or multimers thereof are a constituent of a recombinant fusion protein. 
     
     
         8 . A DNA molecule which encodes for a hypoallergenic variant of  claim 1 . 
     
     
         9 . A recombinant expression vector containing a DNA molecule of  claim 8  functionally connected to an expression control sequence. 
     
     
         10 . A non-human host organism transformed with a DNA molecule of  claim 8 . 
     
     
         11 . A process for the preparation of a hypoallergenic variant by cultivation of a non-human host organism of  claim 10  and isolation of the corresponding allergen variant from the culture. 
     
     
         12 . The hypoallergenic variant of  claim 1 , formulated as medicament. 
     
     
         13 . The DNA molecule of  claim 8 , formulated as medicament. 
     
     
         14 . A recombinant expression vector of  claim 9  as medicament. 
     
     
         15 . A method for the prevention and/or therapeutic treatment of type 1 allergies in the triggering of which group 5 allergens of the true grasses are causally involved, comprising administering a hypoallergenic variant of  claim 1 . 
     
     
         16 . A method for immunotherapeutic DNA vaccination comprising administering to a patient at least one DNA molecule of  claim 8 . 
     
     
         17 . Pharmaceutical preparation comprising at least one hypoallergenic variant of  claim 1  and optionally further active compounds and/or assistants for the prevention and/or therapeutic treatment of type 1 allergies. 
     
     
         18 . A pharmaceutical preparation of  claim 17 , wherein the further active compounds are allergens of the true grasses or variants thereof. 
     
     
         19 . A pharmaceutical preparation comprising at least one hypoallergenic variant of  claim 8  and optionally further active compounds and/or assistants for the prevention and/or therapeutic treatment of type 1 allergies. 
     
     
         20 . A pharmaceutical preparation comprising at least one hypoallergenic variant of  claim 9  and optionally further active compounds and/or assistants for the prevention and/or therapeutic treatment of type 1 allergies.