TOP > 外国特許検索 > METHOD FOR CLONING T CELL RECEPTOR

METHOD FOR CLONING T CELL RECEPTOR UPDATE

外国特許コード F140007957
整理番号 S2012-0940-C0
掲載日 2014年9月26日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2013JP070028
国際公開番号 WO 2014017533
国際出願日 平成25年7月24日(2013.7.24)
国際公開日 平成26年1月30日(2014.1.30)
優先権データ
  • 特願2012-164442 (2012.7.25) JP
発明の名称 (英語) METHOD FOR CLONING T CELL RECEPTOR UPDATE
発明の概要(英語) The present invention addresses the problem of providing a T cell receptor (TCR) cloning system which enables the analysis of non-biased TCR repertories, enables the collection of an antigen-specific (TCR α cDNA)/( TCR β cDNA) pair, and also enables the evaluation of the function of the pair. Provided is a method for producing a TCR gene specific to an antigen (A), comprising the steps of: (1) stimulating a group of T cells which contains antigen-(A)-specific T cells or a single antigen-(A)-specific T cell under the conditions effective for the amplification of the TCR gene; (2) identifying the antigen-(A)-specific T cells among from the group of T cells which contains the antigen-(A)-specific T cells and then sorting the identified antigen-(A)-specific T cells into a container one by one; and (3) subjecting one of the activated antigen-(A)-specific T cells in the container to PCR to amplify the TCR gene specific to the antigen (A). According to the present invention, it becomes possible to clone a TCR gene of interest in a shorter time compared with the conventional methods, e.g., within about 10 days. According to the present invention, it also becomes possible to clone a gene for TCR α-chain and a gene for TCR β-chain with high efficiency. Under the conditions employed in examples, a (TCR α-chain)-(TCR β-chain) pair was able to be obtained with an efficiency of 100% from the T cells which had been sorted one by one after stimulation.
従来技術、競合技術の概要(英語) BACKGROUND ART
Mainly applied to a specific cancer has been studied T cell receptor (TCR) in gene therapy, lymphocytes of the cancer patient, cancer antigen-specific TCR gene is introduced. The gene transduced lymphocytes after when a large number, is returned to the cancer patient, recognizes a peptide tumor antigen was expressed on TCR since lymphocytes, cancer cells which present the tumor antigen which is recognized and specifically attacks, finally can be expected to abolish the cancer cells.
For use in gene therapy of antigen-specific TCR gene to obtain, collected from the patient in the peripheral blood lymphocytes (PBL) from the cancer antigen is expressed in cells T T cells can be identified, it is necessary to clone the gene TCR. Approach is typically performed for this purpose, including antigen-specific T cell clones established, this is typically, requires several months.
On the other hand, antigen-specific T cells for numerous studies in the TCR repertoire have been made. This is, analysis is typically done in, for example, the product of a gene family TCR β V (TRB) targeting monoclonal antibody (mAb) of panel (non-patent document 1) a method based on FACS TRBV PCR and specific primers based on the method of panel (non-patent document 2-4) has been performed by. However, conventional repertoire analysis, both TCR α TRBV V (TRA) analysis and not, as TCR repertoire analysis, is incomplete. In addition, established T cell clones TCR repertoire including these methods can cause deviations are presented the concern that the (non-patent document 5 and 6). That is, step T cell clones were established, and grow easily from T cell clones will grow and, for analysis using PCR, amplified by the primers can differ depending on the efficiency.
On the other hand, the present inventors and other groups by a group, human (non-patent document 7) and mouse (non-patent document 8) and of the transcription products of the simultaneous CDR3 α TCR CDR3 β single cell RT-PCR protocols that enable the identification have been reported. However, single cell RT-PCR protocol thereof complete protein translation region cloned TCR α / β cDNA pairs, by expressing in the cell it, in order to determine the antigen specificity has not yet been produced.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • NATIONAL UNIVERSITY CORPORATION UNIVERSITY OF TOYAMA
  • 発明者(英語)
  • KISHI, Hiroyuki
  • MURAGUCHI, Atsushi
  • KOBAYASHI, Eiji
  • OZAWA, Tatsuhiko
国際特許分類(IPC)
指定国 National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IS KE KG KN KP KR KZ LA LC LK LR LS LT LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN TD TG
ライセンスをご希望の方、特許の内容に興味を持たれた方は、下記までご連絡ください。

PAGE TOP

close
close
close
close
close
close