Modified surface antigen

Abstract

Novel proteins that constitute modified forms of a Neisseria meningitidis surface antigen and encoding nucleic acids are provided. The modified surface proteins are characterized by having deletions of non-conserved amino acids, and thereby being capable of eliciting cross-protective immune responses against Neisseria meningitidis . The invention extends to the use of the modified surface antigens in diagnostics, in therapeutic and prophylactic vaccines and in the design and/or screening of medicaments. The modified surface antigens are particularly useful in vaccines which effectively immunize against a broader spectrum of N. meningitidis strains than would be expected from a corresponding wild-type surface antigen.

We claim: 
     
         1 . A method of immunizing a mammal against  Neisseria meningitidis , comprising administering to the mammal an isolated protein comprising:
 (A) one or more conserved regions of an NhhA polypeptide, which one or more conserved regions respectively consist of any of the following amino acid sequences selected from the group consisting of:
 (i) residues 1 to 50 of SEQ ID NO:11; 
 (ii) residues 109 to120 of SEQ ID NO:11; 
 (iii) residues 135 to198 of SEQ ID NO:11; 
 (iv) residues 221 to 239 of SEQ ID NO:11; and 
 (v) residues 249 to 604 of SEQ ID NO:11; 
   and   (B) one or more variable regions of an NhhA polypeptide, the one or more variable regions respectively consisting of any of the following amino acid sequences selected from the group consisting of:
 (a) residues 121 to134 of SEQ ID NO:11; 
 (b) residues 199 to 220 of SEQ ID NO:11; and 
 (c) residues 240 to 248 of SEQ ID NO:11; 
   wherein a substantial portion variable region of residues 51-108 of SEQ ID NO:11 or a fragment consisting of at least six amino acids thereof is absent;   to thereby enable the mammal to elicit a protective immune response against  Neisseria meningitidis.      
     
     
         2 . A method of immunizing a mammal against  Neisseria meningitidis , comprising administering to the mammal an isolated protein consisting of two or more contiguous conserved regions of an NhhA polypeptide, the two or more conserved regions respectively consisting of any of the following amino acid sequences selected from the group consisting of:
 (i) residues 1 to 50 of SEQ ID NO:11;   (ii) residues 109 to 120 of SEQ ID NO:11;   (iii) residues 135 to198 of SEQ ID NO:11;   (iv) residues 221 to 239 of SEQ ID NO:11; and   (v) residues 249 to 604 of SEQ ID NO:11;   to thereby enable the mammal to elicit an immune response against  Neisseria meningitidis.      
     
     
         3 . The method of  claim 1 , wherein the protein is a deletion mutant of the NhhA polypeptide. 
     
     
         4 . The method of  claim 2 , wherein the protein is a deletion mutant of the NhhA polypeptide. 
     
