BASB029 polynucleotide(s) and polypeptides from Neisseria meningitidis

Abstract

The invention provides BASB029 polypeptides and polynucleotides encoding BASB029 polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are diagnostic, prophylactic and therapeutic uses.

What is claimed is:  
     
         1 . An isolated polypeptide comprising a member selected from the group consisting of 
 (a) an amino acid sequence which has at least 90% identity to SEQ ID NOs:2 or 4;    (b) an immunogenic fragment of the amino acid sequence of (a), wherein the immunogenic fragment is at least 90% identical to an aligned contiguous segment of SEQ ID NOs:2 or 4; and    (c) an immunogenic fragment of the amino acid sequence of (a) that matches an aligned contiguous segment of SEQ ID NOs:2 or 4 with no more than five single amino acid substitutions, deletions or additions;    wherein the isolated polypeptide, when administered to a subject in a suitable composition which can include an adjuvant, or a suitable carrier coupled to the polypeptide, induces an immune response that recognizes a polypeptide having the sequence of SEQ ID NOs:2 or 4.    
     
     
         2 . The isolated polypeptide of  claim 1 , wherein the immune response recognizes an antigen having molecular weight 65-70 kD (by SDS PAGE) in  Neisseria meningitidis  strains H44/76, M97 250987, BZ10, BZ198, EG328, and ATCC 13090.  
     
     
         3 . The isolated polypeptide of  claim 1 , wherein the immunogenic fragment of (b) comprises at least 15 amino acids.  
     
     
         4 . The isolated polypeptide of  claim 3 , wherein the immunogenic fragment matches an aligned contiguous segment of SEQ ID NOs:2 or 4 with no more than a single amino acid substitution, deletion or addition.  
     
     
         5 . The isolated polypeptide of  claim 1 , wherein the immunogenic fragment of (b) comprises at least 20 amino acids.  
     
     
         6 . The isolated polypeptide of  claim 1  wherein the amino acid sequence of (a) has at least 95% identity to SEQ ID NOs:2 or 4.  
     
     
         7 . The isolated polypeptide of  claim 6  wherein the isolated polypeptide comprises the amino acid sequence of SEQ ID NOs:2 or 4.  
     
     
         8 . The isolated polypeptide of  claim 6  wherein the isolated polypeptide consists of the amino acid sequence of SEQ ID NOs:2 or 4.  
     
     
         9 . A fusion protein comprising the isolated polypeptide of  claim 1 .  
     
     
         10 . The isolated polypeptide of  claim 1  wherein the polypeptide is the immunogenic fragment having no more than two single amino acid substitutions, deletions or additions relative to the aligned sequence.  
     
     
         11 . The isolated polypeptide of  claim 1  wherein the polypeptide is the immunogenic fragment having no more than one single amino acid substitution, deletion or addition relative to the aligned sequence.  
     
     
         12 . The isolated polypeptide of  claim 1  wherein the polypeptide is the immunogenic fragment which matches the aligned sequence.  
     
     
         13 . An isolated polypeptide encoded by an isolated first polynucleotide wherein the isolated first polynucleotide hybridizes under stringent conditions to a second polynucleotide which encodes the polypeptide of SEQ ID NOs:2 or 4; wherein stringent conditions comprise overnight incubation at 42° C. in a solution comprising: 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH7.6), 5× Denhardt's solution, 10% dextran sulfate, and 20 micrograms/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1×SSC at about 65° C.; wherein the isolated polypeptide, when administered to a subject in a suitable composition which can include an adjuvant, or a suitable carrier coupled to the polypeptide, induces an immune response that recognizes a polypeptide having the sequence of SEQ ID NOs:2 or 4.  
     
     
         14 . An isolated polynucleotide encoding a polypeptide of  claim 1  or the full complement to the isolated polynucleotide.  
     
     
         15 . An isolated polynucleotide encoding a polypeptide of  claim 1 , wherein the isolated polynucleotide encodes the polypeptide comprising SEQ ID NOs:2 or 4.  
     
     
         16 . An isolated polynucleotide comprising the polynucleotide of SEQ ID NOs:1 or 3.  
     
