Multiple variants of meningococcal protein nmb1870

Abstract

Meningococcal protein NMB 1870 has been described in the prior art. The inventors have found that NMB 1870 is an effective antigen for eliciting anti-meningococcal antibody responses, and that it is expressed across all meningococcal serogroups. Forty-two different NMB 1870 sequences have been identified, and these group into three variants. Serum raised against a given variant is bactericidal within the same variant group, but is not active against strains which express one of the other two variants i.e. there is intra-variant cross-protection, but not inter-variant cross-protection. For maximum cross-strain efficacy, therefore, the invention uses mixture comprising different variants of NMB 1870.

1 . A composition comprising at least two of the following antigens: (a) a first protein, comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 24 and/or comprising an amino acid sequence consisting of a fragment of at least 7 contiguous amino acids from SEQ ID NO: 24; (b) a second protein, comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 33 and/or comprising an amino acid sequence consisting of a fragment of at least 7 contiguous amino acids from SEQ ID NO: 33; and (c) a third protein, comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 41 and/or comprising an amino acid sequence consisting of a fragment of at least 7 contiguous amino acids from SEQ ID NO: 41.  
     
     
         2 . The composition of  claim 1 , wherein: protein (a) has less than 70% sequence identity to protein (b); protein (a) has less than 70% sequence identity to protein (c); and protein (b) has less than 70% sequence identity to protein (c).  
     
     
         3 . The composition of any preceding claim, wherein the composition can elicit a bactericidal response effective against each of serogroup B  N. meningitidis  strains MC58, 961-5945 and M1239.  
     
     
         4 . The composition of any preceding claim, wherein the composition can elicit an antibody response which is bactericidal against  N. meningitidis  strains in at least 2 of hypervirulent lineages ET-37, ET-5, cluster A4, lineage 3, subgroup I, subgroup III, and subgroup IV-1.  
     
     
         5 . The composition of any preceding claim, wherein one or more of the proteins is a lipoprotein.  
     
     
         6 . The composition of any preceding claim, wherein at least one of the proteins does not include the amino acid sequence TRSKP (SEQ ID NO: 70) or TRSKPV (SEQ ID NO: 71) within 10 amino acids of its N-terminus.  
     
     
         7 . The composition of any preceding claim, wherein at least one of the proteins does not include the amino acid sequence PSEPPFG (SEQ ID NO: 72) within 10 amino acids of its N-terminus.  
     
     
         8 . The composition of any preceding claim, wherein at least one of the proteins includes the amino acid sequence GGGG (SEQ ID NO: 73).  
     
     
         9 . The composition of any preceding claim, wherein at least one of the proteins is used in the form of a fusion protein.  
     
     
         10 . The composition of  claim 9 , wherein the fusion protein includes SEQ ID NO: 46 and/or the  H. influenzae  P4 lipoprotein leader sequence.  
     
     
         11 . The composition of any preceding claim, comprising one or more proteins comprising an amino acid sequence selected from SEQ ID NO s : 1-45, 77, 79-85, 87-94, and 123-142.  
     
     
         12 . A composition comprising a hybrid protein of formula NH 2 -A-[-X-L-] n -B—COOH, wherein: n is 2 or more; L is an optional linker amino acid sequence; A is an optional N-terminal amino acid sequence; B is an optional C-terminal amino acid sequence; and the X moieties include at least two of: (a) a first protein, comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 24 and/or comprising an amino acid sequence consisting of a fragment of at least 7 contiguous amino acids from SEQ ID NO: 24; (b) a second protein, comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 33 and/or comprising an amino acid sequence consisting of a fragment of at least 7 contiguous amino acids from SEQ ID NO: 33; and (c) a third protein, comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 41 and/or comprising an amino acid sequence consisting of a fragment of at least 7 contiguous amino acids from SEQ ID NO: 41.  
     
     
         13 . The composition of  claim 12 , wherein the hybrid protein comprises one of the following amino acid sequences: SEQ ID NO s : 79, 82, 83, 85, 87, 88, 89, 90 and 142.  
     
     
         14 . The composition of any preceding claim, including fewer than 15 antigens.  
     
     
         15 . The composition of any preceding claim, comprising a Neisserial antigen other than a NMB1870 protein.  
     
     
         16 . The composition of any preceding claim, comprising a vesicle prepared from  N. meningitidis.    
     
     
         17 . The composition of any preceding claim, comprising a saccharide antigen from  N. meningitidis  serogroup A, C, W135 and/or Y.  
     
     
         18 . The composition of  claim 17 , comprising a saccharide antigen from  N. meningitidis  serogroups A, C, W135 and Y.  
     
     
         19 . The composition of any preceding claim, comprising a saccharide antigen from  Haemophilus influenzae  type B.  
     
     
         20 . The composition of  claim 17 ,  claim 18  or  claim 19 , wherein the saccharide antigen(s) is/are conjugated to one or more carrier proteins.  
     
     
         21 . The composition of  claim 17 ,  claim 18 ,  claim 19  or  claim 20 , wherein the saccharide antigen(s) are oligosaccharides.  
     
     
         22 . The composition of any preceding claim, comprising an antigen from  Streptococcus pneumoniae.    
     
     
         23 . The composition of any one of  claims 17  to  22 , wherein the serogroup A saccharide antigen is modified saccharide in which one or more of the hydroxyl groups on the native saccharide has/have been replaced by a blocking group.  
     
     
         24 . The composition of any one of  claims 17  to  22 , wherein, where a serogroup A saccharide antigen contains n monosaccharide units, at least 50% of the monosaccharide units do not have —OH groups at both of positions 3 and 4.  
     
     
         25 . The composition of any one of  claims 17  to  22 , wherein the serogroup A saccharide comprises monosaccharide units, wherein at least 1 of the monosaccharide units does not have —OH at the 3 position and do not have —OH at the 4 position.  
     
     
         26 . The composition of any one of  claims 17  to  25 , wherein the serogroup A saccharides have the formula:  
       
         
           
           
               
               
           
         
       
       wherein 
 n is an integer from 1 to 100 (preferably an integer from 15 to 25);  
 T is of the formula (A) or (B):  
                     
 each Z group is independently selected from OH or a blocking group; and  
 each Q group is independently selected from OH or a blocking group;  
 Y is selected from OH or a blocking group;  
 E is H or a nitrogen protecting group;  
 and wherein more than 7% of the Q groups are blocking groups.  
 
     
     
         27 . The composition of  claim 20 , wherein the carrier protein is a diphtheria toxoid, a tetanus toxoid, CRM,97, a  N. meningitidis  outer membrane protein, protein D from  H. influenzae,  or pneumococcal surface protein PspA.  
     
     
         28 . The composition of any preceding claim, comprising: (i) at least two of said antigens (a), (b) and/or (c); (ii) a saccharide antigen from each of  N. meningitidis  serogroups A, C, W135 and Y; (iii) a saccharide antigen from  Haemophilus influenzae  type B; and (iv) an antigen from  Streptococcus pneumoniae.    
     
     
         29 . The composition of any preceding claim, for use as a medicament.  
     
     
         30 . A method for raising an antibody response in a mammal, comprising administering the composition of any preceding claim to the mammal.  
     
     
         31 . The method of  claim 30 , wherein the method protects a mammal against a Neisserial infection.  
     
     
         32 . The use of at least two of antigens (a), (b) and (c) as defined in  claim 1 , in the manufacture of a medicament for preventing Neisserial infection in a mammal.  
     
     
         33 . Nucleic acid encoding the protein of  claim 12  or  claim 13.