Anti-pdef antibodies and uses thereof

Abstract

The present invention relates to an isolated antibody or antigen-binding fragment thereof that specifically binds with high affinity to at least a portion of a segment of human prostate-derived Ets transcription factor (PDEF). The anti-PDEF antibody of the present invention is effective in prognostic and diagnostic assays for detecting PDEF with immunohistochemistry. The present invention also relates to methods of making the anti-PDEF antibody disclosed herein. The present invention further relates to vaccines against cancers associated with positive expression of PDEF, as well as methods for treating those cancers. Vectors, diagnostic kits, and hybridomas are also disclosed.

1 . An isolated antibody or antigen-binding fragment thereof that specifically binds with high affinity to at least a portion of a segment of a human prostate-derived Ets transcription factor (PDEF), wherein said segment comprises:
 (a) amino acid residues 1-104 of SEQ ID NO:1;   (b) amino acid residues 105-141 of SEQ ID NO:1; or   (c) amino acid residues 214-247 of SEQ ID NO:1.   
     
     
         2 . The antibody or antigen-binding fragment thereof according to  claim 1 , wherein said antigen-binding fragment is selected from the group consisting of an F(ab) 2  fragment, an Fab′ fragment, an Fab fragment, and an Fv fragment. 
     
     
         3 . The antibody or antigen-binding fragment thereof according to  claim 1 , wherein said antibody or antigen-binding fragment thereof is selected from the group consisting of a polyclonal antibody, a monoclonal antibody, and a single chain antibody. 
     
     
         4 . The antibody or antigen-binding fragment thereof of  claim 1 , wherein said antibody or antigen-binding fragment thereof is detectably labeled. 
     
     
         5 . The antibody or antigen-binding fragment thereof of  claim 4 , wherein the detectable label is selected from the group consisting of a radioisotope, an affinity label, an enzymatic label, a fluorescent label, and a paramagnetic atom. 
     
     
         6 . The antibody or antigen-binding fragment thereof according to  claim 1 , wherein the antibody or antigen-binding fragment thereof exhibits:
 (a) strong staining intensity as determined in an immunohistochemical assay with three or less of the normal tissues being selected from the group consisting of prostate gland tissue, salivary gland tissue, and trachea/bronchus tissue; and   (b) negative staining intensity as determined by an immunohistochemical assay with three or more tissues selected from the group consisting of adrenal gland tissue, blood, brain tissue, bone marrow, heart tissue, liver tissue, lung tissue, lymph node tissue, kidney tissue, ovary tissue, placenta tissue, spleen tissue, skeletal muscle tissue, thymus tissue, and testis tissue.   
     
     
         7 . A hybridoma that produces the antibody according to  claim 1 , wherein said antibody is a monoclonal antibody. 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . A vaccine for immunizing an individual against a cancer disease associated with positive expression of PDEF, said vaccine comprising:
 (a) a polypeptide comprising a PDEF fragment that is effective to induce an immune response to PDEF in the individual, wherein said PDEF fragment comprises:
 (i) an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4; 
 (ii) an amino acid sequence having at least 8 continuous residues from SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; or 
 (iii) two or more amino acid sequences having at least 8 continuous residues from SEQ ID NO:2, SEQ ID NO:3, and/or SEQ ID NO:4; and 
   (b) a pharmacologically acceptable carrier or adjuvant.   
     
     
         11 . The vaccine according to  claim 10  further comprising a nanoparticle vaccine comprising a peptide of at least 8 continuous residues from SEQ ID NO:2, SEQ ID NO:3, and/or SEQ ID NO:4. 
     
     
         12 . The vaccine according to  claim 10 , wherein said cancer disease is selected from the group consisting of breast cancer, prostate cancer, ovarian cancer, endometrial cancer, colon cancer, and cervical cancer. 
     
