Prostate specific antigen oligoepitope peptide

Abstract

The invention is a prostate specific antigen oligo-epitope peptide which comprises more than one PSA epitope peptide, which conforms to one or more human HLA class I motifs. The prostate specific antigen oligo-epitope peptide in combination with various HLA-class I molecules or interactions with various T-cell receptors elicits PSA specific cellular immune responses. The prostate specific antigen oligo-epitope peptide is useful as an immunogen in the prevention or treatment of prostatic cancer, in the inhibition of prostatic cancer cells and in the establishment and characterization of PSA-specific cytotoxic T-cell lines.

1 . A prostate specific antigen oligo-epitope peptide (PSA-OP) or analog thereof comprising more than one prostate specific antigen epitope peptide adjoined, the PSA-OP generates a prostate specific response in a portion of the human population.  
     
     
         2 . The prostate specific antigen oligo-epitope peptide according to  claim 1  wherein each prostate specific antigen epitope peptide binds the same or different HLA class I molecule types.  
     
     
         3 . The prostate specific antigen oligo-epitope peptide according to  claim 1 , wherein after cleavage of PSA-OP to produce PSA-OP cleavage fragments, the PSA-OP cleavage fragments bind to one or more HLA class I molecule types.  
     
     
         4 . The prostate specific antigen oligo-epitope peptide according to  claim 3 , wherein the PSA-OP cleavage fragments are produced by a protease.  
     
     
         5 . The prostate specific antigen oligo-epitope peptide (PSA-OP) or analogs thereof according to  claim 3  wherein the PSA-OP cleavage fragments bind to one or more HLA class I molecule types selected from the group consisting of HLA-A1, HLA-A2, HLA-A3, HLA-A11, HLA-A24, HLA-A26, HLA-A28, HLA-A32, HLA-B7, HLA-B44, HLA-Cw3, HLA-Cw4, HLA-Cw5, HLA-Aw68 and HLA-B53.  
     
     
         6 . The prostate specific antigen oligo-epitope peptide or analogs thereof according to  claim 1 , wherein each prostate specific antigen epitope peptide each variably comprises an amino acid sequence of about 8 to about 12 amino acids.  
     
     
         7 . The prostate specific antigen oligo-epitope peptides or analogs thereof according to  claim 1 , wherein each prostate specific antigen epitope peptide are adjoined together directly by a peptide bond or by an amino acid linker sequence.  
     
     
         8 . The prostate specific antigen oligo-epitope peptide or analog thereof according to  claim 1 , wherein the prostate specific antigen oligo-epitope peptide comprises repeating units of one or more prostate specific antigen epitope peptides.  
     
     
         9 . The prostate specific antigen oligo-epitope peptide or analogs thereof according to  claim 1  which comprises a first prostate specific antigen epitope peptide (PSA1): 
 F L T P K K L Q C V (SEQ. ID NO.: 1) and analogs thereof and a second prostate specific antigen epitope peptide (PSA3):    V I S N D V C A Q V (SEQ. ID NO.: 2), and analogs thereof.    
     
     
         10 . The prostate specific antigen oligo-epitope peptide or analogs thereof according to  claim 9  wherein the first prostate specific antigen epitope peptide and the second prostate specific antigen epitope peptide are adjoined together directly by a peptide bond.  
     
     
         11 . The prostate specific antigen oligo-epitope peptide or analogs thereof according to  claim 9  further comprising a linker amino acid sequence which adjoins PSA1 and PSA3, the linker sequence comprising about 1 to about 10 amino acids.  
     
     
         12 . The prostate specific antigen oligo-epitope peptide or analogs thereof according to  claim 5  further comprising a third prostate specific antigen epitope peptide adjoined to the second prostate specific antigen epitope peptide.  
     
