ME-5, ME-2, and EPP2: human protein antigens reactive with autoantibodies present in the serum of women suffering from endometriosis

Abstract

Three human endometrial antigens, ME-5, ME-2 and EPP2, and combination thereof have been discovered that are the targets of antibodies present in the serum of women suffering from endometriosis. These protein products and the human antibodies reactive with them are used as tools for diagnostic assays that monitor patients with the disease of endometriosis.

1 . A recombinant polynucleotide comprising an isolated nucleotide sequence from SEQ ID NO:2 encoding a polypeptide epitope of at least 5 amino acids of ME-5 (SEQ ID NO:3), wherein the epitope specifically binds to antibodies from subjects diagnosed with endometriosis.  
     
     
         2 . The polynucleotide of  claim 1  wherein the nucleotide sequence is selected from the ME-5 sequence of SEQ ID NO:1.  
     
     
         3 . The polynucleotide of  claim 1  wherein the nucleotide sequence is a native ME-5 nucleotide sequence.  
     
     
         4 . The polynucleotide of  claim 1  wherein the nucleotide sequence is substantially identical to nucleotides 1 to 912 of SEQ ID NO:2.  
     
     
         5 . A purified, recombinant ME-5 polypeptide whose amino acid sequence is substantially identical to that of SEQ ID NO:3 or an allelic variant of SEQ ID NO:3.  
     
     
         6 . A purified polypeptide comprising an epitope of at least 5 amino acids of ME-5 (SEQ ID NO:3), wherein the epitope specifically binds to antibodies from subjects diagnosed with endometriosis.  
     
     
         7 . A composition consisting essentially of an antibody that specifically binds to an epitope of ME-5 polypeptide (SEQ ID NO:3).  
     
     
         8 . The composition of  claim 7  wherein the antibody is a monoclonal antibody.  
     
     
         9 . A method for detecting a ME-5 polypeptide (SEQ ID NO:3) in a sample, comprising the steps of: 
 (1) contacting the sample with an antibody that specifically binds to an epitope of the ME-5 polypeptide and    (2) detecting specific binding between the antibody and ME-5 polypeptide;    whereby specific binding provides a detection of ME-5 polypeptide in the sample.    
     
     
         10 . A method for diagnosing endometriosis in a human subject comprising the steps of: 
 (a) detecting a test amount of an antibody that specifically binds to ME-5 (SEQ ID NO:3) polypeptide in a sample from the subject; and    (b) comparing the test amount with a normal range of the antibody in a control sample from a subject who does not suffer from endometriosis,    whereby a test amount above the normal range provides a positive indication in the diagnosis of endometriosis.    
     
     
         11 . The method of  claim 10  wherein the sample comprises blood serum.  
     
     
         12 . The method  claim 10  wherein the step of detecting comprises capturing the antibody from the sample with an immobilized ME-5 or a peptide comprising an epitope of ME-5 and detecting the captured antibody.  
     
     
         13 . A recombinant polynucleotide comprising an isolated nucleotide sequence from SEQ ID NO:5 encoding a polypeptide epitope of at least 5 amino acids of ME-2 (SEQ ID NO:6), wherein the epitope specifically binds to antibodies from subjects diagnosed with endometriosis.  
     
     
         14 . The polynucleotide of  claim 13  wherein the nucleotide sequence is selected from the ME-2 sequence of SEQ ID NO:4.  
     
     
         15 . The polynucleotide of  claim 13  wherein the nucleotide sequence is a native ME-2 nucleotide sequence.  
     
     
         16 . The polynucleotide of  claim 13  wherein the nucleotide sequence is substantially identical to nucleotides 1 to 1182 of SEQ ID NO:5.  
     
     
         17 . A purified, recombinant ME-2 polypeptide whose amino acid sequence is substantially identical to that of SEQ ID NO:6 or an allelic variant of SEQ ID NO:6.  
     
     
         18 . A purified polypeptide comprising an epitope of at least 5 amino acids of ME-2 (SEQ ID NO:6), wherein the epitope specifically binds to antibodies from subjects diagnosed with endometriosis.  
     
     
         19 . A composition consisting essentially of an antibody that specifically binds to ME-2 polypeptide (SEQ ID NO:6).  
     
     
         20 . The composition of  claim 19  wherein the antibodies are monoclonal antibodies.  
     
     
         21 . A method for detecting a ME-2 polypeptide (SEQ ID NO:6) in a sample, comprising the steps of: 
 (a) contacting the sample with an antibody that specifically binds to the ME-2 polypeptide and    (b) detecting specific binding between the antibody and ME-2 polypeptide;    whereby specific binding provides a detection of ME-2 polypeptide in the sample.    
     
     
         22 . A method for diagnosing endometriosis in a human subject comprising the steps of: 
 (a) detecting a test amount of an antibody that specifically binds to ME-2 polypeptide (SEQ ID NO:6) in a sample from the subject; and    (b) comparing the test amount with a normal range of the antibody in a control sample from a subject who does not suffer from endometriosis,    whereby a test amount above the normal range provides a positive indication in the diagnosis of endometriosis.    
     
