Cell for use in immunotherapy which contains modified nucleic acid construct encoding wilms tumor gene product or fragment thereof, method for producing said cell, and said nucleic acid construct

Abstract

A cell of the present invention contains a nucleic acid construct encoding a WT1 gene product or a fragment of the WT1 gene product. The nucleic acid construct contains (i) a region encoding a desired fragment of the WT1 gene product and (ii) only AUG as a functional start codon. The present invention can provide a cell into which the nucleic acid construct is introduced so that an expression level of a WT1 gene product or a fragment of the WT1 gene product is remarkably enhanced.

1 . A cell for immunotherapy, comprising:
 a nucleic acid construct encoding a Wilms tumor gene product or a fragment of the Wilms tumor gene product,   the nucleic acid construct including (i) a region encoding a fragment of the Wilms tumor gene product, the fragment being indicated by positions 194 to 493 of SEQ ID NO: 1 or by positions corresponding to the positions 194 to 493 of a sequence corresponding to SEQ ID NO: 1 and (ii) only one AUG as a functional start codon, connected to a 5′ terminal side of the region via 3m (m is 0 or a positive integer) bases intervening between the 5′ terminal side of the region and the AUG as the functional start codon.   
     
     
         2 . The cell as set forth in  claim 1 , wherein:
 as the region encoding the fragment of the Wilms tumor gene product, the nucleic acid construct includes a region encoding a fragment of the Wilms tumor gene product, the fragment being indicated by positions 69 to 517 of SEQ ID NO: 1 or by positions corresponding to the positions 69 to 517 of a sequence corresponding to SEQ ID NO: 1; and   the nucleic acid construct is such that all of CUG at positions 191 to 193 are deleted or C of CUG at positions 191 to 193 is substituted with A in the sequence indicated by SEQ ID NO: 2.   
     
     
         3 . The cell as set forth in  claim 1 , wherein a production amount of proteins directly translated from the nucleic acid construct is 25 times or more as large as that of proteins directly translated from RNA including a coding region indicated by positions 191 to 1741 of SEQ ID NO: 2. 
     
     
         4 . A nucleic acid construct for producing a cell recited in  claim 1 , the nucleic acid construct being directly translated into a protein in a cell into which the nucleic acid construct is introduced. 
     
     
         5 . The nucleic acid construct as set forth in  claim 4 , comprising a region indicated by SEQ ID NO: 4 or 5. 
     
     
         6 . A method for producing a cell for immunotherapy, comprising the step of introducing, into a cell, a nucleic acid construct recited in  claim 4 . 
     
     
         7 . A polynucleotide encoding a nucleic acid construct recited in  claim 4 . 
     
     
         8 . A kit for producing a cell for immunotherapy, comprising a nucleic acid construct recited in  claim 4 . 
     
     
         9 . The cell as set forth in  claim 1 , further comprising mRNA encoding CD1d, the CD1d existing on a surface of the cell and being bound to a glycolipid recognizable by an antigen receptor of an NKT cell.