Helicobacter pylori proteins useful for vaccines and diagnostics

Abstract

This invention provides polypeptides of Helicobacter Pylori cytotoxin associated immunodominant (CAI) antigen. The invention also provides prophylactic and therapeutic vaccines comprising polypeptides of Helicobacter pylori CAI antigen, and methods for preparation. The invention also provides methods of treatment of Helicobacter pylori infection using these vaccines.

1 . An isolated polypeptide selected from the group consisting of: 
 (a) a  Helicobacter pylori  cytotoxin associated immunodominant antigen having the amino acid sequence shown in SEQ ID NO:5 modified by one or more conservative amino acid substitutions;    (b) a  Helicobacter pylori  cytotoxin associated immunodominant antigen comprising the amino acid sequence SEQ ID NO:25;    (c) a polypeptide that is immunologically identifiable by an antibody which reacts specifically with the  Helicobacter pylori  cytotoxin associated immunodominant antigen having the amino acid sequence of SEQ ID NO:5;    (d) a  Helicobacter pylori  cytotoxin associated immunodominant antigen comprising two repeats of the amino acid sequence of SEQ ID NO:12;    (e) a protein comprising a fragment of the amino acid sequence of SEQ ID NO:5, wherein said fragment comprises at least 11 amino acids of SEQ ID NO:5;    (f) the protein of (a), (b), (c), (d) or (e), comprising amino acid sequence SEQ ID NO:9;    (g) the protein of (a), (b), (c), (d) or (e), comprising amino acid sequence SEQ ID NO 10; and    (h) the protein of (a), (b), (c), (d) or (e), comprising amino acid sequence SEQ ID NO:23.    
     
     
         2 . The polypeptide of  claim 1 , which is conjugated to a detectable label  
     
     
         3 . The polypeptide of  claim 1 , which is coupled to a solid support.  
     
     
         4 . An immunogenic composition comprising the polypeptide of  claim 1  and a pharmaceutically acceptable carrier.  
     
     
         5 . The immunogenic composition of  claim 4 , wherein said carrier is an adjuvant.  
     
     
         6 . The immunogenic composition of  claim 4  wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:5.  
     
     
         7 . The immunogenic composition of  claim 4  wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:25.  
     
     
         8 . The immunogenic composition of  claim 4  wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:12.  
     
     
         9 . The immunogenic composition of  claim 4  wherein said polypeptide comprises at least 11 consecutive amino acids of SEQ ID NO:5 and wherein said polypeptide comprises at least one amino acid sequence selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:23.  
     
     
         10 . The immunogenic composition of  claim 4  further comprising at least one of: 
 (i) a  Helicobacter pylori  cytotoxic protein or precursor thereof;    (ii) a recombinant  Helicobacter pylori  heat shock protein;    (iii) a  Helicobacter pylori  urease.    
     
     
         11 . A therapeutic composition comprising the polypeptide of  claim 1  and a pharmaceutically acceptable carrier.  
     
     
         12 . The therapeutic composition of  claim 11  wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:5.  
     
     
         13 . The therapeutic composition of  claim 11  wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:25.  
     
     
         14 . The therapeutic composition of  claim 11  wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:12.  
     
     
         16 . The therapeutic composition of  claim 11  wherein said polypeptide comprises at least 11 consecutive amino acids of SEQ ID NO:5 and wherein said polypeptide comprises at least one amino acid sequence selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:23.  
     
     
         17 . The therapeutic composition of  claim 11  further comprising at least one of: 
 (i) a  Helicobacter pylori  cytotoxic protein or precursor thereof;    (ii) a recombinant  Helicobacter pylori  heat shock protein;    (iii) a  Helicobacter pylori  urease.    
     
     
         18 . A method for the preparation of an immunogenic composition according to  claim 4  comprising bringing one or more of said polypeptides into association with a pharmaceutically acceptable carrier and, optionally, an adjuvant.  
     
     
         19 . A method for the preparation of a therapeutic composition according to  claim 11  comprising bringing one or more of said polypeptides into association with a pharmaceutically acceptable carrier and, optionally, an adjuvant.  
     
