Nucleic acid molecules encoding the MAGE-1 tumor rejection antigen precursor

Abstract

The invention relates to an isolated DNA sequence which codes for an antigen expressed by tumor cells which is recognized by cytotoxic T cells, leading to lysis of the tumor which expresses it. Also described are cells transfected by the DNA sequence, and various therapeutic and diagnostic uses arising out of the properties of the DNA and the antigen for which it codes.

1. An isolated nucleic acid molecule which codes for the MAGE tumor rejection antigen precursor as set forth in SEQ. ID. NO. 8. 
     
     
       2. The isolated nucleic acid molecule of  claim 1 , wherein said nucleic acid is a DNA cDNA molecule which comprises nucleotides 3881 to 4711 of SEQ. ID. NO. 8. 
     
     
       3. An isolated mRNA molecule which is complementary to the nucleic acid molecule of  claim 1 . 
     
     
       4. An isolated nucleic acid molecule which hybridizes to the nucleic acid molecule which codes for the MAGE-1 tumor rejection antigen precursor as set forth in SEQ ID No: 8 under stringent conditions and which codes for a tumor rejection precursor. 
     
     
       5. A expression vector comprising the isolated nucleic acid molecule of  claim 1  operably linked to a promoter. 
     
     
       6. The expression vector of  claim 5 , wherein said promoter is an incurable promoter. 
     
     
       7. An expression vector comprising the isolated nucleic acid molecule of  claim 2  operably linked to a promoter. 
     
     
       8. The expression vector of  claim 7 , wherein said promoter is an inducible promoter. 
     
     
       9. An expression vector comprising the isolated nucleic acid molecule of  claim 4  operably linked to a promoter. 
     
     
       10. The expression vector of  claim 9 , wherein said promoter is an inducible promoter. 
     
     
       11. A host cell transfected with the nucleic acid molecule of  claim 1 . 
     
     
       12. The host cell of  claim 11 , wherein said host cell is a mammalian cell. 
     
     
       13. The host cell of  claim 12 , wherein said cell line is a fibroblast cell line. 
     
     
       14. A host cell line transfected with the nucleic acid molecule of  claim 2 . 
     
     
       15. A host cell line transfected with the nucleic acid molecule of  claim 4 . 
     
     
       16. The hose cell of  claim 15 , wherein MAGE 1 has the nucleic acid sequence as follows:
                                                1   GGATCCAGGC   CCTGCCAGGA   AAAATATAAG   GGCCCTGCGT   GAGAACAGAG   GGGGTCATCC     60           61   ACTGCATGAG   AGTGGGGATG   TCACAGAGTC   CAGCCCACCC   TCCTGGTACG   ACTGAGAAGC    120      121   CAGGGCTGTG   CTTGCGGTCT   GCACCCTGAG   GGCCCGTGGA   TTCCTCTTCC   TGGAGCTCCA    180      181   GGAACCAGGC   AGTGAGGCCT   TGGTCTGAGA   CAGTATCCTC   AGGTCACAGA   GCAGAGGATG    240      241   CACAGGGTGT   GCCAGCAGTG   AATGTTTGCC   CTGAATGCAC   ACCAAGGGCC   CCACCTGCCA    300      301   CAGGACACAT   AGGACTCCAC   AGAGTCTGGC   CTCACCTCCC   TACTGTCAGT   CCTGTACAAT    360      361   CGACCTCTGC   TGGCCGGCTG   TACCCTGAGT   ACCCTCTCAC   TTCCTCCTTC   AGGTTTTCAG    420      421   GGGACAGGCC   AACCCAGAGG   ACAGGATTCC   CTGGAGGCCA   CAGAGGAGCA   CCAAGGAGAA    480      481   GATCTGTAAG   TAGGCCTTTG   TTAGAGTCTC   CAAGGTTCAG   TTCTCAGCTG   AGGCCTCTCA    540      541   CACACTCCCT   CTCTCCCCAG   GCCTGTGGGT   CTTCATTGCC   CAGCTCCTGC   CCACACTCCT    600      601   GCCTGCTGCC   CTGACGAGAG   TCATCATGTC   TCTTGAGCAG   AGGAGTCTGC   ACTGCAAGCC    660      661   TGAGGAAGCC   CTTGAGGCCC   AACAAGAGGC   CCTGGGCTGG   TGTGTGTGCA   GGCTGCCACC    720      721   TCCTCCTCCT   CTCCTCTGGT   CCTGGGCACC   CTGGAGGAGG   TGCCCACTGC   TGGGTCAACA    780      781   GATCCTCCCC   AGAGTCCTCA   GGGAGCCTCC   GCCTTTCCCA   CTACCATCAA   CTTCACTCGA    840      841   CAGAGGCAAC   CCAGTGAGGG   TTCCAGCAGC   CGTGAAGAGG   