     
         5 . The method of  claim 1 , wherein the conserved region amino acid sequence is selected from the group consisting of:
 (I) residues 1 to 50 of SEQ ID NO:1;   (II) residues 1 to 50 of SEQ ID NO:2;   (III) residues 1 to 50 of SEQ ID NO:3;   (IV) residues 1 to 50 of SEQ ID NO:4;   (V) residues 1 to 50 of SEQ ID NO:5;   (VI) residues 1 to 50 of SEQ ID NO:6;   (VII) residues 1 to 50 of SEQ ID NO:7;   (VIII) residues 1 to 50 of SEQ ID NO:8;   (IX) residues 1 to 50 of SEQ ID NO:9;   (X) residues 1 to 50 of SEQ ID NO:10;   (XI) residues 109 to 120 of SEQ ID NO:1;   (XII) residues 103 to 114 of SEQ ID NO:2;   (XIII) residues 106 to 117 of SEQ ID NO:3;   (XIV) residues 105 to 116 of SEQ ID NO:4;   (XV) residues 109 to 120 of SEQ ID NO:5;   (XVI) residues 106 to 117 of SEQ ID NO:6;   (XVII) residues 105 to 116 of SEQ ID NO:7;   (XVIII) residues 105 to 116 of SEQ ID NO:8;   (XIX) residues 105 to 116 of SEQ ID NO:9;   (XX) residues 103 to 114 of SEQ ID NO:10;   (XXI) residues 125 to 188 of SEQ ID NO:1;   (XXII) residues 125 to 188 of SEQ ID NO:2;   (XXIII) residues 122 to 185 of SEQ ID NO:3;   (XXIV) residues 127 to 190 of SEQ ID NO:4;   (XXV) residues 125 to 188 of SEQ ID NO:5;   (XXVI) residues 132 to 195 of SEQ ID NO:6;   (XXVII) residues 131 to 194 of SEQ ID NO:7;   (XXVIII) residues 131 to 194 of SEQ ID NO:8;   (XXIX) residues 127 to 190 of SEQ ID NO:9;   (XXX) residues 125 to 188 of SEQ ID NO:10;   (XXXI) residues 211 to 229 of SEQ ID NO:1;   (XXXII) residues 211 to 229 of SEQ ID NO:2;   (XXXIII) residues 206 to 224 of SEQ ID NO:3;   (XXXIV) residues 213 to 231 of SEQ ID NO:4;   (XXXV) residues 211 to 229 of SEQ ID NO:5;   (XXXVI) residues 218 to 236 of SEQ ID NO:6;   (XXXVII) residues 217 to 235 of SEQ ID NO:7;   (XXXVIII) residues 217 to 235 of SEQ ID NO:8;   (XXXIX) residues 213 to 231 of SEQ ID NO:9;   (XL) residues 209 to 227 of SEQ ID NO:10;   (XLI) residues 237 to 591 of SEQ ID NO:1;   (XLII) residues 237 to 592 of SEQ ID NO:2;   (XLIII) residues 235 to 589 of SEQ ID NO:3;   (XLIV) residues 239 to 594 of SEQ ID NO:4;   (XLV) residues 237 to 591 of SEQ ID NO:5;   (XLVI) residues 244 to 599 of SEQ ID NO:6;   (XLVII) residues 243 to 598 of SEQ ID NO:7;   (XLVIII) residues 243 to 598 of SEQ ID NO:8.   (XLIX) residues 239 to 594 of SEQ ID NO:9; and   (L) residues 237 to 592 of SEQ ID NO:10.   
     
     
         6 . The method of  claim 2 , wherein the conserved region amino acid sequence is selected from the group consisting of:
 (I) residues 1 to 50 of SEQ ID NO:1;   (II) residues 1 to 50 of SEQ ID NO:2;   (III) residues 1 to 50 of SEQ ID NO:3;   (IV) residues 1 to 50 of SEQ ID NO:4;   (V) residues 1 to 50 of SEQ ID NO:5;   (VI) residues 1 to 50 of SEQ ID NO:6;   (VII) residues 1 to 50 of SEQ ID NO:7;   (VIII) residues 1 to 50 of SEQ ID NO:8;   (IX) residues 1 to 50 of SEQ ID NO:9;   (X) residues 1 to 50 of SEQ ID NO:10;   (XI) residues 109 to 120 of SEQ ID NO:1;   (XII) residues 103 to 114 of SEQ ID NO:2;   (XIII) residues 106 to 117 of SEQ ID NO:3;   (XIV) residues 105 to 116 of SEQ ID NO:4;   (XV) residues 109 to 120 of SEQ ID NO:5;   (XVI) residues 106 to 117 of SEQ ID NO:6;   (XVII) residues 105 to 116 of SEQ ID NO:7;   (XVIII) residues 105 to 116 of SEQ ID NO:8;   (XIX) residues 105 to 116 of SEQ ID NO:9;   (XX) residues 103 to 114 of SEQ ID NO:10;   (XXI) residues 125 to 188 of SEQ ID NO:1;   (XXII) residues 125 to 188 of SEQ ID NO:2;   (XXIII) residues 122 to 185 of SEQ ID NO:3;   (XXIV) residues 127 to 190 of SEQ ID NO:4;   (XXV) residues 125 to 188 of SEQ ID NO:5;   (XXVI) residues 132 to 195 of SEQ ID NO:6;   (XXVII) residues 131 to 194 of SEQ ID NO:7;   (XXVIII) residues 131 to 194 of SEQ ID NO:8;   (XXIX) residues 127 to 190 of SEQ ID NO:9;   (XXX) residues 125 to 188 of SEQ ID NO:10;   (XXXI) residues 211 to 229 of SEQ ID NO:1;   (XXXII) residues 211 to 229 of SEQ ID NO:2;   (XXXIII) residues 206 to 224 of SEQ ID NO:3;   (XXXIV) residues 213 to 231 of SEQ ID NO:4;   (XXXV) residues 211 to 229 of SEQ ID NO:5;   (XXXVI) residues 218 to 236 of SEQ ID NO:6;   (XXXVII) residues 217 to 235 of SEQ ID NO:7;   (XXXVIII) residues 217 to 235 of SEQ ID NO:8;   (XXXIX) residues 213 to 231 of SEQ ID NO:9;   (XL) residues 209 to 227 of SEQ ID NO:10;   (XLI) residues 237 to 591 of SEQ ID NO:1;   (XLII) residues 237 to 592 of SEQ ID NO:2;   (XLIII) residues 235 to 589 of SEQ ID NO:3;   (XLIV) residues 239 to 594 of SEQ ID NO:4;   (XLV) residues 237 to 591 of SEQ ID NO:5;   (XLVI) residues 244 to 599 of SEQ ID NO:6;   (XLVII) residues 243 to 598 of SEQ ID NO:7;   (XLVIII) residues 243 to 598 of SEQ ID NO:8.   (XLIX) residues 239 to 594 of SEQ ID NO:9; and   (L) residues 237 to 592 of SEQ ID NO:10.   
     
