JP2020532959A - T細胞の改変 - Google Patents
T細胞の改変 Download PDFInfo
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- JP2020532959A JP2020532959A JP2020508395A JP2020508395A JP2020532959A JP 2020532959 A JP2020532959 A JP 2020532959A JP 2020508395 A JP2020508395 A JP 2020508395A JP 2020508395 A JP2020508395 A JP 2020508395A JP 2020532959 A JP2020532959 A JP 2020532959A
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Abstract
Description
(i)生理活性分子をコードする第1のヌクレオチド配列と、
(ii)抗原受容体をコードする第2のヌクレオチド配列と、
(iii)第1のヌクレオチド配列に作動可能に連結された誘導性プロモーターと、
(iv)第2のヌクレオチドに作動可能に連結された構成的プロモーターと、を含む、核酸構築物を提供する。
第1または第2の態様による核酸構築物またはベクターを、ドナー個体から得られたT細胞集団に導入して改変T細胞集団を生成すること、を含む、方法を提供する。
第1または第2の態様による核酸構築物またはベクターを、ドナー個体から得られたT細胞集団に導入して改変T細胞集団を生成することと、
改変T細胞集団をレシピエント個体に投与することと、を含む、方法を提供する。
本発明は、IL−7等の生理活性分子を誘導性に発現し、抗原受容体を構成的に発現する改変T細胞に関する。改変T細胞は、
(i)生理活性分子をコードする第1のヌクレオチド配列と、
(ii)抗原受容体をコードする第2のヌクレオチド配列と、
(iii)第1のヌクレオチド配列に作動可能に連結された誘導性プロモーターと、
(iv)第2のヌクレオチドに作動可能に連結された構成的プロモーターと、を含む、核酸構築物を含んでもよい。
癌を有する個体から癌細胞の試料を得ることと、
該癌細胞が、第2のヌクレオチド配列によってコードされ、改変T細胞によって発現される抗原受容体に結合すると同定することと、を含む方法によって同定され得る。
癌を有する個体から癌細胞の試料を得ることと、
該癌細胞に特異的に結合する抗原受容体を同定することと、によって同定され得る。
1.方法
1.1.IL−7発現構築物の設計および生成
同族ペプチドMHC1複合体の認識後にIL−7の誘導性発現を可能にするレンチウイルス発現ベクターを設計した。誘導性エレメントは、ヒトIL2プロモーターに存在するNFAT/AP1転写応答エレメント(TRE)に基づいていた(Macian et al.,2001 Oncogene)。IL−7の選択的な誘導性発現は、TCR活性化と、NFAT TREを内部に有する遺伝子の転写を促進することができるNFATのCa2+依存性核内移行の結果である。
1.赤色で強調表示されたヒトIL2プロモーターNFAT TREの3つのタンデム反復配列(GGAGGAAAAACTGTTTCATACAGAAGGCGT)
2.最小CMVプロモーター(緑色)
3.Kozak配列(GCCGCCACCATG)およびコドン最適化IL−7コード配列(黄色)。補足図1は野生型ヌクレオチド配列を含む。
4.SV40ポリアデニル化シグナル(紫色)
EcoRV制限部位(図1中、緑色で強調表示)の上流の配列は、レンチウイルスベクターのバックボーン配列に対応し、EF1Aプロモーターのコード配列を維持するために合成構築物中に含まれた。必要に応じて構築物のさらなる操作を可能にするために、追加の固有の制限部位も合成構築物の設計に組み入れられた。それらは以下を含む:
1.NFATIL−2 TREの上流のEcoRV(GATATC)およびNSI−I(ATGCAT)制限部位
2.IL−7CDSのすぐ上流のNdeI制限部位CATATG
3.IL−7CDSのTGA停止コドンの直後のXhoI(CTCGAG)
4.mCMVプロモーターのすぐ上流のAaTiI制限部位(GACGTC)。
NFAT_IL−7カセット(配列番号1)を、5’AgeI(ACCGGT)3’MluI(ACGCGT)制限部位を用いてGeneArt(Thermo Fisher Scientific)により人工的に合成し、標準的なクローニングベクター(pMS−RQ)中に提供した(図2)。ベクターおよびADB934(TCR1)を含むNFAT_IL−7をAgeIおよびMluIで消化し、該当する断片を製造業者の指示に従ってNucleoSpin(登録商標)ゲル抽出キットを使用してゲル抽出により精製した。消化された断片を、標準プロトコルに従ってT4 DNAリガーゼ(NEB)を用いてライゲーションした。クローンを制限酵素消化によりスクリーニングし、シークエンシングにより検証した(図4のベクターマップを参照)。
IL−7_T2A_TCR1インサートを重複PCRにより作製した。IL−7CDSは、プライマーNHEI_IL_7FおよびIL7_T2ARを用いて増幅した。TCR1 CDSは、プライマーT2AFおよびBETA_SAL_REVIIIを用いて増幅した。TCR1 CDSは、既存のインハウスの構築物ADB951から増幅した。標準プロトコルに従ってQ5 DNAポリメラーゼ(NEB)を用いてPCR反応を行い、PCR産物を製造業者の指示に従ってNucleoSpin(登録商標)ゲル抽出キットを使用してゲル抽出により精製した。NheI(GCTAGC)およびSalI(GTCGAC)制限部位(下線)を含むNHEI_IL_7FおよびBETA_SAL_REVIIIプライマーを用いて重複PCRにより2つの断片を一緒に融合した。消化された断片を、標準プロトコルに従ってT4 DNAリガーゼ(NEB)を用いてライゲーションした。クローンを制限酵素消化によりスクリーニングし、シークエンシングにより検証した(図5のベクターマップを参照)。
TCR2インサートをインハウスの構築物ADB1535からADB1224にサブクローニングした。ABD1535およびADB1224をNheIおよびSalI制限酵素で消化し、該当する断片を製造業者の指示に従ってNucleoSpin(登録商標)ゲル抽出キットを使用してゲル抽出により精製した。消化された断片を、標準プロトコルに従ってT4 DNAリガーゼ(NEB)を用いてライゲーションした。NFAT IL−7カセットを、ADB967(NFAT−IL−7_TCR1)から、AgeI制限部位とMluI制限部位の間のこの新しいTCR2レンチウイルス発現ベクターにサブクローニングした。クローンを制限酵素消化によりスクリーニングし、シークエンシングにより検証した(図6のベクターマップを参照)。
以下のプライマーを用いてIL−7 CDSをABD1099からPCR増幅した:順方向−5’TAATGCTAGCGCCGCCACCATGTTCCACGTGTCC3’および逆方向5’−CTTCACGGGCGCGCCAGAGCCGCTTCTCTTGG−3’。これらは、NheI(GCTAGC)およびAsCi(GGCGCGCC)制限部位(下線)を含む。PCR産物を製造業者の指示に従ってNucleoSpin(登録商標)ゲル抽出キットを使用してゲル抽出により精製した。PCR産物および(ADB1488)をNheIおよびASCIで消化し、該当する断片を製造業者のプロトコルに従ってNucleoSpin(登録商標)ゲル抽出キットを使用してゲル抽出により精製した。消化された断片を、標準プロトコルに従ってT4 DNAリガーゼ(NEB)を用いてライゲーションした。クローンを制限酵素消化によりスクリーニングし、シークエンシングにより検証した(図7のベクターマップを参照)。
6人のドナーからのPBLを、MACS単離キットおよび関連する製造業者のプロトコルを使用して、追加的なCD14枯渇(PBL)により全血から分離した。以下を含む異なるレンチウイルス構築物でT細胞を形質導入した:TCRおよびNFAT_IL−7 TCR(図1を参照)。T細胞の形質導入は、1mg/MlのF108ポロクサマーの存在下で行った。F108ポロクサマーは、100mg/mlのストック溶液を作製するために予め水に溶解しておき、次いで0.2μmフィルターを使用して滅菌した後、4℃で保存した。レンチウイルス粒子の添加と同時に、各ウェルの細胞(NTD細胞を含む)に1mg/MlのF108ポロクサマーを加えた。
サイトカイン産生アッセイを、典型的には以下のように行った。アッセイ当日、A375、Mel624およびColo205細胞を採取し、R10に再懸濁し、計数した。A375細胞の一部を10μM MAGE−A4ペプチドを用いて37℃/5% CO2で2時間パルスした後、3回洗浄した。標的細胞は、典型的には、3通りのウェルの96ウェル平底プレートに、100μlの体積中50,000細胞/ウェルで播種した。T細胞を解凍し、洗浄し、1×106細胞/ml未満でR10に再懸濁し、37℃/5% CO2で1〜2時間静置した。非形質導入および形質導入エフェクター細胞を計数し、3通りのウェルに、100μlのアッセイ培地にウェル当たり120,000細胞で播種し、200μlの最終体積を得た。注:異なる構築物間の形質導入の差異を考慮に入れるために、同じドナーからのNTD T細胞を加えることにより細胞を正規化し、同等の形質導入効率を得た。次いで、プレートを37℃/5% CO2で48時間インキュベーターに入れた。48時間後、アッセイプレートを遠心分離し、上清を新しい96ウェルプレートに移し、後日ELISAを行うまで−20℃で保存した。
