Transgenic animal expressing hla-a24 and utilization thereof

Inactive Publication Date: 2005-03-03
SUMITOMO DAINIPPON PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

0151] By returning the above therapeutic agent into the patient's body, specific CTLs are efficiently induced in the patient who is positive for both HLA-A24 and PSA, and thereby tumor can be treated. The antigen presenting cells of the present invention are especially useful for treating p

Problems solved by technology

Furthermore, it was totally unpredictable whether or not a genomic DNA of chimera HLA-A24 containing such an artificial seque

Method used

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  • Transgenic animal expressing hla-a24 and utilization thereof
  • Transgenic animal expressing hla-a24 and utilization thereof
  • Transgenic animal expressing hla-a24 and utilization thereof

Examples

Experimental program
Comparison scheme
Effect test

Example

EXAMPLE 2

Cloning of H-2K.sup.b Genomic DNA Fragment

[0169] (1) Cloning of H-2K.sup.b Genomic DNA Fragment

[0170] Mouse tumor cell line EL4 (ATCC T1B-39) was cultured, and mouse genomic DNA was purified and used in the PCR cloning. Purification of DNA was carried out using TaKaRa LA Taq.TM. (Takara Shuzo) suited for the amplification of long-chain DNA as per the attached protocol. The GenBank database was then searched for H-2K.sup.b gene needed for the construction of chimeric HLA gene, which revealed that said gene was divided in two segments registered under the Accession Nos. v00746 and v00747. As v00746, the upstream 1594 bp region of H-2K.sup.b down to midstream of intron 3 was registered and, as v00747, the downstream 1837 bp region of H-2K.sup.b down to midstream of intron 7 was registered. Because there was no BamHI restriction site in intron 3, which is divided and registered as v00746 and v00747, the H-2K.sup.b gene registered at the database was thought to have incomplete l...

Example

EXAMPLE 3

Construction of Chimera Genomic DNA (HLA-A2402 / K.sup.b DNA)

Example

[0187] The Plasmid HLA-A2402#1 containing HLA-A2402 genomic DNA obtained in Example 1 was cleaved at BglII restriction site and the plasmid H-2K.sup.b#20 / 26 containing H-2K.sup.b genomic DNA obtained in Example 2 was cleaved at BamHI restriction site, and the resultant fragments were ligated to give a recombinant plasmid. The schematic construction is shown in FIG. 2. The recombinant plasmid was introduced into E. coli JM109 (Toyobo) by heat shock method at 42.degree. C., and white colonies of E. coli to which the recombinant plasmid has been introduced were selected on ampicillin-containing LB agar medium coated with X-Gal and IPTG to obtain the transformants. Ten transformants were incubated overnight in LB medium containing ampicillin (3 ml). The plasmid clone contained in each transformant was purified and subjected to sequence analysis in a similar manner to the above. As a result, it was revealed that three transformants contained a plasmid carrying the intended chimeric gene ...

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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.

Description

[0001] The present invention relates to transgenic animals expressing HLA-A24 and utilization thereof. More specifically, the present invention relates to transgenic animals, into which an HLA-A24 gene has been introduced and in which cytotoxic T lymphocytes are induced when stimulated by an HLA-A24-binding antigen (i.e., HLA-A24-restricted antigen), a method of screening therapeutic or preventive agents for tumors or virus infections using said transgenic animals, HLA-A24-binding tumor antigen peptides of PSA origin selected by said method, recombinant DNA constructs useful in the preparation of transgenic mice and the use thereof, and the like.BACKGROUD ART[0002] The cellular immunity, particularly cytotoxic T lymphocyte (hereinafter, referred to as "CTL"), plays an important role in the removal of cancer cells or virus-affected cells from living body. CTLs recognize a complex between a peptide fragment of antigen proteins originated from cancer, virus etc. and an MHC class I mole...

Claims

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Application Information

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IPC IPC(8): A01K67/027A61P31/12A61P35/00A61P43/00C07K14/74C12N15/85
CPCA01K67/0275A01K2217/05A01K2227/105C12N15/8509A01K2267/0393C07K14/70539C07K2319/00A01K2267/03A61P31/12A61P35/00A61P43/00
Inventor GOTOH, MASASHI
Owner SUMITOMO DAINIPPON PHARMA CO LTD
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