Transgenic animal expressing hla-a24 and utilization thereof
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 .