このアッセイは、変更点を伴う製造業者のプロトコルに従ってDuoset Human IL−7 ELISAキットを使用して、96ウェルハーフエリアプレートで行った。簡潔に述べると、96ウェルハーフエリアプレートを使用したため、プロトコルに示される100μlの代わりに、全ての試薬を25μl/ウェル(または50μlの遮断試薬)で使用した。ほとんどの場合、試料は無希釈で使用した。希釈した場合、それらはアッセイ培地中に希釈された。市販のTMB基質溶液を約10分間使用してアッセイを展開し、1N H2SO4で反応を停止した。波長補正を540nmに設定して、Spectrastar Omegaを450nmで使用してアッセイを読み取った。Spectrastarデータ解析ソフトウェアを使用してデータを分析した。標準曲線上の最大濃度を超えるデータ点には、通常、最大濃度の値を割り当てた。ほとんどのアッセイで、製造業者によって示されるよりも広い範囲を提供するように標準曲線を延長し、最大濃度は1000pg/mlであった。試料および標準曲線は、2通りまたは3通りのいずれかのウェルを使用して行い、曲線のあてはめは、可能であれば、4パラメータロジスティック曲線のあてはめを用いて行った。
アッセイ当日に、T細胞を解凍し、PBS中で2回洗浄し、計数し、PBS中1×106細胞/mlに調節した。T細胞のフローサイトメトリーを行った。使用した抗体のリストは、表1および表2に見ることができる。
再刺激アッセイを以下のように行った。アッセイの初日(0日目)にT細胞を解凍し、洗浄し、1×106細胞/ml未満でR10に再懸濁し、静置した。細胞を計数し、異なるレンチウイルス調製物間の形質導入の差異を考慮に入れるために、同じドナーからのNTD T細胞を加えることにより細胞を正規化し、ドナーT細胞のバッチ内で同等の形質導入効率を得た。
1.細胞数の倍数変化=再刺激後の細胞数(28日目)/再刺激のために播種した細胞数(21日目に最初に播種した8×106個)
2.28日目に再計算した総細胞数=細胞数の倍数変化(21〜28日目)×21日目に播種するために使用可能な細胞の総数(例えば、20×106個)
A375標的細胞を採取し、48Gyで照射してそれらの増殖を阻止し、次いで、R10中に約1×106/ml、1ml/ウェル(1:1のT細胞:A375)でT細胞に加えた。対照ウェルにおいて、20μg/mlの最終濃度で組換えヒトIL−7を細胞に加えた(ウェル内の全てのIL−7が「消費された」と仮定)。この再刺激プロセスを、7、14および21日目に繰り返した。
LeukopakからのPBLは、以前に記載した方法を用いてCD14を枯渇させた。T細胞を以前に記載したように形質導入し、14日間拡大培養した後で凍結させた。NTD T細胞、およびTCR1、NFAT−IL−7_TCR1または構成的IL−7_TCR1を発現する構築物で形質導入されたT細胞に加えて、NFAT−IL−7_無関係TCRを発現する構築物でもT細胞を形質導入した。
誘導性IL−7を操作されたTCRと共発現するT細胞を注射したマウスは、TCRを単独でまたはTCRを構成的IL−7とともに発現する細胞と比較して改善された生存率を示すことが分かった(図16)。
配列
ACCGGTTCAATTGCCGACCCCTCCCCCCAACTTCTCGGGGACTGTGGGCGATGTGCGCTCTGCCCACTGACGGGCACCGGAGCCTCACGATGCATGATATCGGCCTAACTGGCCGGTACCTGAGCTCGCTAGCGGAGGAAAAACTGTTTCATACAGAAGGCGTGGAGGAAAAACTGTTTCATACAGAAGGCGTGGAGGAAAAACTGTTTCATACAGAAGGCGTgacgtcTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCAGCCTCTCGACATTCGTTGGATCCATATGGCCGCCACCATGTTCCATGTTTCTTTTAGGTATATCTTTGGACTTCCTCCCCTGATCCTTGTTCTGTTGCCAGTAGCATCATCTGATTGTGATATTGAAGGTAAAGATGGCAAACAATATGAGAGTGTTCTAATGGTCAGCATCGATCAATTATTGGACAGCATGAAAGAAATTGGTAGCAATTGCCTGAATAATGAATTTAACTTTTTTAAAAGACATATCTGTGATGCTAATAAGGAAGGTATGTTTTTATTCCGTGCTGCTCGCAAGTTGAGGCAATTTCTTAAAATGAATAGCACTGGTGATTTTGATCTCCACTTATTAAAAGTTTCAGAAGGCACAACAATACTGTTGAACTGCACTGGCCAGGTTAAAGGAAGAAAACCAGCTGCCCTGGGTGAAGCCCAACCAACAAAGAGTTTGGAAGAAAATAAATCTTTAAAGGAACAGAAAAAACTGAATGACTTGTGTTTCCTAAAGAGACTATTACAAGAGATAAAAACTTGTTGGAATAAAATTTTGATGGGCACTAAAGAACACTGACTCGAGAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTAACGCGT
配列番号1 NFAT IL−7誘導性カセットのヌクレオチド配列。
キー:
赤色(完全下線)=ヒトIL2プロモーターNFAT TREのタンデム反復配列(GGAGGAAAAACTGTTTCATACAGAAGGCGT)。
緑色(破線)=最小CMVプロモーター。
黒色=Kozak配列(GCCGCCACCATG)およびコドン最適化IL−7コード配列。
青色(点線下線)=SV40ポリアデニル化シグナル。
atgttccatgtttcttttaggtatatctttggacttcctcccctgatccttgttctgttg
M F H V S F R Y I F G L P P L I L V L L
ccagtagcatcatctgattgtgatattgaaggtaaagatggcaaacaatatgagagtgtt
P V A S S D C D I E G K D G K Q Y E S V
ctaatggtcagcatcgatcaattattggacagcatgaaagaaattggtagcaattgcctg
L M V S I D Q L L D S M K E I G S N C L
aataatgaatttaacttttttaaaagacatatctgtgatgctaataaggaaggtatgttt
N N E F N F F K R H I C D A N K E G M F
ttattccgtgctgctcgcaagttgaggcaatttcttaaaatgaatagcactggtgatttt
L F R A A R K L R Q F L K M N S T G D F
gatctccacttattaaaagtttcagaaggcacaacaatactgttgaactgcactggccag
D L H L L K V S E G T T I L L N C T G Q
gttaaaggaagaaaaccagctgccctgggtgaagcccaaccaacaaagagtttggaagaa
V K G R K P A A L G E A Q P T K S L E E
aataaatctttaaaggaacagaaaaaactgaatgacttgtgtttcctaaagagactatta
N K S L K E Q K K L N D L C F L K R L L
caagagataaaaacttgttggaataaaattttgatgggcactaaagaacactga
Q E I K T C W N K I L M G T K E H −
コドン最適化IL−7の翻訳されたヌクレオチド配列(配列番号2)およびタンパク質配列(配列番号3)
MFHVSFRYIFGLPPLILVLLPVASSDCDIEGKDGKQYESVLMVSIDQLLDSMKEIGSNCLNNEFNFFKRHICDANKEGMFLFRAARKLRQFLKMNSTGDFDLHLLKVSEGTTILLNCTGQVKGRKPAALGEAQPTKSLEENKSLKEQKKLNDLCFLKRLLQEIKTCWNKILMGTKEH*
配列番号3 IL−7タンパク質配列
ATGGAGACCCTGCTGGGCCTGCTGATCCTGTGGCTGCAGCTCCAGTGGGTGTCCAGCAAGCAGGAGGTGACCCAGATCCCTGCCGCCCTGAGCGTGCCCGAGGGCGAGAACCTGGTGCTGAACTGCAGCTTCACCGACTCCGCCATCTACAACCTGCAGTGGTTCCGGCAGGACCCCGGCAAGGGCCTGACCAGCCTGCTGCTGATCCAGAGCAGCCAGCGGGAGCAGACCAGCGGACGGCTGAACGCCAGCCTGGACAAGAGCAGCGGCCGGAGCACCCTGTACATCGCCGCCAGCCAGCCCGGCGACAGCGCCACCTACCTGTGCGCTGTGCGGCCTCTGTACGGCGGCAGCTACATCCCCACCTTCGGCAGAGGCACCAGCCTGATCGTGCACCCCTACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGTCTGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAATGTGAGCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGCTGCGACGTGAAACTGGTGGAGAAGAGCTTCGAGACCGACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCAGAATCCTGCTGCTGAAGGTGGCCGGATTCAACCTGCTGATGACCCTGCGGCTGTGGAGCAGCGGCTCCCGGGCCAAGAGAAGCGGATCCGGCGCCACCAACTTCAGCCTGCTGAAGCAGGCCGGAGACGTGGAAGAAAACCCTGGCCCTAGGATGAGCATCGGCCTGCTGTGCTGCGCCGCCCTGAGCCTGCTGTGGGCAGGACCCGTGAACGCCGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAAACCGGCCAGAGCATGACCCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTATCGGCAGGACCCCGGCATGGGCCTGCGGCTGATCCACTACTCTGTGGGAGCCGGAATCACCGACCAGGGCGAGGTGCCCAACGGCTACAATGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCTGCCCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCTATGTGGGCAACACCGGCGAGCTGTTCTTCGGCGAGGGCTCCAGGCTGACCGTGCTGGAGGACCTGAAGAACGTGTTCCCCCCCGAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACACTGGTGTGTCTGGCCACCGGCTTCTACCCCGACCACGTGGAGCTGTCCTGGTGGGTGAACGGCAAGGAGGTGCACAGCGGCGTGTCTACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGACAGCCGGTACTGCCTGTCCTCCAGACTGAGAGTGAGCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCCGGTGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATTGTGAGCGCCGAGGCCTGGGGCAGGGCCGACTGCGGCTTCACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCTGCCCTGGTGCTGATGGCTATGGTGAAGCGGAAGGACAGCCGGGGCTAAGTCGAC
配列番号4 NheI部位とSalI部位の間のヌクレオチド配列
METLLGLLILWLQLQWVSSKQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLTSLLLIQSSQREQTSGRLNASLDKSSGRSTLYIAASQPGDSATYLCAVRPLYGGSYIPTFGRGTSLIVHPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSSGSRAKRSGSGATNFSLLKQAGDVEENPGPRMSIGLLCCAALSLLWAGPVNAGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFFGEGSRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG*V
配列番号5 TCR1のタンパク質配列
METLLGLLILWLQLQWVSSKQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLTSLLLIQSSQREQTSGRLNASLDKSSGRSTLYIAASQPGDSATYLCAVRPLYGGSYIPTFGRGTSLIVHPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSSGSRAKRSGSGATNFSLLKQAGDVEENPGPRMSIGLLCCAALSLLWAGPVNAGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFFGEGSRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG
配列番号6 TCR1 CDS
ACGCGTTAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTCTCGAGTCAGTGCTCTTTGGTGCCCATCAGGATCTTGTTCCAGCAGGTCTTGATTTCCTGCAGCAGCCGCTTCAGGAAGCACAGGTCGTTCAGTTTCTTCTGCTCTTTCAGGGACTTGTTCTCTTCCAGGCTCTTGGTAGGCTGGGCTTCTCCCAGGGCGGCAGGCTTTCTGCCCTTCACTTGGCCGGTGCAATTCAGCAGGATGGTGGTGCCCTCGGACACTTTCAGCAGATGCAGGTCGAAGTCGCCGGTGCTGTTCATCTTCAGGAACTGCCGCAGCTTTCTGGCGGCTCTGAACAGGAACATGCCTTCTTTGTTGGCGTCGCAGATGTGCCGCTTGAAGAAGTTGAACTCGTTGTTCAGGCAGTTGCTGCCGATTTCCTTCATGCTGTCCAGCAGCTGGTCGATGGACACCATCAGCACGCTCTCGTACTGCTTGCCGTCCTTGCCCTCGATGTCGCAGTCGCTGCTGGCCACAGGCAGCAGCACCAGGATCAGGGGGGGCAGGCCGAAGATGTACCGGAAGGACACGTGGAACATGGTGGCGGCCATATGGATCCAACGAATGTCGAGAGGCTGGATCGGTCCCGGTGTCTTCTATGGAGGTCAAAACAGCGTGGATGGCGTCTCCAGGCGATCTGACGGTTCACTAAACGAGCTCTGCTTATATAGGCCTCCCACCGTACACGCCTAGACGTCACGCCTTCTGTATGAAACAGTTTTTCCTCCACGCCTTCTGTATGAAACAGTTTTTCCTCCACGCCTTCTGTATGAAACAGTTTTTCCTCCGCTAGCGAGCTCAGGTACCGGCCAGTTAGGCCGATATCATGCATCGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCTGCAGTACGTGATTCTTGATCCCGAGCTTCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGCTTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCCTGGGCGCTGGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGATGACCTGCTGCGACGCTTTTTTTCTGGCAAGATAGTCTTGTAAATGCGGGCCAAGATCTGCACACTGGTATTTCGGTTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTGGCCGGCCTGCTCTGGTGCCTGGCCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTGCAGGGAGCTCAAAATGGAGGACGCGGCGCTCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAGGAAAAGGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACTGAGTACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGTGCTTTTGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTGGGTGGAGACTGAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCTTTTTGAGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAGGTGTCGTGAGCTAGCCGCCACCATGGAGACCCTGCTGGGCCTGCTGATCCTGTGGCTGCAGCTCCAGTGGGTGTCCAGCAAGCAGGAGGTGACCCAGATCCCTGCCGCCCTGAGCGTGCCCGAGGGCGAGAACCTGGTGCTGAACTGCAGCTTCACCGACTCCGCCATCTACAACCTGCAGTGGTTCCGGCAGGACCCCGGCAAGGGCCTGACCAGCCTGCTGCTGATCCAGAGCAGCCAGCGGGAGCAGACCAGCGGACGGCTGAACGCCAGCCTGGACAAGAGCAGCGGCCGGAGCACCCTGTACATCGCCGCCAGCCAGCCCGGCGACAGCGCCACCTACCTGTGCGCTGTGCGGCCTCTGTACGGCGGCAGCTACATCCCCACCTTCGGCAGAGGCACCAGCCTGATCGTGCACCCCTACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGTCTGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAATGTGAGCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGCTGCGACGTGAAACTGGTGGAGAAGAGCTTCGAGACCGACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCAGAATCCTGCTGCTGAAGGTGGCCGGATTCAACCTGCTGATGACCCTGCGGCTGTGGAGCAGCGGCTCCCGGGCCAAGAGAAGCGGATCCGGCGCCACCAACTTCAGCCTGCTGAAGCAGGCCGGAGACGTGGAAGAAAACCCTGGCCCTAGGATGAGCATCGGCCTGCTGTGCTGCGCCGCCCTGAGCCTGCTGTGGGCAGGACCCGTGAACGCCGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAAACCGGCCAGAGCATGACCCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTATCGGCAGGACCCCGGCATGGGCCTGCGGCTGATCCACTACTCTGTGGGAGCCGGAATCACCGACCAGGGCGAGGTGCCCAACGGCTACAATGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCTGCCCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCTATGTGGGCAACACCGGCGAGCTGTTCTTCGGCGAGGGCTCCAGGCTGACCGTGCTGGAGGACCTGAAGAACGTGTTCCCCCCCGAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACACTGGTGTGTCTGGCCACCGGCTTCTACCCCGACCACGTGGAGCTGTCCTGGTGGGTGAACGGCAAGGAGGTGCACAGCGGCGTGTCTACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGACAGCCGGTACTGCCTGTCCTCCAGACTGAGAGTGAGCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCCGGTGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATTGTGAGCGCCGAGGCCTGGGGCAGGGCCGACTGCGGCTTCACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCTGCCCTGGTGCTGATGGCTATGGTGAAGCGGAAGGACAGCCGGGGCTAAGTCGAC