     
         13 . The prostate specific antigen oligo-epitope peptide or analogs thereof according to  claim 1  comprising:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   F    L   T   P   K   K   L   Q   C   V   D   L 
                     
                 
                     
                     
                 
                     
                   141 142 143 144 145 146 147 148 149 150 151 152 
                 
                     
                     
                 
                     
                   H    V   I   S   N   D   V   C   A   Q   V   H 
                 
                     
                   153 154 155 156 157 158 159 160 161 162 163 164 
                 
                     
                     
                 
                     
                   P    Q   K   V   T   K   (SEQ. ID. NO. 4) 
                 
                     
                   165 166 167 168 169 170 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       and analogs thereof.  
     
     
         14 . The prostate specific antigen oligo-epitope peptide or analogs thereof according to  claim 13  comprising a substitution of valine at one or more positions selected from the group consisting of 148, 149, 160 and 161.  
     
     
         15 . A pharmaceutical composition comprising the prostate specific antigen oligo-epitope peptide or analogs thereof of  claim 1  and a pharmaceutically acceptable diluent, carrier or excipient carrier.  
     
     
         16 . An isolated prostate specific antigen DNA sequence comprising a DNA sequence which encodes the prostate specific antigen oligo-epitope peptide or analogs thereof according to  claim 1 .  
     
     
         17 . An isolated prostate specific antigen DNA sequence comprising a DNA sequence which encodes the prostate specific antigen oligo-epitope peptide or analogs according to  claim 9 .  
     
     
         18 . An isolated prostate specific antigen DNA sequence comprising SEQ. ID NO.: 5, SEQ. ID NO.: 6, or analogs or variants thereof.  
     
     
         19 . A plasmid or virus vector comprising the prostate specific antigen DNA sequence according to  claim 16 .  
     
     
         20 . A virus vector comprising the DNA sequence of SEQ. ID NO.: 14.  
     
     
         21 . The vector according to  claim 19  wherein the vector is a  E. coli  plasmid, a Listeria vector, an orthopox virus, avipox virus, capripox virus, suipox virus, vaccinia virus, baculovirus, human adenovirus, SV40 or bovine papilloma.  
     
     
         22 . A host cell comprising a plasmid or virus according to  claim 19 , wherein the host cell expresses the prostate specific antigen oligo-epitope peptide or analog thereof.  
     
     
         23 . The host cell according to  claim 22  wherein the host cell binds cleavage fragments of the prostate specific antigen oligo-epitope peptide or analog thereof.  
     
     
         24 . The host cell according to  claim 23  wherein the cleavage fragments are produced by a protease.  
     
     
         25 . The host cell according to  claim 23  wherein the host cell additionally expresses a HLA class I molecule type selected from the group consisting of HLA-1, HLA-2, HLA-3, HLA-A11, HLA-A24, HLA-A26, HLA-A28, HLA-A32, HLA-B7, HLA-B44, HLA-Cw3, HLA-Cw4, HLA-Cw5, HLA-Aw68 and HLA-B53.  
     
     
         26 . A recombinant virus comprising a virus into which a prostate specific antigen (PSA) DNA sequence which encodes a prostate specific antigen oligo-epitope peptide or analog thereof according to  claim 1  is inserted, the recombinant virus causes the expression of the prostate specific antigen oligo-epitope peptide, analogs or fragments thereof in a host cell.  
     
     
         27 . A recombinant virus comprising a virus selected from the group consisting of orthopox virus, avipox virus, capripox virus, suipox virus, vaccinia virus, baculovirus, DNA plasmid, human adenovirus, SV40 and bovine papilloma into which a prostate specific antigen (PSA) DNA sequence which encodes the prostate specific antigen oligo-epitope peptide or analogs thereof is inserted, the recombinant virus causes the expression of the prostate specific antigen oligo-epitope peptide, or analogs or fragments thereof on the surface of host cells infected therewith and the infected host cells elicits an immune response directed against PSA, cells expressing PSA, cells expressing a prostate specific antigen oligo-epitope peptide or analogs thereof, or cells binding cleavage fragments of the prostate specific antigen oligo-epitope peptide or analogs thereof.  
     