     
         17 . An isolated polynucleotide segment comprising a polynucleotide sequence or the full complement of the entire length of the polynucleotide sequence, wherein the polynucleotide sequence hybridizes to the full complement of SEQ ID NOs:1 or 3 minus the full complement of any terminal stop codon, wherein the hybridization conditions include incubation at 42° C. in a solution comprising: 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH7.6), 5× Denhardt's solution, 10% dextran sulfate, and 20 micrograms/ml denatured, sheared salmon sperm DNA, followed by washing in 0.1×SSC at 65° C.; and, wherein the polynucleotide sequence is identical to SEQ ID NOs:1 or 3 minus any terminal stop codon, except that, over the entire length corresponding to SEQ ID NO:1 or 3 minus any terminal stop codon, n n  nucleotides are substituted, inserted or deleted, wherein n n  satisfies the following expression  
         n   n   ≦x   n −( x   n   ·y )  
       wherein x n  is the total number of nucleotides in SEQ ID NOs:1 or 3 minus any terminal stop codon, y is at least 0.95, and wherein any non-integer product of x n  and y is rounded down to the nearest integer before subtracting the product from x n ; and wherein the polynucleotide sequence detects  Neisseria meningitidis.    
     
     
         18 . An expression vector comprising the isolated polynucleotide of  claim 14 .  
     
     
         19 . A host cell transformed with the expression vector of  claim 18 .  
     
     
         20 . A process of producing an isolated polypeptide comprising (a) culturing the host cell of  claim 19  under conditions sufficient for the production of the encoded polypeptide and (b) recovering the polypeptide.  
     
     
         21 . A nucleic acid vaccine comprising the isolated polynucleotide of  claim 14  and a pharmaceutically acceptable carrier.  
     
     
         22 . An isolated polynucleotide segment comprising a polynucleotide sequence or the full complement of the entire length of the polynucleotide sequence, wherein the polynucleotide sequence is identical to SEQ ID NOs:1 or 3 minus any terminal stop codon, except that, over the entire length corresponding to SEQ ID NOs:1 or 3 minus any terminal stop codon, n n  nucleotides are substituted, inserted or deleted, wherein n n  satisfies the following expression  
         n   n   ≦x   n −( x   n   ·y )  
       wherein x n  is the total number of nucleotides in SEQ ID NOs:1 or 3 minus any terminal stop codon, y is at least 0.90, and wherein any non-integer product of x n  and y is rounded down to the nearest integer before subtracting the product from x n ; and wherein the polynucleotide sequence detects  Neisseria meningitidis.    
     
     
         23 . The isolated polynucleotide of  claim 22  where y is at least 0.95.  
     
     
         24 . An expression vector comprising the isolated polynucleotide of  claim 22  which codes for a polypeptide that, when administered to a mammal which can include an adjuvant, or a suitable carrier coupled to the polypeptide, induces an immune response that recognizes a polypeptide having the sequence of SEQ ID NOs:2 or 4.  
     
     
         25 . A host cell transformed with the isolated polynucleotide or an expression vector comprising the isolated polynucleotide of  claim 22 .  
     
     
         26 . A process of producing an isolated polypeptide comprising (a) culturing the host cell of  claim 25  under conditions sufficient for the production of the encoded polypeptide and (b) recovering the polypeptide.  
     
     
         27 . A vaccine comprising the polypeptide of  claim 1  and a pharmaceutically acceptable carrier.  
     
     
         28 . The vaccine of  claim 27 , wherein the composition comprises at least one other  Neisseria meningitidis  antigen.  
     
     
         29 . An antibody immunospecific for the polypeptide or immunogenic fragment of  claim 1 .  
     
     
         30 . A method for inducing an immune response in a mammal comprising administration of the polypeptide of  claim 1 .  
     
     
         31 . A method of diagnosing a  Neisseria meningitidis  infection, comprising identifying a polypeptide of  claim 1 , or an antibody that is immunospecific for the polypeptide, present within a biological sample from an animal suspected of having such an infection.  
     
     
         32 . A method for inducing an immune response in a mammal comprising administration of the isolated polynucleotide of  claim 14 .  
     
     
         33 . A therapeutic composition useful in treating humans with  Neisseria meningitidis  comprising at least one antibody directed against the polypeptide of  claim 1  and a suitable pharmaceutical carrier.  
     
     
         34 . A process for expressing the polynucleotide of  claim 14  comprising transforming a host cell with the expression vector comprising the polynucleotide and culturing the host cell under conditions sufficient for expression of the polynucleotide.