配列番号7 MluI制限部位とSalI制限部位の間の領域に及ぶセンス鎖5’から3’配列。
GCTAGCGCCGCCACCATGTTCCACGTGTCCTTCCGGTACATCTTCGGCCTGCCCCCCCTGATCCTGGTGCTGCTGCCTGTGGCCAGCAGCGACTGCGACATCGAGGGCAAGGACGGCAAGCAGTACGAGAGCGTGCTGATGGTGTCCATCGACCAGCTGCTGGACAGCATGAAGGAAATCGGCAGCAACTGCCTGAACAACGAGTTCAACTTCTTCAAGCGGCACATCTGCGACGCCAACAAAGAAGGCATGTTCCTGTTCAGAGCCGCCAGAAAGCTGCGGCAGTTCCTGAAGATGAACAGCACCGGCGACTTCGACCTGCATCTGCTGAAAGTGTCCGAGGGCACCACCATCCTGCTGAATTGCACCGGCCAAGTGAAGGGCAGAAAGCCTGCCGCCCTGGGAGAAGCCCAGCCTACCAAGAGCCTGGAAGAGAACAAGTCCCTGAAAGAGCAGAAGAAACTGAACGACCTGTGCTTCCTGAAGCGGCTGCTGCAGGAAATCAAGACCTGCTGGAACAAGATCCTGATGGGCACCAAAGAGCACGGAAGCAGAGCCAAGAGAAGCGGCTCTGGCGAGGGCAGAGGCAGCCTGCTGACATGTGGCGACGTGGAAGAAAACCCTGGCCCTATGGAGACCCTGCTGGGCCTGCTGATCCTGTGGCTGCAGCTCCAGTGGGTGTCCAGCAAGCAGGAGGTGACCCAGATCCCTGCCGCCCTGAGCGTGCCCGAGGGCGAGAACCTGGTGCTGAACTGCAGCTTCACCGACTCCGCCATCTACAACCTGCAGTGGTTCCGGCAGGACCCCGGCAAGGGCCTGACCAGCCTGCTGCTGATCCAGAGCAGCCAGCGGGAGCAGACCAGCGGACGGCTGAACGCCAGCCTGGACAAGAGCAGCGGCCGGAGCACCCTGTACATCGCCGCCAGCCAGCCCGGCGACAGCGCCACCTACCTGTGCGCTGTGCGGCCTCTGTACGGCGGCAGCTACATCCCCACCTTCGGCAGAGGCACCAGCCTGATCGTGCACCCCTACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGTCTGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAATGTGAGCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGCTGCGACGTGAAACTGGTGGAGAAGAGCTTCGAGACCGACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCAGAATCCTGCTGCTGAAGGTGGCCGGATTCAACCTGCTGATGACCCTGCGGCTGTGGAGCAGCGGCTCCCGGGCCAAGAGAAGCGGATCCGGCGCCACCAACTTCAGCCTGCTGAAGCAGGCCGGAGACGTGGAAGAAAACCCTGGCCCTAGGATGAGCATCGGCCTGCTGTGCTGCGCCGCCCTGAGCCTGCTGTGGGCAGGACCCGTGAACGCCGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAAACCGGCCAGAGCATGACCCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTATCGGCAGGACCCCGGCATGGGCCTGCGGCTGATCCACTACTCTGTGGGAGCCGGAATCACCGACCAGGGCGAGGTGCCCAACGGCTACAATGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCTGCCCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCTATGTGGGCAACACCGGCGAGCTGTTCTTCGGCGAGGGCTCCAGGCTGACCGTGCTGGAGGACCTGAAGAACGTGTTCCCCCCCGAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACACTGGTGTGTCTGGCCACCGGCTTCTACCCCGACCACGTGGAGCTGTCCTGGTGGGTGAACGGCAAGGAGGTGCACAGCGGCGTGTCTACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGACAGCCGGTACTGCCTGTCCTCCAGACTGAGAGTGAGCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCCGGTGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATTGTGAGCGCCGAGGCCTGGGGCAGGGCCGACTGCGGCTTCACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCTGCCCTGGTGCTGATGGCTATGGTGAAGCGGAAGGACAGCCGGGGCTAAGTCGAC
配列番号8 構成的−IL−7_T2A_TCR1のヌクレオチド配列。この配列は、NheI制限部位とSalI制限部位の間の領域を網羅する。
MFHVSFRYIFGLPPLILVLLPVASSDCDIEGKDGKQYESVLMVSIDQLLDSMKEIGSNCLNNEFNFFKRHICDANKEGMFLFRAARKLRQFLKMNSTGDFDLHLLKVSEGTTILLNCTGQVKGRKPAALGEAQPTKSLEENKSLKEQKKLNDLCFLKRLLQEIKTCWNKILMGTKEHGSRAKRSGSGEGRGSLLTCGDVEENPGPMETLLGLLILWLQLQWVSSKQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLTSLLLIQSSQREQTSGRLNASLDKSSGRSTLYIAASQPGDSATYLCAVRPLYGGSYIPTFGRGTSLIVHPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSSGSRAKRSGSGATNFSLLKQAGDVEENPGPRMSIGLLCCAALSLLWAGPVNAGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFFGEGSRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG
配列番号9 構成的IL−7_TCR1 IL7のタンパク質配列が下線で示され、TCRがイタリック体で示されている。
ACGCGTTAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTCTCGAGTCAGTGCTCTTTGGTGCCCATCAGGATCTTGTTCCAGCAGGTCTTGATTTCCTGCAGCAGCCGCTTCAGGAAGCACAGGTCGTTCAGTTTCTTCTGCTCTTTCAGGGACTTGTTCTCTTCCAGGCTCTTGGTAGGCTGGGCTTCTCCCAGGGCGGCAGGCTTTCTGCCCTTCACTTGGCCGGTGCAATTCAGCAGGATGGTGGTGCCCTCGGACACTTTCAGCAGATGCAGGTCGAAGTCGCCGGTGCTGTTCATCTTCAGGAACTGCCGCAGCTTTCTGGCGGCTCTGAACAGGAACATGCCTTCTTTGTTGGCGTCGCAGATGTGCCGCTTGAAGAAGTTGAACTCGTTGTTCAGGCAGTTGCTGCCGATTTCCTTCATGCTGTCCAGCAGCTGGTCGATGGACACCATCAGCACGCTCTCGTACTGCTTGCCGTCCTTGCCCTCGATGTCGCAGTCGCTGCTGGCCACAGGCAGCAGCACCAGGATCAGGGGGGGCAGGCCGAAGATGTACCGGAAGGACACGTGGAACATGGTGGCGGCCATATGGATCCAACGAATGTCGAGAGGCTGGATCGGTCCCGGTGTCTTCTATGGAGGTCAAAACAGCGTGGATGGCGTCTCCAGGCGATCTGACGGTTCACTAAACGAGCTCTGCTTATATAGGCCTCCCACCGTACACGCCTAGACGTCACGCCTTCTGTATGAAACAGTTTTTCCTCCACGCCTTCTGTATGAAACAGTTTTTCCTCCACGCCTTCTGTATGAAACAGTTTTTCCTCCGCTAGCGAGCTCAGGTACCGGCCAGTTAGGCCGATATCATGCATCGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCTGCAGTACGTGATTCTTGATCCCGAGCTTCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGCTTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCCTGGGCGCTGGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGATGACCTGCTGCGACGCTTTTTTTCTGGCAAGATAGTCTTGTAAATGCGGGCCAAGATCTGCACACTGGTATTTCGGTTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTGGCCGGCCTGCTCTGGTGCCTGGCCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTGCAGGGAGCTCAAAATGGAGGACGCGGCGCTCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAGGAAAAGGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACTGAGTACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGTGCTTTTGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTGGGTGGAGACTGAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCTTTTTGAGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAGGTGTCGTGAGCTAGCCGCCACCATGAAGAAGCACCTGACCACCTTTCTCGTGATCCTGTGGCTGTACTTCTACCGGGGCAACGGCAAGAACCAGGTGGAACAGAGCCCCCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCCCCCTTCAGCAACCTGCGGTGGTACAAGCAGGACACCGGCAGAGGCCCTGTGTCCCTGACCATCCTGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTACACCGCCACCCTGGACGCCGATACAAAGCAGAGCAGCCTGCACATCACCGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGGCGGCACAGACAGCTGGGGCAAGCTGCAGTTTGGCGCCGGAACACAGGTGGTCGTGACCCCCGACATCCAGAACCCTGACCCTGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTCAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCAGAATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCTGAGACTGTGGTCCAGCGGCAGCCGGGCCAAGAGATCTGGATCCGGCGCTACCAACTTTAGCCTGCTGAAGCAGGCCGGGGACGTGGAAGAAAACCCTGGCCCTAGGATGGCCAGCCTGCTGTTCTTCTGCGGCGCCTTCTACCTGCTGGGCACCGGCTCTATGGATGCCGACGTGACCCAGACCCCCCGGAACAGAATCACCAAGACCGGCAAGCGGATCATGCTGGAATGCTCCCAGACCAAGGGCCACGACCGGATGTACTGGTACAGACAGGACCCTGGCCTGGGCCTGCGGCTGATCTACTACAGCTTCGACGTGAAGGACATCAACAAGGGCGAGATCAGCGACGGCTACAGCGTGTCCAGACAGGCTCAGGCCAAGTTCAGCCTGTCCCTGGAAAGCGCCATCCCCAACCAGACCGCCCTGTACTTTTGTGCCACAAGCGGCCAGGGCGCCTACGAGGAGCAGTTCTTTGGCCCTGGCACCCGGCTGACAGTGCTGGAAGATCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTTCTGAGGCCGAAATCAGCCACACCCAGAAAGCCACACTCGTGTGTCTGGCCACCGGCTTCTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTGTCCACCGATCCCCAGCCTCTGAAAGAACAGCCCGCCCTGAACGACAGCCGGTACTGCCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCAGAAACCACTTCAGATGCCAGGTGCAGTTTTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACAGAGCCAAGCCCGTGACACAGATCGTGTCTGCCGAAGCTTGGGGGCGCGCCGATTGTGGCTTTACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGAAAGGCCACACTGTACGCCGTGCTGGTGTCTGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGCTAATAAGTCGAC
配列番号10 MluI制限部位とSalI制限部位の間の領域に及ぶセンス鎖5’から3’配列(図6参照)。
MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGKLQFGAGTQVVVTPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSSGSRAKRSGSGATNFSLLKQAGDVEENPGPRMASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFFGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG
配列番号11 TCR2配列
GCTAGCGCCGCCACCATGTTCCACGTGTCCTTCCGGTACATCTTCGGCCTGCCCCCCCTGATCCTGGTGCTGCTGCCTGTGGCCAGCAGCGACTGCGACATCGAGGGCAAGGACGGCAAGCAGTACGAGAGCGTGCTGATGGTGTCCATCGACCAGCTGCTGGACAGCATGAAGGAAATCGGCAGCAACTGCCTGAACAACGAGTTCAACTTCTTCAAGCGGCACATCTGCGACGCCAACAAAGAAGGCATGTTCCTGTTCAGAGCCGCCAGAAAGCTGCGGCAGTTCCTGAAGATGAACAGCACCGGCGACTTCGACCTGCATCTGCTGAAAGTGTCCGAGGGCACCACCATCCTGCTGAATTGCACCGGCCAAGTGAAGGGCAGAAAGCCTGCCGCCCTGGGAGAAGCCCAGCCTACCAAGAGCCTGGAAGAGAACAAGTCCCTGAAAGAGCAGAAGAAACTGAACGACCTGTGCTTCCTGAAGCGGCTGCTGCAGGAAATCAAGACCTGCTGGAACAAGATCCTGATGGGCACCAAAGAGCACGGAAGCAGAGCCAAGAGAAGCGGCTCTGGCGCGCCCGTGAAGCAGACCCTGAACTTCGACCTGCTGAAACTGGCCGGCGACGTGGAAAGCAACCCTGGCCCTATGAAGAAGCACCTGACCACCTTTCTCGTGATCCTGTGGCTGTACTTCTACCGGGGCAACGGCAAGAACCAGGTGGAACAGAGCCCCCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACTCTGCAGTGCAACTACACCGTGTCCCCCTTCAGCAACCTGCGCTGGTACAAGCAGGATACCGGCAGAGGCCCTGTGTCCCTGACCATCCTGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTACACCGCCACCCTGGACGCCGATACAAAGCAGAGCAGCCTGCACATCACCGCCTCCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGGCGGCACAGACAGCTGGGGCAAGCTGCAGTTTGGCGCCGGAACACAGGTGGTCGTGACCCCCGACATCCAGAACCCTGACCCTGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACTCCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGACCGTGCTGGATATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCTAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAACTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAATTTCCAGAACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGATTCAACCTGCTGATGACCCTGCGGCTGTGGTCCTCTGGCTCTCGGGCCAAGAGAAGCGGCAGCGGCGCCACCAATTTCAGCCTGCTGAAGCAGGCAGGGGATGTGGAAGAGAATCCCGGCCCTAGAATGGCCTCCCTGCTGTTTTTCTGCGGCGCCTTCTACCTGCTGGGGACCGGCAGCATGGACGCTGACGTGACCCAGACCCCCCGGAACAGAATCACCAAGACCGGCAAGCGGATCATGCTGGAATGCAGCCAGACAAAGGGCCACGACCGGATGTACTGGTACAGACAGGATCCAGGACTGGGCCTGAGGCTGATCTACTACAGCTTCGATGTGAAGGACATCAACAAGGGCGAGATCAGCGACGGCTACAGCGTGTCCAGACAGGCCCAGGCCAAGTTCTCCCTGAGCCTGGAAAGCGCCATCCCCAACCAGACCGCCCTGTACTTTTGTGCCACAAGCGGCCAGGGCGCCTACGAGGAACAGTTCTTTGGCCCTGGCACCCGGCTGACAGTGCTGGAAGATCTGAAGAACGTGTTCCCCCCAGAGGTGGCAGTGTTCGAGCCTAGCGAGGCCGAGATCTCCCACACCCAGAAAGCCACACTCGTGTGTCTGGCCACCGGATTCTACCCCGACCATGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTGTCCACCGATCCCCAGCCTCTGAAAGAACAGCCCGCCCTGAACGACAGCCGGTACTGCCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCAGAAATCACTTCAGATGCCAGGTGCAGTTTTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGATAGGGCCAAGCCCGTGACTCAGATCGTGTCTGCCGAAGCCTGGGGCAGAGCCGATTGCGGCTTTACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACACTGTACGCCGTGCTGGTGTCTGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGCTGATGAGGTCGAC
配列番号12 NheI制限部位とSalI制限部位の間の領域を網羅するヌクレオチド配列。
MFHVSFRYIFGLPPLILVLLPVASSDCDIEGKDGKQYESVLMVSIDQLLDSMKEIGSNCLNNEFNFFKRHICDANKEGMFLFRAARKLRQFLKMNSTGDFDLHLLKVSEGTTILLNCTGQVKGRKPAALGEAQPTKSLEENKSLKEQKKLNDLCFLKRLLQEIKTCWNKILMGTKEHGSRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGKLQFGAGTQVVVTPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSSGSRAKRSGSGATNFSLLKQAGDVEENPGPRMASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFFGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG**
配列番号13 構成的IL−7_TCR2の翻訳されたタンパク質配列
GGAGGAAAAACTGTTTCATACAGAAGGCGT
配列番号14 ヒトIL2プロモーターNFAT TRE
TAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCAGCCTCTCGACATTCGTTGGATC
配列番号15 最小CMVプロモーター
Claims (49)
- 核酸構築物であって、
(i)生理活性分子をコードする第1のヌクレオチド配列と、
(ii)抗原受容体をコードする第2のヌクレオチド配列と、
(iii)前記第1のヌクレオチド配列に作動可能に連結された誘導性プロモーターと、
(iv)前記第2のヌクレオチドに作動可能に連結された構成的プロモーターと、を含む、核酸構築物。 - 前記誘導性プロモーターからの発現は、T細胞の活性化によって誘導される、請求項1に記載の核酸構築物。
- 前記誘導性プロモーターは、活性化T細胞核内因子(NFAT)転写応答エレメント(TRE)を含む、請求項1または2のいずれか一項に記載の核酸構築物。
- 前記NFAT TREは、配列番号14の核酸配列またはその変異体を有する、請求項1〜3のいずれか一項に記載の核酸構築物。
- 前記誘導性プロモーターは、前記NFAT TREの3つ以上のコピーを含む、請求項1〜4のいずれか一項に記載の核酸構築物。
- 前記核酸構築物は、配列番号1の配列を含む、請求項1〜5のいずれか一項に記載の核酸構築物。
- 前記構成的プロモーターは、ヒト伸長因子1αプロモーターである、請求項1〜6のいずれか一項に記載の核酸構築物。
- 前記第1のヌクレオチド配列はサイトカインをコードする、請求項1〜7のいずれか一項に記載の核酸構築物。
- 前記サイトカインはインターロイキン7(IL−7)である、請求項8に記載の核酸構築物。
- 前記IL−7はヒトIL−7である、請求項9に記載の核酸構築物。
- 前記抗原受容体はT細胞受容体(TCR)である、請求項1〜10のいずれか一項に記載の核酸構築物。
- 前記TCRは高親和性TCRである、請求項11に記載の核酸構築物。
- 前記TCRは、抗原のペプチド断片を提示するMHCに特異的に結合する、請求項11または請求項12に記載の核酸構築物。
- 前記抗原は、癌細胞によって発現される腫瘍抗原である、請求項13に記載の核酸構築物。
- 前記TCRは、配列番号5、6または11のいずれか1つのアミノ酸配列を含む、請求項11〜14のいずれか一項に記載の核酸構築物。
- 前記抗原受容体はキメラ抗原受容体(CAR)である、請求項1〜10のいずれか一項に記載の核酸構築物。
- 前記CARは、細胞によって発現される抗原に特異的に結合する、請求項16に記載の核酸構築物。
- 前記抗原は、前記癌細胞によって発現される腫瘍抗原である、請求項17に記載の核酸構築物。
- 前記腫瘍抗原はNY−ESO−1またはMAGE−A10である、請求項14または18に記載の核酸構築物。
- 前記第1のヌクレオチド配列は、第1の方向に発現するように構成され、前記第2のヌクレオチド配列は、第1の方向に発現するように構成される、請求項1〜19のいずれか一項に記載の核酸構築物。
- 請求項1〜20のいずれか一項に記載の核構築物を含む、ベクター。
- 前記ベクターはレンチウイルスベクターである、請求項21に記載のベクター。
- 請求項21または22に記載のベクターを含む、ウイルス粒子。
- 請求項1〜23のいずれか一項に記載の核構築物、ベクター、またはウイルス粒子を含む、T細胞集団。
- T細胞活性化に伴って、異種抗原受容体を構成的に発現し、生理活性分子を誘導性に発現する、T細胞集団。
- 前記生理活性分子をコードする前記ヌクレオチド配列は、誘導性プロモーターに作動可能に連結され、前記異種抗原受容体をコードする前記ヌクレオチド配列は、構成的プロモーターに作動可能に連結される、請求項25に記載の集団。
- 前記誘導性プロモーターは、活性化T細胞核内因子(NFAT)転写応答エレメント(TRE)を含む、請求項26に記載の集団。
- 前記NFAT TREは、配列番号14の核酸配列またはその変異体を有する、請求項27に記載の集団。
- 前記誘導性プロモーターは、前記NFAT TREの3つ以上のコピーを含む、請求項26〜28のいずれか一項に記載の集団。
- 前記構成的プロモーターは、ヒト伸長因子1αプロモーターである、請求項26〜29のいずれか一項に記載の集団。
- 前記第1のヌクレオチド配列はサイトカインをコードする、請求項25〜30のいずれか一項に記載の集団。
- 前記サイトカインはインターロイキン7(IL−7)である、請求項31に記載の集団。
- 前記抗原受容体は、請求項11〜17のいずれか一項に定義される通りである、請求項25〜32のいずれか一項に記載の集団。
- 請求項24〜33のいずれか一項に記載のT細胞集団および薬学的に許容される賦形剤を含む、医薬組成物。
- ヒトまたは動物の体の治療方法に使用するための、請求項24〜33のいずれか一項に記載のT細胞集団。
- 個体における癌の治療方法に使用するための、請求項24〜33のいずれか一項に記載のT細胞集団。
- 個体における癌の治療のための薬物の製造における、請求項24〜33のいずれか一項に記載のT細胞集団の使用。
- 癌を有する個体に請求項24〜33のいずれか一項に記載のT細胞集団を投与することを含む、癌を治療する方法。
- 請求項36による使用のための集団、請求項37による使用、または請求項38による方法であって、前記癌は、前記抗原受容体に結合する1つ以上の癌細胞の存在によって特徴付けられる、使用のための集団、使用、または方法。
- 前記T細胞は自己由来である、請求項36〜39のいずれか一項に記載の、使用のための集団、使用、または方法。
- 前記T細胞は同種由来である、請求項36〜40のいずれか一項に記載の、使用のための集団、使用、または方法。
- 改変T細胞集団を生成する方法であって、
請求項1〜22のいずれか一項に記載の核酸構築物またはベクターを、ドナー個体から得られたT細胞集団に導入して改変T細胞集団を生成すること、を含む、方法。 - 前記改変T細胞を拡大培養するおよび/または保存する、請求項42に記載の方法。