     
         20 . A method for treating  Helicobacter pylori  infection comprising administering to a subject having a  Helicobacter pylori  infection a therapeutic amount of the composition of  claim 11 .  
     
     
         21 . The method of  claim 20  wherein said composition comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:5.  
     
     
         22 . The method of  claim 20  wherein said composition comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:25.  
     
     
         23 . The method of  claim 20  wherein said composition comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:12.  
     
     
         24 . The method of  claim 20  wherein said composition comprises a polypeptide comprising at least 11 consecutive amino acids of SEQ ID NO:5 and wherein said polypeptide comprises at least one amino acid sequence selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:23.  
     
     
         25 . The method of  claim 20  wherein said composition further comprising at least one of: 
 (i) a  Helicobacter pylori  cytotoxic protein or precursor thereof;    (ii) a recombinant  Helicobacter pylori  heat shock protein;    (iii) a  Helicobacter pylori  urease.    
     
     
         26 . A method of eliciting an immune response to a  Helicobacter pylori  protein comprising administering to a subject an immunogenic amount of a protein of  claim 1 .  
     
     
         27 . The method of  claim 26  wherein said protein comprises the amino acid sequence of SEQ ID NO:5.  
     
     
         28 . The method of  claim 26  wherein said protein comprises the amino acid sequence of SEQ ID NO:25.  
     
     
         29 . The method of  claim 26  wherein said protein comprises the amino acid sequence of SEQ ID NO:12.  
     
     
         30 . The method of  claim 26  wherein said protein comprises at least 11 contiguous amino acids of SEQ ID NO:5 and comprising at least one amino acid sequence selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:23.  
     
     
         31 . A method for diagnosing the presence of  Helicobacter pylori  in a sample comprising contacting a sample with an antibody that specifically binds to a protein of  claim 1  and detecting the binding of said antibody, wherein detection of specific binding of said antibody indicates the presence of  Helicobacter pylori  in said sample.  
     
     
         32 . The method of  claim 31  wherein said antibody specifically binds a protein having an amino acid sequence of SEQ ID NO:5.  
     
     
         33 . The method of  claim 31  wherein said antibody specifically binds a protein comprising the amino acid sequence of SEQ ID NO:25.  
     
     
         34 . The method of  claim 31  wherein said antibody specifically binds a protein comprising the amino acid sequence of SEQ ID NO:12.  
     
     
         35 . The method of  claim 31  wherein said antibody specifically binds a protein comprising at least 11 contiguous amino acids of SEQ ID NO:5 and having the at least one amino acid sequence selected from the group consisting of SEQ ID NO:9; SEQ ID NO:10; and SEQ ID NO:23.  
     
     
         36 . An in vitro immunodiagnostic assay comprising at least one step involving as at least one binding partner a protein according to  claim 1 .  
     
     
         37 . The immunodiagnostic assay of  claim 36  wherein said protein is conjugated to a label or coupled to a solid support.  
     
     
         38 . A kit for an immunodiagnostic assay, comprising at least one protein according to  claim 1 .  
     
     
         39 . A polynucleotide comprising a nucleotide sequence that encodes a protein according to  claim 1 .  
     
     
         40 . The polynucleotide of  claim 39 , comprising the nucleotide sequence of SEQ ID NO:5, or a fragment thereof.  
     
     
         41 . A polynucleotide probe comprising at least 8 contiguous nucleotides of (a) the polynucleotide of  claim 39  or  claim 40 , or (b) the complement of (a).  
     
     
         42 . The probe of  claim 41 , comprising at least 14 contiguous nucleotides of (a) or (b).  
     
     
         43 . A nucleic acid detection assay wherein at least one step comprises contacting a sample with the probe of  claim 39  or  40 .  
     
     
         44 . A polynucleotide amplification process for amplifying a  Helicobacter pylori  protein-encoding nucleic acid comprising contacting a sample with a plurality of polynucleotide primers wherein at least one primer is a polynucleotide comprising at least 14 contiguous nucleotides of (a) the polynucleotide of  claim 39  or  claim 40 , or (b) the complement of (a).