Proteins with repetitive bacterial-Ig-like (big) domains present in leptospira species

Abstract

The invention relates to three isolated DNA molecules that encode for proteins, BigL1, BigL2 and BigL3, in the Leptospira sp bacterium which have repetitive Bacterial-Ig-like (Big) domains and their use in diagnostic, therapeutic and vaccine applications. According to the present invention, the isolated molecules encoding for BigL1, BigL2 and BigL3 proteins are used for the diagnosis and prevention of infection with Leptospira species that are capable of producing disease in humans and other mammals, including those of veterinary importance.

We claim:  
     
         1 . A substantially purified polypeptide having an amino acid sequence as set forth in SEQ ID NO:2 or functionally equivalent sequences thereof.  
     
     
         2 . An isolated polynucleotide segment encoding an amino acid sequence as set forth in SEQ ID NO:2 or functionally equivalent sequences thereof.  
     
     
         3 . An isolated polynucleotide selected from SEQ ID NO:1 or functionally equivalent sequences thereof.  
     
     
         4 . An substantially purified polypeptide having a amino acid sequence as set forth in SEQ ID NO:4 or functionally equivalent sequences thereof.  
     
     
         5 . An isolated polynucleotide encoding an amino acid sequence as set forth in SEQ ID NO:4 or functionally equivalent sequences thereof.  
     
     
         6 . An isolated polynucleotide selected from SEQ ID NO:3 or functionally equivalent sequences thereof.  
     
     
         7 . A substantially purified polypeptide having a amino acid sequence as set forth in SEQ ID NO:6 or functionally equivalent sequence thereof.  
     
     
         8 . An isolated polynucleotide encoding an amino acid sequence as set forth in SEQ ID NO:6 or functionally equivalent sequences thereof.  
     
     
         9 . An isolated polynucleotide selected from SEQ ID NO:5 or functionally equivalent sequences thereof.  
     
     
         10 . The polynucleotide of claims  2 ,  5  or  8  wherein the polynucleotide sequence is from Leptospira species.  
     
     
         11 . An substantially purified polypeptide sequence selected from SEQ ID NO: 8 or functionally equivalent sequences thereof.  
     
     
         12 . An isolated polynucleotide segment encoding an amino acid sequence as set forth in SEQ ID NO: 8 or functionally equivalent sequences thereof.  
     
     
         13 . An isolated polynucleotide selected from SEQ ID NO: 7 or functionally equivalent sequences thereof.  
     
     
         14 . An substantially purified polypeptide sequence selected from SEQ ID NO: 10 or functionally equivalent sequences thereof.  
     
     
         15 . An isolated polynucleotide segment encoding an amino acid sequence as set forth in SEQ ID NO: 10 or functionally equivalent sequence thereof.  
     
     
         16 . An isolated polynucleotide selected from SEQ ID NO: 9 or functionally equivalent sequences thereof.  
     
     
         17 . Antibodies that binds to the substantially purified polypeptide in claims  1 ,  4 ,  7 ,  11  and  14 , or functionally equivalent sequences.  
     
     
         18 . The pharmaceutical composition used to induce an immune response to pathogenic spirochete in a mammalian subject comprising of an immunogenically effective amount of the substantially purified polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof in a pharmaceutically acceptable carrier.  
     
     
         19 . The pharmaceutical composition of  claim 18  wherein the pharmaceutically acceptable carrier contains an adjuvant.  
     
     
         20 . The pharmaceutical composition of  claim 18  wherein the pathogenic spirochete is selected from Treponema, Borrelia and Leptospira.  
     
     
         21 . The pharmaceutical composition useful for providing an immune response to pathogenic spirochetes in a mammalian subject comprising an immunologically effective amount of antibody that binds polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof, in a pharmaceutically acceptable carrier.  
     
     
         22 . The method of inducing an immune response against infection with a pathogenic Leptospira in mammalian subjects comprising of administering to a pharmaceutical composition of  claim 18 .  
     
     
         23 . The method of inducing an immune response to a pathogenic spirochete in mammalian subject comprising administering to the individual a pharmaceutical composition of  claim 19 .  
     
     
         24 . A method for detecting pathogens in a sample which comprises contacting a sample suspected of containing a pathogenic spirochete with a reagent that binds to the pathogen-specific cell component and detecting binding of the reagent to the component.  
     
     
         25 . The method according to  claim 24  wherein the reagent that binds to the pathogen-specific cell component is an oligonucleotide for the identification of bigL1, bigL2 and bigL3 polynucleotide.  
     
     
         26 . The method according to  claim 24  wherein the reagent that binds to the pathogen-specific cell component is an antibody against the BigL1, BigL2 or BigL3 polypeptide or polypeptides with functionally equivalent sequences.  
     
     
         27 . The method of  claim 22  wherein the spirochete-specific component is a polynucleotide that encodes for polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof.  
     
     
         28 . The method of  claim 22  wherein the spirochete-specific component is polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof.  
     
     
         29 . The method of detecting antibodies to polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof in a sample, which comprises of contacting the sample with polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof under conditions which allow the antibody to bind with polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof, and detecting the binding of the antibody to polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof.  
     
     
         30 . A method of detecting polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof in a sample, comprising of contacting the sample with antibodies to polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequences thereof, which allow the polypeptides in the sample to bind to the antibodies and measuring the binding of the polypeptides to the said antibodies.  
     
     
         31 . A method of detecting nucleic acid encoding BigL1, BigL2, BigL3 or functionally equivalent sequences thereof in a sample, which uses polynucleotides that encodes for polypeptides in claims  1 ,  4 ,  7 ,  11  and  14  or functionally equivalent sequence thereof.  
     
     
         32 . A kit useful for the detection of BigL1, BigL2, and BigL3 polypeptides or polypeptides with functionally equivalent sequences; bigL1, bigL2 and bigL3 polynucleotides; or antibodies that bind to BigL1, BigL2, BigL3, polypeptides or polypeptides with functionally equivalent sequences.  
     
     
         33 . A kit useful for the detection of nucleic acid encoding BigL1, BigL2, BigL3 or functionally equivalent sequence thereof, the kit comprising carrier means containing one or more containers comprising a first container containing a polynucleotide that hybridizes to BigL1, BigL2, BigL3 nucleic acid or functionally equivalent sequence thereof.  
     
     
         34 . A kit useful for the detection of antibody to BigL1, BigL2, BigL3 nucleic acid or functionally equivalent sequence thereof, the kit comprising carrier means containing one or more containers comprising a first container containing BigL1, BigL2 or BigL3 polypeptide or functionally equivalent sequence thereof.