     
         7 . The method of  claim 1 , wherein the variable region amino acid sequence is selected from the group consisting of:
 (AA) residues 121 to 124 of SEQ ID NO:1;   (AB) residues 115 to 124 of SEQ ID NO:2;   (AC) residues 118 to 121 of SEQ ID NO:3;   (AD) residues 117 to 126 of SEQ ID NO:4;   (AE) residues 121 to 124 of SEQ ID NO:5;   (AF) residues 118 to 131 of SEQ ID NO:6;   (AG) residues 117 to 130 of SEQ ID NO:7;   (AH) residues 117 to 130 of SEQ ID NO:8;   (AI) residues 117 to 126 of SEQ ID NO:9;   (AJ) residues 115 to 124 of SEQ ID NO:10;   (BA) residues 189 to 210 of SEQ ID NO:1;   (BB) residues 189 to 210 of SEQ ID NO:2;   (BC) residues 186 to 205 of SEQ ID NO:3;   (BD) residues 191 to 212 of SEQ ID NO:4;   (BE) residues 189 to 210 of SEQ ID NO:5;   (BF) residues 196 to 217 of SEQ ID NO:6;   (BG) residues 195 to 216 of SEQ ID NO:7;   (BH) residues 195 to 216 of SEQ ID NO:8;   (BI) residues 191 to 212 of SEQ ID NO:9;   (BJ) residues 189 to 208 of SEQ ID NO:10;   (CA) residues 230 to 236 of SEQ ID NO:1;   (CB) residues 230 to 236 of SEQ ID NO:2;   (CC) residues 225 to 234 of SEQ ID NO:3;   (CD) residues 232 to 238 of SEQ ID NO:4;   (CE) residues 230 to 236 of SEQ ID NO:5;   (CF) residues 237 to 243 of SEQ ID NO:6;   (CG) residues 236 to 242 of SEQ ID NO:7;   (CH) residues 236 to 242 of SEQ ID NO:8;   (CI) residues 232 to 238 of SEQ ID NO:9; and   (CJ) residues 228 to 236 of SEQ ID NO:10.   
     
     
         8 . The method of  claim 1 , wherein the isolated protein comprises an amino acid sequence selected from the group consisting of: SEQ ID NO:23; SEQ ID NO:24; SEQ ID NO:26; SEQ ID NO:33; SEQ ID NO:36 and SEQ ID NO:38. 
     
     
         9 . The method of  claim 2 , comprising an amino acid sequence selected from the group consisting of SEQ ID NO:25; SEQ ID NO:27; SEQ ID NO:37 and SEQ ID NO:39. 
     
     
         10 . The method of  claim 1 , wherein the isolated protein comprises a fusion partner. 
     
     
         11 . The method of  claim 2 , wherein the isolated protein comprises a fusion partner. 
     
     
         12 . The method of  claim 1 , wherein the isolated protein is administered in a pharmaceutical composition that comprises a pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         13 . The method of  claim 2 , wherein the isolated protein is administered in a pharmaceutical composition that comprises a pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         14 . The method of  claim 12 , wherein the pharmaceutical composition comprises an adjuvant. 
     
     
         15 . The method of  claim 13 , wherein the pharmaceutical composition comprises an adjuvant. 
     
     
         16 . The method of  claim 1 , which enables the mammal to elict a protective immune response to a plurality of strains of  N. meningitidis.    
     
     
         17 . The method of  claim 2 , which enables the mammal to elict a protective immune response to a plurality of strains of  N. meningitidis.    
     
     
         18 . The method of  claim 1 , wherein the mammal is a human. 
     
     
         19 . The method of  claim 2 , wherein the mammal is a human.