     
         23 . The method of  claim 22  wherein the sample comprises blood serum.  
     
     
         24 . The method of  claim 22  wherein the step of detecting comprises capturing the antibody from the sample with an immobilized ME-2 or a peptide comprising an epitope of ME-2 and detecting captured antibody.  
     
     
         25 . A recombinant polynucleotide comprising an isolated nucleotide sequence from SEQ ID NO:8 encoding a polypeptide epitope of at least 5 amino acids of EPP2 (SEQ ID NO:9), wherein the epitope specifically binds to antibodies from subjects diagnosed with endometriosis.  
     
     
         26 . The polynucleotide of  claim 25  wherein the nucleotide sequence is selected from the EPP2 sequence of SEQ ID NO:7.  
     
     
         27 . The polynucleotide of  claim 25  wherein the nucleotide sequence is a native EPP2 nucleotide sequence.  
     
     
         28 . The polynucleotide of  claim 25  wherein the nucleotide sequence is substantially identical to nucleotides 1 to 300 of SEQ ID NO:8.  
     
     
         29 . A purified, recombinant EPP2 polypeptide whose amino acid sequence is substantially identical to that of SEQ ID NO:9 or an allelic variant of SEQ ID NO:9.  
     
     
         30 . A purified polypeptide comprising an epitope of at least 5 amino acids of EPP2 (SEQ ID NO:9), wherein the epitope specifically binds to antibodies from subjects diagnosed with endometriosis.  
     
     
         31 . A composition consisting essentially of an antibody that specifically binds to EPP2 polypeptide (SEQ ID NO:9).  
     
     
         32 . The composition of  claim 31  wherein the antibody is a monoclonal antibody.  
     
     
         33 . A method for detecting a EPP2 polypeptide (SEQ ID NO:9) in a human sample, comprising the steps of: 
 (a) contacting the sample with an antibody that specifically binds to an epitope of the EPP2 polypeptide and    (b) detecting specific binding between the antibody and EPP2 polypeptide;    whereby specific binding provides a detection of EPP2 polypeptide in the sample.    
     
     
         34 . A method for diagnosing endometriosis in a subject comprising the steps of: 
 (a) detecting a test amount of an antibody that specifically binds to EPP2 (SEQ ID NO:9) polypeptide or a peptide comprising an epitope of EPP2 in a sample from the subject; and    (b) comparing the test amount with a normal range of the antibody in a control sample from a subject who does not suffer from endometriosis,    whereby a test amount above the normal range provides a positive indication in the diagnosis of endometriosis.    
     
     
         35 . The method of  claim 34  wherein the sample comprises blood serum.  
     
     
         36 . The method of  claim 34  wherein the step of detecting comprises capturing the antibody from the sample with an immobilized EPP2 or a peptide comprising an epitope of EPP2 and detecting captured antibody.  
     
     
         37 . A composition containing at least one of 
 a purified, recombinant ME-5 polypeptide whose amino acid sequence is substantially identical to that of SEQ ID NO:3 or an allelic variant of SEQ ID NO:3;    a purified, recombinant ME-2 polypeptide whose amino acid sequence is substantially identical to that of SEQ ID NO:6 or an allelic variant of SEQ ID NO:6; and    a purified, recombinant EPP2 polypeptide whose amino acid sequence is substantially identical to that of SEQ ID NO:9 or an allelic variant of SEQ ID NO:9.    
     
     
         38 . A composition containing at least one of 
 a purified polypeptide comprising an epitope of at least 5 amino acids of ME-5 (SEQ ID NO:3);    a purified polypeptide comprising an epitope of at least 5 amino acids of ME-2 (SEQ ID NO:6), and    a purified polypeptide comprising an epitope of at least 5 amino acids of EPP2 (SEQ ID NO:9),    wherein said epitopes specifically bind to antibodies from subjects diagnosed with endometriosis.    
     
     
         39 . The compositions of claims  37  and  38  containing all three said polypeptides.  
     
     
         40 . A method for diagnosing endometriosis in a human subject comprising the steps of: 
 (a) detecting a test amount of an antibody that specifically binds to at least one of ME-5 (SEQ ID NO:3) polypeptide, ME-2 (SEQ ID NO:6) polypeptide, and EPP2 (SEQ ID NO:9) polypeptide in a sample from the subject; and    (b) comparing the test amount with a normal range of the antibody in a control sample from a subject who does not suffer from endometriosis,    whereby a test amount above the normal range provides a positive indication in the diagnosis of endometriosis.    
     
     
         41 . The method of  claim 40  wherein the sample comprises blood serum.  
     
     
         42 . The method of  claim 40  wherein the step of detecting comprises capturing the antibody from the sample with an immobilized peptide where said peptide is selected from the group consisting of ME-2 or a peptide comprising an epitope of ME-2, ME-5 or a peptide comprising an epitope of ME-5 and EPP2 or a peptide comprising an epitope of EPP2 and detecting captured antibody.  
     
     
         43 . The method of  claim 39  wherein binding polypeptide includes ME-2, ME-5 and EPP2.