Cloned leptospira outer membrane protein

Abstract

An antigenic preparation is provided which contains a 63 Kd outer membrane protein from Leptospira which can be used immunologically as a vaccine for leptospirosis caused by this organism. Also provided in the invention are polynucleotides encoding the protein and antibodies which bind the protein which are useful in the diagnosis of leptospirosis.

1 . An isolated polypeptide comprising the amino acid sequence of OmpL2.  
     
     
         2 . The polypeptide of  claim 1 , wherein the polypeptide has a molecular weight of about 63 kD.  
     
     
         3 . The polypeptide of  claim 1 , wherein the polypeptide has essentially the amino acid sequence of FIG. 1.  
     
     
         4 . The polypeptide of  claim 1 , wherein the OmpL2 is from  Leptospira alstoni.    
     
     
         5 . The polypeptide of  claim 4 , wherein the OmpL2 is from a serovar of  Leptospira alstoni  selected the group consisting of  grippotyphosa  and Mozdok.  
     
     
         6 . The polypeptide of  claim 1 , wherein the OmpL2 is from  Leptospira interrogans.    
     
     
         7 . The polypeptide of  claim 6 , wherein the OmpL2 is from a serovar of  Leptospira interrogans  selected from the group consisting of  icterohaemorrhagiae, pomona  and  bratislava.    
     
     
         8 . An isolated polynucleotide sequence which encodes the polypeptide of  claim 1 .  
     
     
         9 . The polynucleotide sequence of  claim 8 , wherein the polynucleotide is DNA.  
     
     
         10 . The polynucleotide of  claim 8 , wherein the ompL2 sequence is selected from the group consisting of 
 a. the nucleotide sequence of FIG. 1, wherein T can also be U;    b. nucleic acid sequences complementary to the nucleotide sequence of FIG. 1; and    c. fragments of a. or b. that are at least 15 bases in length and which will selectively hybridize to genomic DNA which encodes the polypeptide of FIG. 1.    
     
     
         11 . The polynucleotide sequence of  claim 8 , wherein the polynucleotide is RNA.  
     
     
         12 . A recombinant expression vector containing the polynucleotide of  claim 8 .  
     
     
         13 . The expression vector of  claim 12 , wherein the vector is a plasmid.  
     
     
         14 . The vector of  claim 12 , wherein the polynucleotide sequence is from  L. alstoni.    
     
     
         15 . A host cell transformed with the expression vector of  claim 12 .  
     
     
         16 . The host cell of  claim 15 , wherein the cell is a prokaryote.  
     
     
         17 . The prokaryote of  claim 16 , which is  E. coli.    
     
     
         18 . The host cell of  claim 15 , wherein the cell is a eukaryote.  
     
     
         19 . A method of producing OmpL2 polypeptide which comprises: 
 a. transforming a host with the polynucleotide of claim  8 ; and    b. expressing the polynucleotide in the host.    
     
     
         20 . The method of  claim 19 , which further comprises isolating the OmpL2 polypeptide.  
     
     
         21 . The method of  claim 19 , wherein the host is a prokaryote.  
     
     
         22 . A pharmaceutical composition useful for inducing an immune response to pathogenic Leptospira in an animal comprising an immunogenically effective amount of OmpL2 in a pharmaceutically acceptable carrier.  
     
     
         23 . The pharmaceutical composition of  claim 22 , wherein the pharmaceutically acceptable carrier contains an adjuvant.  
     
     
         24 . A method of inducing an immune response to pathogenic Leptospira in an animal comprising immunizing the animal with the composition of  claim 22 .  
     
     
         25 . A pharmaceutical composition useful for inducing an immune response to pathogenic Leptospira in an animal comprising an immunogenically effective amount of antibody which binds OmpL2 in a pharmaceutically acceptable carrier.  
     
     
         26 . An antibody which binds to OmpL2.  
     
     
         27 . The antibody of  claim 26 , wherein the antibody is polyclonal.  
     
     
         28 . The antibody of  claim 26 , wherein the antibody is monoclonal.  
     
     
         29 . A method of detecting a pathogenic Leptospira in a sample comprising contacting a pathogen-specific cell component in the sample with a reagent which binds to the pathogen-specific cell component and detecting the binding of the reagent to the cell component.  
     
     
         30 . The method of  claim 29 , wherein the pathogen-specific cell component is nucleic acid which encodes OmpL2 polypeptide.  
     
     
         31 . The method of  claim 30 , wherein the nucleic acid is DNA.  
     
     
         32 . The method of  claim 29 , wherein the nucleic acid is RNA.  
     
     
         33 . The method of  claim 29 , wherein the pathogen specific cell component is OmpL2 polypeptide.  
     
     
         34 . The method of  claim 29 , wherein the reagent is a probe.  
     
     
         35 . The method of  claim 34 , wherein the probe is nucleic acid.  
     
     
         36 . The method of  claim 34 , wherein the probe is an antibody.  
     
     
         37 . The method of  claim 36 , wherein the antibody is polyclonal.  
     
     
         38 . The method of  claim 36 , wherein the antibody is monoclonal.  
     
     
         39 . The method of  claim 34 , wherein the probe is detectably labeled.  
     
     
         40 . The method of  claim 39 , wherein the label is selected from the group consisting of a radioisotope, a bioluminescent compound, a chemiluminescent compound, a fluorescent compound, a metal chelate, or an enzyme.  
     
     
         41 . The method of  claim 29 , wherein the sample is from an animal selected from the group consisting of human, swine and cattle.  
     
     
         42 . A method for detecting antibody to OmpL2 polypeptide in a sample which comprises contacting the sample with OmpL2 polypeptide under conditions which allow the antibody to bind to OmpL2 polypeptide and detecting the binding of the antibody to the OmpL2 polypeptide.  
     
     
         43 . The method of  claim 42 , wherein the OmpL2 polypeptide is detectably labelled.  
     
     
         44 . A kit useful for the detection of OmpL2 polypeptide, the kit comprising carrier means being compartmentalized to receive in close confinement therein one or more containers comprising a first container containing a OmpL2 binding reagent.  
     
     
         45 . The kit of  claim 44 , wherein the reagent is an antibody.  
     
     
         46 . The kit of  claim 45 , wherein the antibody is human.  
     
     
         47 . The kit of  claim 45 , wherein the antibody is monoclonal.  
     
     
         48 . A kit useful for the detection of OmpL2 polynucleotide, the kit comprising carrier means being compartmentalized to receive in close confinement therein one or more containers comprising a first container containing a OmpL2 polynucleotide binding reagent.  
     
     
         49 . The kit of  claim 48 , wherein the binding reagent is nucleic acid.  
     
     
         50 . A kit useful for the detection of antibody to OmpL2 polypeptide, the kit comprising carrier means being compartmentalized to receive in close confinement therein one or more containers comprising container containing OmpL2 polypeptide.