Haemophilus Influenzae Type IV PILI

Abstract

The invention described herein relates to a Haemophilus influenzae ( H. influenzae ) regulon encoding type IV pili. In particular, the invention relates to type IV pili from nontypeable H. influenzae (NTHi) and from H. influenzae strains a, b, c, e and f. The invention provides isolated H. influenzae pilus polynucleotides and polypeptides encoded by the polynucleotides as well as polynucleotides and polypeptides encoded by the polynucleotides involved in the assembly/disassembly of the structure. The invention also relates to uses of these polynucleotides and/or polypeptides including methods for eliciting an immune response to H. influenzae and methods of treating and preventing H. influenzae related pathological conditions.

1 . An isolated polynucleotide comprising a nucleotide sequence encoding an amino acid sequence of any one of: PilA polypeptide SEQ ID NO: 2, PilB polypeptide SEQ ID NO: 4, PilC polypeptide SEQ ID NO: 6, PilD polypeptide SEQ ID NO: 8, ComA polypeptide SEQ ID NO: 10, ComB polypeptide SEQ ID NO: 12, ComC polypeptide SEQ ID NO: 14, ComD polypeptide SEQ ID NO: 16, ComE polypeptide SEQ ID NO: 18, ComF polypeptide SEQ ID NO: 20, PilF polypeptide SEQ ID NO: 22, PilA polypeptide SEQ ID NO: 26, PilA polypeptide SEQ ID NO: 26, PilA polypeptide SEQ ID NO: 28, PilA polypeptide SEQ ID NO: 30, PilA polypeptide SEQ ID NO: 32, PilA polypeptide SEQ ID NO: 34, PilA polypeptide SEQ ID NO: 36, PilA polypeptide SEQ ID NO: 38, PilA polypeptide SEQ ID NO: 40, PilA polypeptide SEQ ID NO: 42, or pilA polypeptide SEQ ID NO: 44. 
     
     
         2 . An isolated polynucleotide comprising a nucleotide sequence of any one of: pilA SEQ ID NO: 1, pilB SEQ ID NO: 3, pilC SEQ ID NO: 5, pilD SEQ ID NO: 7, comA SEQ ID NO: 9, comB SEQ ID NO: 11, comC SEQ ID NO: 13, comD SEQ ID NO: 15, comE SEQ ID NO: 17, comF SEQ ID NO: 19, pilF SEQ ID NO: 21, pilA SEQ ID No: 25, pilA SEQ ID No: 27, pilA SEQ ID No: 29, pilA SEQ ID No: 31, pilA SEQ ID No: 33, pilA SEQ ID No: 35, pilA SEQ ID No: 37, pilA SEQ ID No: 39, pilA SEQ ID No: 41 or pilA SEQ ID No: 43. 
     
     
         3 . A vector comprising a polynucleotide of  claim 1 . 
     
     
         4 . An isolated polypeptide comprising an amino acid sequence encoded by a nucleotide sequence of  claim 1  or a fragment thereof. 
     
     
         5 . An isolated polypeptide comprising an amino acid sequence of any one of PilA polypeptide SEQ ID NO: 2, PilB polypeptide SEQ ID NO: 4, PilC polypeptide SEQ ID NO: 6, PilD polypeptide SEQ ID NO: 8, ComA polypeptide SEQ ID NO: 10, ComB polypeptide SEQ ID NO: 12, ComC polypeptide SEQ ID NO: 14, ComD polypeptide SEQ ID NO: 16, ComE polypeptide SEQ ID NO: 18, ComF polypeptide SEQ ID NO: 20, PilF polypeptide SEQ ID NO: 22, PilA polypeptide SEQ ID NO: 26, PilA polypeptide SEQ ID NO: 26, PilA polypeptide SEQ ID NO: 28, PilA polypeptide SEQ ID NO: 30, PilA polypeptide SEQ ID NO: 32, PilA polypeptide SEQ ID NO: 34, PilA polypeptide SEQ ID NO: 36, PilA polypeptide SEQ ID NO: 38, PilA polypeptide SEQ ID NO: 40, PilA polypeptide SEQ ID NO: 42, or pilA polypeptide SEQ ID NO: 44, or a peptide fragment thereof. 
     
     
         6 . A composition comprising a polypeptide or peptide fragment of  claim 5  and a pharmaceutically acceptable carrier. 
     
     
         7 . An antibody that specifically binds to a polypeptide or peptide fragment of  claim 5 . 
     
     
         8 . A composition comprising an antibody of  claim 8  and a pharmaceutically acceptable carrier. 
     
     
         9 . A method for detecting NTHi bacteria in a biological sample comprising
 (a) contacting a polynucleotide of  claim 1  or a fragment thereof with a biological sample, and   (b) detecting hybridization of the polynucleotide within the sample, wherein hybridization indicates the presence of NTHi bacteria.   
     
     
         10 . A method for detecting NTHi bacteria in a biological sample comprising
 (a) contacting a polynucleotide of  claim 2  or a fragment thereof with a biological sample, and   (b) detecting hybridization of the polynucleotide within the sample, wherein hybridization indicates the presence of NTHi bacteria.   
     
     
         11 . A method for detecting NTHi bacteria in a biological sample comprising:
 (a) contacting an antibody of  claim 7  with a biological sample, and   (b) detecting binding of the antibody within the sample, wherein binding indicates the presence of NTHi bacteria.   
     
     
         12 . The method of  claim 9  wherein the biological sample is selected from the group consisting of serum, sputum, ear fluid, blood, urine, lymphatic fluid, and cerebrospinal fluid. 
     
     
         13 . A method for eliciting an immune response to NTHi bacteria comprising administering an immunogenic dose of one or more polypeptides or peptide fragments of  claim 5  to a patient at risk of NTHi bacterial infection. 
     
     
         14 . A method for eliciting an immune response to NTHi bacteria comprising administering an immunogenic dose of one or more polynucleotides of  claim 1  to a patient at risk of NTHi bacterial infection. 
     
     
         15 . The method of  claim 14  wherein the polynucleotide is in a plasmid or viral vector. 
     
     
         16 . A method of treating or preventing NTHi bacterial infection comprising administering a molecule that inhibits expression or activity of a polypeptide of  claim 5  to an patient in need thereof. 
     
     
         17 . The method of  claim 16  wherein the molecule administered to the patient in need is an antisense oligonucleotide. 
     
     
         18 . The method of  claim 16  wherein the molecule administered to the patient in need is an antibody. 
     
     
         19 . The method of  claim 16  wherein the molecule administered to the patient in need is a small molecule. 
     
     
         20 . The method of  claim 16  wherein the NTHi infection is in the middle ear.