- 前記改変T細胞集団を薬学的に許容される賦形剤と配合することを含む、請求項42または43に記載の方法。
- 前記改変T細胞集団をレシピエント個体に投与することを含む、請求項42〜44のいずれか一項に記載の方法。
- 前記レシピエント個体は癌状態を有する、請求項45に記載の方法。
- 前記癌状態は、前記抗原受容体に結合する1つ以上の癌細胞の存在によって特徴付けられる、請求項46に記載の方法。
- 前記ドナー個体と前記レシピエント個体とが同じである、請求項42〜47のいずれか一項に記載の方法。
- 前記ドナー個体と前記レシピエント個体とが異なる、請求項42〜47のいずれか一項に記載の方法。
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GBGB1713078.2A GB201713078D0 (en) | 2017-08-15 | 2017-08-15 | T Cell Modification |
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PCT/EP2018/072148 WO2019034703A2 (en) | 2017-08-15 | 2018-08-15 | MODIFICATION OF LYMPHOCYTES T |
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Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB201713078D0 (en) * | 2017-08-15 | 2017-09-27 | Adaptimmune Ltd | T Cell Modification |
AU2019253562A1 (en) * | 2018-04-09 | 2020-11-19 | The Trustees Of The University Of Pennsylvania | Methods and compositions comprising a viral vector for expression of a transgene and an effector |
GB201819540D0 (en) * | 2018-11-30 | 2019-01-16 | Adaptimmune Ltd | T cell modification |
EP3753566A1 (en) * | 2019-06-21 | 2020-12-23 | Medizinische Hochschule Hannover | All-in one viral vector for car and therapeutic effector molecule |
CN114867490A (zh) * | 2019-09-20 | 2022-08-05 | 克莱格医学有限公司 | 细胞因子调控表达的免疫效应细胞 |
WO2021259334A1 (zh) * | 2020-06-24 | 2021-12-30 | 南京博望医药科技有限公司 | 自我调节型嵌合抗原受体及其在肿瘤免疫中的应用 |
WO2022057941A1 (zh) * | 2020-09-21 | 2022-03-24 | 克莱格医学有限公司 | 表达il-7r结合蛋白的免疫效应细胞 |
TW202340474A (zh) * | 2022-01-11 | 2023-10-16 | 大陸商深圳市珈鈺生物科技有限公司 | 樹突細胞腫瘤疫苗和其用途 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH10179181A (ja) * | 1989-01-23 | 1998-07-07 | Chiron Corp | 感染および過剰増殖障害の為の組換え療法 |
US20120071859A1 (en) * | 2009-04-30 | 2012-03-22 | Morgan Richard A | Inducible interleukin-12 |
WO2016073755A2 (en) * | 2014-11-05 | 2016-05-12 | Board Of Regents, The University Of Texas System | Gene modified immune effector cells and engineered cells for expansion of immune effector cells |
WO2016126608A1 (en) * | 2015-02-02 | 2016-08-11 | Novartis Ag | Car-expressing cells against multiple tumor antigens and uses thereof |
Family Cites Families (36)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6235525B1 (en) | 1991-05-23 | 2001-05-22 | Ludwig Institute For Cancer Research | Isolated nucleic acid molecules coding for tumor rejection antigen precursor MAGE-3 and uses thereof |
US5342774A (en) | 1991-05-23 | 1994-08-30 | Ludwig Institute For Cancer Research | Nucleotide sequence encoding the tumor rejection antigen precursor, MAGE-1 |
US5541104A (en) | 1991-05-23 | 1996-07-30 | Ludwig Institute For Cancer Research | Monoclonal antibodies which bind to tumor rejection antigen precursor mage-1 |
JP3608788B2 (ja) | 1992-08-31 | 2005-01-12 | ルドヴィグ・インスティテュート・フォー・キャンサー・リサーチ | Mage−3遺伝子から誘導されてhla−a1により提示される単離されたノナペプチドおよびそれらの用途 |
US6222012B1 (en) | 1992-08-31 | 2001-04-24 | Ludwig Institute For Cancer Research | Isolated nonapeptides presented by HLA molecules, and uses thereof |
DE69524264T2 (de) | 1994-03-01 | 2002-07-18 | Ludwig Inst Cancer Res | Bestimmung krebsartiger beschaffenheiten durch die genexpression eines mitgliedes der melanomartigengruppe, mage |
US6291430B1 (en) | 1997-09-12 | 2001-09-18 | Ludwig Institute For Cancer Research | Mage-3 peptides presented by HLA class II molecules |
EP1143013A1 (en) * | 2000-04-03 | 2001-10-10 | Warner-Lambert Company | Methods and compositions for screening Icrac modulators |
WO2013126726A1 (en) * | 2012-02-22 | 2013-08-29 | The Trustees Of The University Of Pennsylvania | Double transgenic t cells comprising a car and a tcr and their methods of use |
WO2017133175A1 (en) * | 2016-02-04 | 2017-08-10 | Nanjing Legend Biotech Co., Ltd. | Engineered mammalian cells for cancer therapy |
CN108884459B (zh) * | 2016-04-26 | 2024-04-02 | 科济生物医药(上海)有限公司 | 一种改善免疫应答细胞功能的方法 |
GB201608052D0 (en) * | 2016-05-09 | 2016-06-22 | Univ Oslo Hf | T-cell receptors which recognise frameshift mutants of TGF-beta RII |
CN106086073A (zh) * | 2016-07-01 | 2016-11-09 | 西北民族大学 | 一种细胞因子il‑7的表达方法 |
WO2018050225A1 (en) * | 2016-09-15 | 2018-03-22 | Yu Di | T-cell immunotherapy |
GB201616238D0 (en) * | 2016-09-23 | 2016-11-09 | Adaptimmune Ltd | Modified T cells |
US20200010803A1 (en) * | 2017-03-08 | 2020-01-09 | Memorial Sloan Kettering Cancer Center | Immune cell compositions and methods of use |
US20200255505A1 (en) * | 2017-08-09 | 2020-08-13 | Eureka Therapeutics, Inc. | Cells Expressing Cell Surface Receptors and Antibodies |
