Isolated nucleic acid molecules encoding cancer associated antigens, the antigens per se, and uses thereof

Abstract

The invention relates to newly identified cancer associated antigens. It has been discovered that each of these molecules provokes antibodies when expressed by a subject. The ranifications of this observation are also a part of this invention.

We claim:  
     
         1 . Isolated nucleic acid molecule which encodes a cancer associated antigen, whose amino acid sequence is identical to the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 1, 3, 4, 8, 15, 19, 22, or 26.  
     
     
         2 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleotide sequence of SEQ ID NO: 1.  
     
     
         3 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleotide sequence of SEQ ID NO: 3.  
     
     
         4 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleotide sequence of SEQ ID NO: 4.  
     
     
         5 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleotide sequence of SEQ ID NO: 8.  
     
     
         6 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleotide sequence of SEQ ID NO: 15.  
     
     
         7 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleotide sequence of SEQ ID NO: 19.  
     
     
         8 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleotide sequence of SEQ ID NO: 22.  
     
     
         9 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleotide sequence of SEQ ID NO: 26.  
     
     
         10 . Expression vector comprising the isolated nucleic acid molecule of  claim 1 , operably linked to a promoter.  
     
     
         11 . Eukaryotic cell line or prokaryotic cell strain, transformed or transfected with the expression vector of  claim 10 .  
     
     
         12 . Isolated cancer associated antigen comprising all or part of the amino acid sequence encoded by SEQ ID NO: 1, 3, 4, 8, 15, 19, 22 or 26.  
     
     
         13 . Eukaryotic cell line or prokaryotic cell strain, transformed or transfected with the isolated nucleic acid molecule of  claim 1 .  
     
     
         14 . The eukaryotic cell line or prokaryotic cell strain of  claim 13 , wherein said cell line is also transfected with a nucleic acid molecule coding for a cytokine.  
     
     
         15 . The eukaryotic cell line or prokaryotic cell strain of  claim 14 , wherein said cell line is further transfected by a nucleic acid molecule coding for an MHC molecule.  
     
     
         16 . The eukaryotic cell line or prokaryotic cell strain of  claim 14 , wherein said cytokine is an interleukin.  
     
     
         17 . The eukaryotic cell line or prokaryotic cell strain of  claim 16 , wherein said interleukin is IL-2, IL-4 or IL-12.  
     
     
         18 . The eukaryotic cell line or prokaryotic cell strain of  claim 13 , wherein said cell line has been rendered non-proliferative.  
     
     
         19 . The eukaryotic cell line of  claim 13 , wherein said cell line is a fibroblast cell line.  
     
     
         20 . Expression vector comprising a mutated or attenuated virus and the isolated nucleic acid molecule of  claim 1 .  
     
     
         21 . The expression vector of  claim 20 , wherein said virus is adenovirus or vaccinia virus.  
     
     
         22 . The expression vector of  claim 21 , wherein said virus is vaccinia virus.  
     
     
         23 . The expression vector of  claim 21 , wherein said virus is adenovirus.  
     
     
         24 . Expression system useful in transfecting a cell, comprising (i) a first vector containing a nucleic acid molecule which codes for the isolated cancer associated antigen of  claim 13  and (ii) a second vector selected from the group consisting of (a) a vector containing a nucleic acid molecule which codes for an MHC or HLA molecule which presents an antigen derived from said cancer associated antigen and (b) a vector containing a nucleic acid molecule which codes for an interleukin.  
     
     
         25 . Immunogenic composition comprising the isolated cancer antigen of  claim 12 , and a pharmaceutically acceptable adjuvant.  
     
     
         26 . The immunogenic composition of  claim 25 , wherein said adjuvant is a cytokine, a saponin, or GM-CSF.  
     
     
         27 . Immunogenic composition comprising at least one peptide consisting of an amino acid sequence of from 8 to 12 amino acids concatenated to each other in the isolated cancer associated cancer antigen of  claim 12 , and a pharmaceutically acceptable adjuvant.  
     
     
         28 . The immunogenic composition of  claim 27 , wherein said adjuvant is a saponin, a cytokine, or GM-CSF.  
     
     
         29 . The immunogenic composition of  claim 25 , wherein said composition comprises a plurality of peptides which complex with a specific MHC molecule.  
     
     
         30 . Immunogenic composition which comprises at least one expression vector which encodes a peptide derived from the amino acid sequence encoded by SEQ ID NO: 1, 3, 4, 8, 15, 19, 22 or 26.  
     
     
         31 . The immunogenic composition of  claim 30 , wherein said at least one expression vector codes for a plurality of peptides.  
     
     
         32 . Vaccine useful in treating a subject afflicted with a cancerous condition comprising the isolated eukaryotic cell line of  claim 13  and a pharmacologically acceptable adjuvant.  
     
     
         33 . The vaccine of  claim 32 , wherein said eukaryotic cell line has been rendered non-proliferative.  
     