     
         28 . A recombinant virus of  claim 27  further comprising a DNA sequence encoding an immunoenhancing molecule, the molecule selected from the group consisting of influenza peptide, tetanus toxoid, tetanus toxoid CD4 epitope,  Pseudomonas  exotoxin A and poly-L-lysine.  
     
     
         29 . A pharmaceutical composition comprising the recombinant virus according to  claim 27  and a pharmaceutically acceptable diluent, carrier, or excipient carrier and optionally a biological response modifier selected from the group consisting of interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-12 (IL-12) interferon, tumor necrosis factor (TNF) granulocyte monocyte-colong stimulating factor (GM-CSF) and cyclophosphamide.  
     
     
         30 . A method of stimulating the immune system of a mammal against prostate specific antigen for the purpose of preventing the establishment and growth of PSA positive carcinoma cells comprising administering to said mammal the recombinant virus according to  claim 27  in an amount sufficient to effect said stimulation.  
     
     
         31 . The method according to  claim 30  wherein the recombinant virus is vaccinia virus of the NYC strain or v-WR strain.  
     
     
         32 . The method according to  claim 31  wherein the vaccinia virus is recombined with an attenuated human vaccinia virus strain.  
     
     
         33 . The method according to  claim 30  further comprising administering with said recombinant virus a biological response modifier selected from the group consisting of interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin 12, interferon, tumor necrosis factor (TNF), GM-CSF and cyclophosphamide.  
     
     
         34 . The method according to  claim 30  further comprising administering with said recombinant virus an adjuvant.  
     
     
         35 . A method of inhibiting or killing PSA positive tumor cells comprising: 
 A) generating PSA specific cytotoxic T lymphocytes in vitro by stimulation of lymphocytes from a source with an effective amount of a prostate specific antigen oligo-epitope peptide of  claim 1 , alone or in combination with one or more cytokines, said amount is effective in generating PSA specific cytotoxic T lymphocytes; and    B) adoptively transferring the PSA specific cytotoxic T lymphocytes alone or in combination with the prostate specific antigen oligo-epitope peptide into a mammal in an amount sufficient to inhibit or kill the PSA positive tumor cells.    
     
     
         36 . A method of inhibiting or killing PSA positive tumor cells in a mammal comprising: 
 A) generating PSA specific cytotoxic T lymphocytes in vivo by administration of an effective amount of a prostate specific antigen oligo-epitope peptide of  claim 1  alone or in combination with an adjuvant or liposomes, and    B) the PSA specific cytotoxic T lymphocytes so generated inhibit or kill PSA positive tumor cells in the mammal.    
     
     
         37 . The method according to  claim 36  wherein the adjuvant is selected from the group consisting of RIBI Detox, QS21, alum and incomplete Freund's adjuvant.  
     
     
         38 . A method of generating an immune response to prostate specific antigen in a population of humans having more than one HLA-class I molecule type comprising: 
 administering of an effective amount of a prostate specific antigen oligo-epitope peptide or analogs thereof according to  claim 1 , alone or in combination with an adjuvant,    said amount is sufficient to generate a PSA specific immune response in a population of humans having more than one HLA-class I molecule type.    
     
     
         39 . The method of  claim 38  wherein the HLA-class I molecule types are selected from one or more members of the group consisting of HLA-A1, HLA-A2, HLA-A3, HLA-A11, HLA-A24, HLA-A26, HLA-A28, HLA-A32, HLA-B7, HLA-B44, HLA-Cw3, HLA-Cw4, HLA-Cw5, HLA-Aw68 and HLA-B53.  
     
     
         40 . The method of  claim 38  wherein cellular proteases from an antigen presenting cell cleaves the prostate specific antigen oligo-epitope peptide or analogs thereof into cleavage fragments which bind to one or more HLA-class I molecule types.  
     
     
         41 . The method of  claim 38 , wherein the immune response is generation and expansion of PSA specific cytotoxic T lymphocytes which kill or inhibit PSA positive cells.