Transgenic animal expressing hla-a24 and utilization thereof

Abstract

The present invention relates to a non-human transgenic mammal which has had an HLA-A24 gene introduced and in which CTLs are induced when stimulated by an HLA-A24-binding antigen, a method of screening therapeutic or preventive agents for tumors or virus infections comprising administering a test substance to said transgenic non-human mammal and assaying and evaluating whether CTLs specific for the test substance are induced, an HLA-A24-binding tumor antigen peptide of PSA origin selected by said screening method, a chimera DNA (DNA construct) useful in the generation of said non-human transgenic mammal, a host cell transformed by said chimera gene and use thereof, and the like.

1 . A non-human transgenic mammal, which has had an HLA-A24 gene introduced and in which CTLs are induced when stimulated by an HLA-A24-binding antigen.  
     
     
         2 . The non-human transgenic mammal according to  claim 1 , which comprises the HLA-A24 gene homozygously.  
     
     
         3 . The non-human transgenic mammal according to  claim 1  or  2 , wherein the HLA-A24 gene is a chimera gene comprising the α1 and α2 domains of HLA-A24 gene and the α3 domain of mouse MHC class I gene.  
     
     
         4 . The non-human transgenic mammal according to any one of claims  1 - 3 , wherein the HLA-A24 gene is HLA-A2402 gene.  
     
     
         5 . The non-human transgenic mammal according to any one of claims  1 - 4 , wherein the HLA-A24 gene comprises a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 3.  
     
     
         6 . The non-human transgenic mammal according to any one of claims  1 - 4 , wherein the HLA-A24 gene comprises the nucleotide sequence shown in SEQ ID NO: 2.  
     
     
         7 . The non-human transgenic mammal according to any one of claims  1 - 4 , wherein the HLA-A24 gene comprises a nucleotide sequence shown in SEQ ID NO: 1.  
     
     
         8 . The non-human transgenic mammal according to any one of claims  1 - 4 , wherein the HLA-A24 gene is a DNA which hybridizes to the complement of a nucleotide sequence set forth in any one of claims  5 - 7  under stringent conditions, and the expression product of said DNA is capable of binding to HLA-A24-binding antigen peptide and inducing CTLs.  
     
     
         9 . The non-human transgenic mammal according to any one of claims  1 - 8 , wherein the non-human mammal is mouse.  
     
     
         10 . The non-human transgenic mammal according to  claim 9  wherein the mouse is of C57BL/6 mouse strain.  
     
     
         11 . A method of screening an agent inducing antigen-specific CTLs, which comprises administering a test substance to a transgenic non-human mammal set forth in any one of claims  1 - 10 , and assaying and determining whether CTLs specific for the test substance are induced.  
     
     
         12 . A method of screening a therapeutic or preventive agent for tumors or virus infections, which comprises administering a test substance to a transgenic non-human mammal set forth in any one of claims  1 - 10 , and assaying and determining whether CTLs specific for the test substance are induced.  
     
     
         13 . The method of screening according to  claim 11  or  12 , which uses transformants transformed by and expressing the HLA-A24 gene that the transgenic non-human mammal set forth in any one of claims  1 - 10  contains as a target cell for assaying and determining whether CTLs specific for the test substance are induced.  
     
     
         14 . A transformant transformed by and expressing the HLA-A24 gene that the transgenic non-human mammal set forth in any one of claims  3 - 10  contains.  
     
     
         15 . A method of screening an antigen-specific CTL inducing agent, which uses the transformant set forth in  claim 14 .  
     
     
         16 . A method of screening a therapeutic or preventive agent for tumors or virus infections, which uses the transformant set forth in  claim 14 .  
     
     
         17 . An isolated DNA comprising the nucleotide sequence shown in SEQ ID NO: 1.  
     
     
         18 . An isolated DNA comprising the nucleotide sequence shown in SEQ ID NO: 2.  
     
     
         19 . An isolated DNA comprising the nucleotide sequence shown in SEQ ID NO: 3.  
     
     
         20 . An isolated DNA, which hybridizes to the complement of a DNA set forth in any one of claims  17 - 19  under stringent conditions, and of which expression product is capable of binding to an HLA-A24-binding antigen peptide and inducing CTLs.  
     
     
         21 . The isolated DNA according to  claim 20 , which comprises nucleotide sequences each encoding amino acid Nos. 25-114, 115-206 and 207-298 of the amino acid sequence shown in SEQ ID NO: 3.  
     
     
         22 . The isolated DNA according to  claim 20 , which rises polynucleotides each having the sequences corresponding to nucleotide Nos. 72-339, 342-615 and 618-891 of the nucleotide sequence shown in SEQ ID NO: 2.  
     
     
         23 . An expression vector comprising a DNA set forth in any one of claims  17 - 22 .  
     
     
         24 . A transformant transformed by the expression vector set forth in  claim 23 .  
     
     
         25 . A method of screening an antigen-specific CTL inducing agent, which uses the transformant set fort in  claim 24 .  
     
     
         26 . A method of screening a therapeutic or preventive agent for tumors or virus infections, which uses the transformant set fort in  claim 24 .  
     
     
         27 . An HLA-A24-binding tumor antigen peptide of PSA origin, which is obtainable by the screening method set fort in any one of claims  11 - 13 , or a derivative thereof having functionally equivalent characteristics.  
     
     
         28 . The tumor antigen peptide according to  claim 27 , which comprises the amino acid sequence shown in SEQ ID NO: 15, or a derivative thereof having functionally equivalent characteristics.  
     
     
         29 . The tumor antigen peptide derivative according to  claim 28 , which comprises the amino acid sequence shown in SEQ ID NO: 17.  
     
     
         30 . A CTL inducing agent comprising as an active ingredient the tumor antigen peptide or a derivative thereof set forth in any one of claims  27 - 29 .  
     
     
         31 . A DNA encoding a tumor antigen peptide or a derivative thereof set forth in any one of claims  27 - 29 .  
     
     
         32 . A recombinant DNA comprising the DNA set forth in  claim 31 .  
     
     
         33 . A polypeptide obtainable by expressing the recombinant DNA set forth in  claim 32 .  
     
     
         34 . A CTL inducing agent comprising as an active ingredient the recombinant DNA set forth in  claim 32  or a polypeptide set forth in  claim 33 .  
     
     
         35 . An antigen presenting cell presenting a complex between an HLA-A24 antigen and a tumor antigen peptide or a derivative thereof set forth in any one of claims  27 - 29 .  
     
     
         36 . A CTL inducing agent comprising as an active ingredient the antigen-presenting cell set forth in  claim 35 .  
     
     
         37 . A CTL which recognizes a complex between an HLA-A24 antigen and a tumor antigen peptide or a derivative thereof set forth in any one of claims  27 - 29 .  
     
     
         38 . A therapeutic or preventive agent for tumors comprising as an active ingredient the tumor antigen peptide or a derivative thereof set forth in any one of claims  27 - 29 , the recombinant DNA set forth in  claim 32 , the polypeptide set forth in  claim 33 , the antigen presenting cell set forth in  claim 35  or the CTL set forth in  claim 37 .  
     
     
         39 . A diagnosing agent comprising a tumor antigen peptide or a derivative thereof set forth in any one of claims  27 - 29 .