AGGGGCCAAG   CACCTCTTGT    900      901   ATCCTGGAGT   CCTTGTTCCG   AGCAGTAATC   ACTAAGAAGG   TGGCTGATTT   GGTTGGTTTT    960      961   CTGCTCCTCA   AATATCGAGC   CAGGGAGCCA   GTCACAAAGG   CAGAAATGCT   GGAGAGTGTC   1020     1021   ATCAAAAATT   ACAAGCACTG   TTTTCCTGAG   ATCTTCGGCA   AAGCCTCTGA   GTCCTTGCAG   1080     1081   CTGGTCTTTG   GCATTGACGT   GAAGGAAGCA   GACCCCACCG   GCCACTCCTA   TGTCCTTGTC   1140     1141   ACCTGCCTAG   GTCTCTCCTA   TGATGGCCTG   CTGGGTGATA   ATCAGATCAT   GCCCAAGACA   1200     1201   GGCTTCCTGA   TAATTGTCCT   GGTCATGATT   GCAATGGAGG   GCGGCCATGC   TCCTGAGGAG   1260     1261   GAAATCTGGG   AGGAGCTGAG   TGTGATGGAG   GTGTATGATG   GGAGGGAGCA   CAGTGCCTAT   1320     1321   GGGGAGCCCA   GGAAGCTGCT   CACCCAAGAT   TTGGTGCAGG   AAAAGTACCT   GGAGTACGGC   1380     1381   AGGTGCCGGA   CAGTGATCCC   GCACGCTATG   AGTTCCTGTG   GGGTCCAAGG   GCCCTCGCTG   1440     1441   AAACCAGCTA   TGTGAAAGTC   CTTGAGTATG   TGATCAAGGT   CAGTGCAAGA   GTTCGCTTTT   1500     1501   TCTTCCCATC   CCTGCGTGAA   GCAGCTTTGA   GAGAGGAGGA   AGAGGGAGTC   TGAGCATGAG   1560     1561   TTGCAGCCAA   GGCCAGTGGG   AGGGGGACTG   GGCCAGTGCA   CCTTCCAGGG   CCGCGTCCAG   1620     1621   CAGCTTCCCC   TGCCTCGTGT   GACATGAGGC   CCATTCTTCA   CTCTGAAGAG   AGCGGTCAGT   1680     1681   GTTCTCAGTA   GTAGGTTTCT   GTTCTATTGG   GTGACTTGGA   GATTTATCTT   TGTTCTCTTT   1740     1741   TGGAATTGTT   CAAATGTTTT   TTTTTAAGGG   ATGGTTGAAT   GAACTTCAGC   ATCCAAGTTT   1800     1801   ATGAATGACA   GCAGTCACAC   AGTTCTGTGT   ATATAGTTTA   AGGGTAAGAG   TCTTGTGTTT   1860     1861   TATTCAGATT   GGGAAATCCA   TTCTATTTTG   TGAATTGGGA   TAATAACAGC   AGTGGAATAA   1920     1921   GTACTTAGAA   ATGTGAAAAA   TGAGCAGTAA   AATAGATGAG   ATAAAGAACT   AAAGAAATTA   1980     1981   AGAGATAGTC   AATTCTTGCC   TTATACCTCA   GTCTATTCTG   TAAAATTTTT   AAAGATATAT   2040     2041   GCATACCTGG   ATTTCCTTGG   CTTCTTTGAG   AATGTAAGAG   AAATTAAATC   TGAATAAAGA   2100     2101   ATTCTTCCTG   TTCACTGGCT   CTTTTCTTCT   CCATGCACTG   AGCATCTGCT   TTTTGGAAGG   2160     2161   CCCTGGGTTA   GTAGTGGAGA   TGCTAAGGTA   AGCCAGACTC   ATACCCACCC   ATAGGGTCGT   2220     2221   AGAGTCTAGG   AGCTGCAGTC   ACGTAATCGA   GGTGGCAAGA   TGTCCTCTAA   AGATCTAGGG   2280     2281   AAAAGTGAGA   GAGGGGTGAG   GGTGTGGGGC   TCCGGGTGAG   AGTGGTGGAG   TGTCAATGCC   2340     2341   CTGAGCTGGG   GCATTTTGGG   CTTTGGGAAA   CTGCAGTTCC   TTCTGGGGGA   GCTGATTGTA   2400     2401   ATGATCTTGG   GTGGATCC                   2418                                                         
       
        
       
     
     
       17. The host cell of  claim 14 , wherein said host cell is a mammalian cell. 
     
     
       18. The host cell of  claim 15 , wherein said host cell is a mammalian cell. 
     
     
       19. An isolated nucleic acid molecule which hybridizes to the nucleic acid molecule which codes for the MAGE- 1  tumor rejection antigen precursor as set forth in SEQ ID NO:  8  at  0 . 1 ×SSC,  0 . 1 % SDS, and which codes for a tumor rejection antigen precursor. 
     
     
       20. An expression vector comprising the isolated nucleic acid molecule of  claim 19 , operably linked to a promoter. 
     
     
       21. The expression vector of  claim 20 , wherein said promoter is an inducible promoter. 
     
     
       22. A host cell transfected with the insolated nucleic acid molecule of  claim 19 . 
     
     
       23. The host cell of  claim 22 , wherein said isolated nucleic acid molecule has the nucleotide sequence of SEQ ID NO:  7 . 
     
     
       24. The host cell of  claim 22 , wherein said host cell is a mammalian cell. 
     
     
       25. A host cell transfected with an isolated nucleic acid molecule which hybridizes to the nucleic acid molecule which codes for the MAGE- 1  tumor rejection antigen precursor as set forth in SEQ ID NO:  8  at  0 . 1 ×SSC,  0 . 1 % SDS and which codes for a tumor rejection antigen precursor.