     
         13 . A method for immunizing an individual against a cancer disease associated with positive PDEF expression, said method comprising:
 administering the vaccine according to  claim 10  into the individual, wherein said individual is at high risk for, predisposed to, susceptible to, and/or diagnosed with said cancer disease associated with positive PDEF expression.   
     
     
         14 . The method according to  claim 13 , wherein said cancer disease is selected from the group consisting of breast cancer, prostate cancer, ovarian cancer, endometrial cancer, colon cancer, and cervical cancer. 
     
     
         15 . A viral or plasmid expression vector comprising:
 a nucleotide sequence that is operably linked to a promoter and that encodes an antigen, wherein said antigen is a PDEF fragment comprising:
 (i) an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4; 
 (ii) an amino acid sequence having at least 8 continuous residues from SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4; or 
 (iii) two or more amino acid sequences having at least 8 continuous residues from SEQ ID NO:2, SEQ ID NO:3, and/or SEQ ID NO:4. 
   
     
     
         16 . (canceled) 
     
     
         17 . A method for immunizing an individual against a cancer disease associated with positive PDEF expression, said method comprising:
 administering a vaccine to the individual, wherein said individual is at high risk for, predisposed to, susceptible to, and/or diagnosed with said cancer disease associated with positive PDEF expression, and   wherein said vaccine comprises:   (a) a viral or plasmid expression vector according to  claim 15 , and   (b) a pharmacologically acceptable carrier or adjuvant.   
     
     
         18 . The method according to  claim 17 , wherein said cancer disease is selected from the group consisting of breast cancer, prostate cancer, ovarian cancer, endometrial cancer, colon cancer, and cervical cancer. 
     
     
         19 - 27 . (canceled) 
     
     
         28 . A method of treating or preventing a disease characterized by growth of tumor cells expressing PDEF, said method comprising:
 administering to a subject a vaccine according to  claim 10 , thereby inducing T cell immunity that inhibits growth of tumor cells that express PDEF.   
     
     
         29 . A method of treating or preventing a disease characterized by growth of tumor cells expressing PDEF, said method comprising:
 administering to a subject a vaccine comprising:   (a) a viral or plasmid expression vector according to  claim 15 , and   (b) a pharmacologically acceptable carrier or adjuvant,   thereby inducing T cell immunity that inhibits growth of tumor cells that express PDEF.   
     
     
         30 . (canceled) 
     
     
         31 . (canceled) 
     
     
         32 . (canceled) 
     
     
         33 . A method for boosting immunity of a cancer patient to a cancer disease associated with positive expression of PDEF, said method comprising:
 (a) obtaining a tissue sample from the cancer patient, wherein said tissue sample is suspected of comprising cancer tumor tissue;   (b) contacting the tissue sample with the antibody or antigen-binding fragment thereof effective to allow for measuring the level of PDEF expression in the tissue sample, wherein said antibody or antigen-binding fragment thereof specifically binds with high affinity to at least a portion of a segment of a human prostate-derived Ets transcription factor (PDEF), wherein said segment comprises:
 (i) amino acid residues 1-104 of SEQ ID NO:1; 
 (ii) amino acid residues 105-141 of SEQ ID NO:1; or 
 (iii) amino acid residues 214-247 of SEQ ID NO:1; 
   (c) measuring the level of PDEF expressed in the tissue sample using an immunohistochemical assay, whereby positive expression of PDEF indicates that the patient is in need of an additional immunization against PDEF; and   (d) providing the patient with a vaccine according to  claim 10  under conditions effective to boost immunity of the cancer patient to the cancer disease associated with positive expression of PDEF.   
     
     
         34 . The method according to  claim 33 , wherein said tissue sample is selected from the group consisting of breast tissue, ovarian tissue, endometrial tissue, colon tissue, and cervical tissue. 
     
     
         35 . The method according to  claim 33 , wherein said cancer disease is selected from the group consisting of breast cancer, ovarian cancer, endometrial cancer, colon cancer, and cervical cancer.