GB201713078D0 (en) * | 2017-08-15 | 2017-09-27 | Adaptimmune Ltd | T Cell Modification |
AU2019253562A1 (en) * | 2018-04-09 | 2020-11-19 | The Trustees Of The University Of Pennsylvania | Methods and compositions comprising a viral vector for expression of a transgene and an effector |
WO2020028656A1 (en) * | 2018-08-01 | 2020-02-06 | Nantkwest, Inc. | A quadricistronic system comprising a homing receptor or a cytokine, and chimeric antigen receptor for genetic modification of immunotherapies |
CN116286665A (zh) * | 2018-11-16 | 2023-06-23 | 上海煦顼技术有限公司 | 嵌合抗原受体细胞分泌治疗剂 |
GB201819540D0 (en) * | 2018-11-30 | 2019-01-16 | Adaptimmune Ltd | T cell modification |
CA3132660A1 (en) * | 2019-03-06 | 2020-09-10 | Lentigen Technology, Inc. | Compositions and methods for treating cancer with self-driving chimeric antigen receptors |
RU2742000C2 (ru) * | 2019-03-13 | 2021-02-01 | Общество С Ограниченной Ответственностью "Анабион" | Выделенный альтернативный внутриклеточный сигнальный домен химерного антигенного рецептора и включающий его химерный антигенный рецептор |
US11788072B2 (en) * | 2019-04-30 | 2023-10-17 | Innovative Cellular Therapeutics Holdings, Ltd. | Activation of APC in immunotherapy |
WO2021028359A1 (en) * | 2019-08-09 | 2021-02-18 | Sangamo Therapeutics France | Controlled expression of chimeric antigen receptors in t cells |
CN112980886B (zh) * | 2019-12-02 | 2022-02-22 | 河北森朗生物科技有限公司 | 一种能高效制备且应用安全的嵌合抗原受体t细胞及其制备方法与应用 |
US20210252059A1 (en) * | 2020-02-19 | 2021-08-19 | Innovative Cellular Therapeutics Holdings, Ltd. | Engineered safety in cell therapy |
CN111575297A (zh) * | 2020-05-29 | 2020-08-25 | 上海斯丹赛生物技术有限公司 | 诱导型Fms相关酪氨酸激酶3配体修饰的细胞及其用途 |
WO2021244654A1 (en) * | 2020-06-05 | 2021-12-09 | Nanjing Legend Biotech Co., Ltd. | Activation induced cytokine production in immune cells |
WO2021259334A1 (zh) * | 2020-06-24 | 2021-12-30 | 南京博望医药科技有限公司 | 自我调节型嵌合抗原受体及其在肿瘤免疫中的应用 |
CN112175998A (zh) * | 2020-10-12 | 2021-01-05 | 广东昭泰体内生物医药科技有限公司 | 一种嵌合抗原受体t细胞及其应用 |
CN112226462A (zh) * | 2020-10-12 | 2021-01-15 | 广东昭泰体内生物医药科技有限公司 | 共表达分泌型il-7和选择性ccl19的表达载体及其应用 |
CN112210540A (zh) * | 2020-10-12 | 2021-01-12 | 广东昭泰体内生物医药科技有限公司 | 一种表达免疫调控因子的car-t细胞及其应用 |
CN112210539A (zh) * | 2020-10-12 | 2021-01-12 | 广东昭泰体内生物医药科技有限公司 | 一种第四代car-t细胞及其应用 |
WO2023114777A2 (en) * | 2021-12-14 | 2023-06-22 | The Trustees Of The University Of Pennsylvania | Cd5 modified cells comprising chimeric antigen receptors (cars) for treatment of solid tumors |
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2017
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH10179181A (ja) * | 1989-01-23 | 1998-07-07 | Chiron Corp | 感染および過剰増殖障害の為の組換え療法 |
US20120071859A1 (en) * | 2009-04-30 | 2012-03-22 | Morgan Richard A | Inducible interleukin-12 |
WO2016073755A2 (en) * | 2014-11-05 | 2016-05-12 | Board Of Regents, The University Of Texas System | Gene modified immune effector cells and engineered cells for expansion of immune effector cells |
WO2016126608A1 (en) * | 2015-02-02 | 2016-08-11 | Novartis Ag | Car-expressing cells against multiple tumor antigens and uses thereof |
Non-Patent Citations (3)
Title |
---|
KOUSKOFF, V. ET AL.: ""Cassette vectors directing expression of T cell receptor genes in transgenic mice"", J. IMMUNOL. METHODS, vol. 180, JPN6022025650, pages 273 - 280, XP004021050, ISSN: 0004983002, DOI: 10.1016/0022-1759(95)00002-R * |
OVERWIJK, W. W. ET AL.: ""Tumor regression and autoimmunity after reversal of a functionally tolerant state of self-reactive", J. EXP. MED., vol. 198, JPN6022025648, 2003, pages 569 - 580, XP002416108, ISSN: 0004983001, DOI: 10.1084/jem.20030590 * |
ZHANG, L. ET AL.: ""Improving adoptive T cell therapy by targeting and controlling IL-12 expression to the tumor enviro", MOL. THER., vol. 19, JPN6022025649, 2011, pages 751 - 759, XP055327746, ISSN: 0004983000, DOI: 10.1038/mt.2010.313 * |
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MX2020001791A (es) | 2020-07-29 |
US11111301B2 (en) | 2021-09-07 |
CN111315402B (zh) | 2024-05-07 |
CA3072841A1 (en) | 2019-02-21 |
RU2020110331A3 (ja) | 2022-04-15 |
BR112020003007A2 (pt) | 2020-08-11 |
KR20200039765A (ko) | 2020-04-16 |
US20190100592A1 (en) | 2019-04-04 |
RU2020110331A (ru) | 2021-09-16 |
WO2019034703A3 (en) | 2019-03-28 |
CN111315402A (zh) | 2020-06-19 |
WO2019034703A2 (en) | 2019-02-21 |
GB201713078D0 (en) | 2017-09-27 |
US20220064305A1 (en) | 2022-03-03 |
EP3668537A2 (en) | 2020-06-24 |
JP7289293B2 (ja) | 2023-06-09 |
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