     
         34 . The vaccine of  claim 33 , wherein said eukaryotic cell line is a human cell line.  
     
     
         35 . A composition of matter useful in treating a cancerous condition comprising a non-proliferative cell line having expressed on its surface a peptide derived from the amino acid sequence encoded by SEQ ID NO: 1, 3, 4, 8, 15, 19, 22 or 26.  
     
     
         36 . The composition of matter of  claim 35 , wherein said cell line is a human cell line.  
     
     
         37 . A composition of matter useful in treating a cancerous condition, comprising (i) a peptide derived from the amino acid sequence encoded by SEQ ID NO: 1, 3, 4, 8, 15, 19, 22 or 26, (ii) an MHC or HLA molecule, and (iii) a pharmaceutically acceptable carrier.  
     
     
         38 . Isolated antibody which is specific for the cancer associated antigen of  claim 12 .  
     
     
         39 . The isolated antibody of  claim 38 , wherein said antibody is a monoclonal antibody.  
     
     
         40 . Method for screening for cancer in a sample, comprising contacting said sample with a nucleic acid molecule which hybridizes to all or part of the molecule encoded by SEQ ID NO: 1, 2, 3, 4, 8, 15, 19, 22 or 26 and determining hybridization as an indication of cancer cells in said sample.  
     
     
         41 . A method for screening for cancer in a sample, comprising contacting said sample with the isolated antibody of  claim 38 , and determining binding of said antibody to a target as an indicator of cancer.  
     
     
         42 . Method for diagnosing a cancerous condition in a subject, comprising contacting an immune reactive cell containing sample of said subject to a cell line transfected with the isolated nucleic acid molecule of  claim 1 , and determining interaction of said transfected cell line with said immunoreactive cell said interaction being indicative of said cancer condition.  
     
     
         43 . A method for determining regression, progression of onset of a cancerous condition comprising monitoring a sample from a patient with said cancerous condition for a parameter selected from the group consisting of (i) a protein encoded by SEQ ID NO: 1, 2, 3, 4, 8, 15, 19, 22 or 26, (ii) a peptide derived from said protein, (iii) cytolytic T cells specific for said peptide and an MHC molecule with which it non-covalently complexes, and (iv) antibodies specific for said CT protein, wherein amount of said parameter is indicative of progression or regression or onset of said cancerous condition.  
     
     
         44 . The method of  claim 43 , wherein said sample is a body fluid or exudate.  
     
     
         45 . The method of  claim 43 , wherein said sample is a tissue.  
     
     
         46 . The method of  claim 43 , comprising contacting said sample with an antibody which specifically binds with said protein or peptide.  
     
     
         47 . The method of  claim 46 , wherein said antibody is labelled with a radioactive label or an enzyme.  
     
     
         48 . The method of  claim 46 , wherein said antibody is a monoclonal antibody.  
     
     
         49 . The method of  claim 43 , comprising amplifying RNA which codes for said protein.  
     
     
         50 . The method of  claim 49 , wherein said amplifying comprises carrying out polymerase chain reaction.  
     
     
         51 . The method of  claim 42 , comprising contacting said sample with a nucleic acid molecule which specifically hybridizes to a nucleic acid molecule which codes for or expresses said protein.  
     
     
         52 . The method of  claim 49 , wherein said nucleic acid molecule comprises SEQ ID NO: 9, 10, 11, 12, 13, 14, 17, 18, 20, 21, 24, 25, 28 or 29.  
     
     
         53 . The method of  claim 43 , comprising assaying said sample for shed protein.  
     
     
         54 . The method of  claim 43 , comprising assaying said sample for antibodies specific for said protein, by contacting said sample with protein.  
     
     
         55 . Method for diagnosing a cancerous condition comprising assaying a sample taken from a subject for an immunoreactive cell specific for a peptide derived from a protein encoded by SEQ ID NO: 1, 2, 3, 4, 8, 15, 19, 22 or 26, complexed to an MHC molecule, presence of said immunoreactive cell being indicative of said cancerous condition.  
     
     
         56 . Composition comprising at least one peptide consisting of an amino acid sequence of from 8 to 25 amino acids concatenated to each other in the isolated cancer associated antigen of  claim 12 , and a pharmaceutically acceptable adjuvant.  
     
     
         57 . The composition of  claim 56 , wherein said adjuvant is a saponin, a cytokine, or GM-CSF.  
     
     
         58 . The composition of  claim 56 , comprising a plurality of MHC binding peptides.  
     
     
         59 . Composition comprising an expression vector which encodes at least one peptide consisting of an amino acid sequence of from 8 to 25 amino acids concatenated to each other in the isolated cancer associated antigen of  claim 12 , and pharmaceutically acceptable adjuvant.  
     
     
         60 . The composition of  claim 59 , wherein said expression vector encodes a plurality of peptides.  
     
     
         61 . A method for screening for possible presence of a pathological condition, comprising assaying a sample from a patient believed to have a pathological condition for antibodies specific to at least one of the cancer associated antigens encoded by SEQ ID NOS: 1, 2, 3, 4, 8, 15, 19, 22 or 26, presence of said antibodies being indicative of possible presence of said pathological condition.  
     
     
         62 . The method of  claim 61 , wherein said pathological condition is cancer.  
     
     
         63 . The method of  claim 61 , wherein said cancer is melanoma.  
     
     
         64 . The method of  claim 61 , further comprising contacting said sample to purified cancer associated antigen encoded by SEQ ID NO: 1, 3, 4, 8, 15, 19, 22 or 26.  
     
     
         65 . A method for screening for possible presence of a pathological condition in a subject, comprising assaying a sample taken from said subject for expression of a nucleic acid molecule, the nucleotide sequence of which comprises SEQ ID NO: 1, 2, 3, 4, 8, 15, 19, 22 or 26, expression of said nucleic acid molecule being indicative of possible presence of said pathological condition.  
     
     
         66 . The method of  claim 65 , wherein said pathological condition is cancer.  
     
     
         67 . The method of  claim 65 , comprising determining expression via polymerase chain reaction.  
     
     
         68 . The method of  claim 65 , comprising determining expression by contacting said sample with at least one of SEQ ID NO: 9, 10, 11, 12, 13, 14, 17, 18, 20, 21, 24, 25, 28 or 29.  
     
     
         69 . A method for determining regression, progression of onset of a cancerous condition comprising monitoring a sample from a patient with said cancerous condition for a parameter selected from the group consisting of (i) a cancer associated antigen encoded by SEQ ID NO: 1, 2, 3, 4, 8, 15, 19, 22 or 25, (ii) a peptide derived from said cancer associated antigen, (iii) cytolytic T cells specific for said peptide and an MHC molecule with which it non-covalently complexes, and (iv) antibodies specific for said cancer associated antigen, wherein amount of said parameter is indicative of progression or regression or onset of said cancerous condition.  
     
     
         70 . The method of  claim 69 , wherein said sample is a body fluid or exudate.  
     
     
         71 . The method of  claim 69 , wherein said sample is a tissue.  
     
     
         72 . The method of  claim 69 , comprising contacting said sample with an antibody which specifically binds with said protein or peptide.  
     
     
         73 . The method of  claim 72 , wherein said antibody is labelled with a radioactive label or an enzyme.  
     
     
         74 . The method of  claim 72 , wherein said antibody is a monoclonal antibody.  
     
     
         75 . The method of  claim 69 , comprising amplifying RNA which codes for said protein.  
     
     
         76 . The method of  claim 75 , wherein said amplifying comprises carrying out polymerase chain reaction.  
     
     
         77 . The method of  claim 69 , comprising contacting said sample with a nucleic acid molecule which specifically hybridizes to a nucleic acid molecule which codes for or expresses said protein.  
     
     
         78 . The method of  claim 69 , comprising assaying said sample for shed cancer associated antigen.  
     
     
         79 . The method of  claim 69 , comprising assaying said sample for antibodies specific for said cancer associated antigen, by contacting said sample with said cancer associated antigen.  
     
     
         80 . Method for screening for a cancerous condition comprising assaying a sample taken from a subject for an immunoreactive cell specific for a peptide derived from a cancer associated antigen encoded by SEQ ID NO: 1, 2, 3, 4, 8, 15, 19, 22 or 26, complexed to an MHC molecule, presence of said immunoreactive cell being indicative of said cancerous condition.  
     
     
         81 . An isolated nucleic acid molecule consisting of a nucleotide sequence defined by SEQ ID NO: 1, 2, 3, 8, 15, 19, 22 or 26.  
     
     
         82 . Isolated nucleic acid molecule the complimentary sequence of which hydridizes, under stringent conditions, to the nucleotide sequence set forth in SEQ ID NO: 4, 5, 8, 15, 19, 22 or 26.  
     
     
         83 . An isolated polypeptide comprising at least 9 consecutive amino acids set forth in SEQ ID NO: 5, 7, 16, 19, 23, 27, or 30.  
     
     
         84 . The isolated polypeptide of  claim 83 , comprising at least 9 consecutive amino acids set forth in SEQ ID NO: 23 or 30.  
     
     
         85 . The isolated polypeptide of claim 84, comprising t least 9 consecutive amino acids of the amino acid sequence set forth in SEQ ID NO: 23.  
     
     
         86 . The isolated polypeptide of  claim 85 , comprising amino acids 102-111, 904-912 or 1262-1270 of SEQ ID NO: 23.  
     
     
         87 . An isolated nucleic acid molecule which encodes the amino acid sequence of SEQ ID NO: 30.  
     
     
         88 . An isolated nucleic acid molecule which encodes the isolated polypeptide of  claim 86 .  
     
     
         89 . Expression vector comprising the isolated nucleic acid molecule of  claim 88 , operably linked to a promater.