CN108779174B - 靶向her2的嵌合抗原受体 - Google Patents

靶向her2的嵌合抗原受体 Download PDF

Info

Publication number
CN108779174B
CN108779174B CN201680076412.0A CN201680076412A CN108779174B CN 108779174 B CN108779174 B CN 108779174B CN 201680076412 A CN201680076412 A CN 201680076412A CN 108779174 B CN108779174 B CN 108779174B
Authority
CN
China
Prior art keywords
gly
leu
ser
pro
thr
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201680076412.0A
Other languages
English (en)
Other versions
CN108779174A (zh
Inventor
S.J.普赖斯曼
S.J.福曼
C.E.布朗
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
City of Hope
Original Assignee
City of Hope
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by City of Hope filed Critical City of Hope
Priority to CN202211409883.2A priority Critical patent/CN116063574A/zh
Publication of CN108779174A publication Critical patent/CN108779174A/zh
Application granted granted Critical
Publication of CN108779174B publication Critical patent/CN108779174B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001102—Receptors, cell surface antigens or cell surface determinants
    • A61K39/001103—Receptors for growth factors
    • A61K39/001106—Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ErbB4
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/461—Cellular immunotherapy characterised by the cell type used
    • A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/463—Cellular immunotherapy characterised by recombinant expression
    • A61K39/4631—Chimeric Antigen Receptors [CAR]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
    • A61K39/4643—Vertebrate antigens
    • A61K39/4644—Cancer antigens
    • A61K39/464402—Receptors, cell surface antigens or cell surface determinants
    • A61K39/464403—Receptors for growth factors
    • A61K39/464406—Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ ErbB4
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70514—CD4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70517—CD8
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70521—CD28, CD152
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70578—NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/71—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/515—Animal cells
    • A61K2039/5156—Animal cells expressing foreign proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/515—Animal cells
    • A61K2039/5158—Antigen-pulsed cells, e.g. T-cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/80—Vaccine for a specifically defined cancer
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/80—Vaccine for a specifically defined cancer
    • A61K2039/812—Breast
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
    • A61K2239/26—Universal/off- the- shelf cellular immunotherapy; Allogenic cells or means to avoid rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/524—CH2 domain
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/71—Decreased effector function due to an Fc-modification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/33—Fusion polypeptide fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16041—Use of virus, viral particle or viral elements as a vector
    • C12N2740/16043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2830/00—Vector systems having a special element relevant for transcription
    • C12N2830/15—Vector systems having a special element relevant for transcription chimeric enhancer/promoter combination

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • General Engineering & Computer Science (AREA)
  • Oncology (AREA)
  • Hematology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Virology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Developmental Biology & Embryology (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

描述了嵌合跨膜免疫受体(CAR)，其包含靶向HER2的胞外域、跨膜区、共刺激域和胞内信号传导域。

Description

靶向HER2的嵌合抗原受体
发明背景
已经研究了基于肿瘤特异性T细胞的免疫疗法，包括采用工程化T细胞的疗法的抗肿瘤治疗。在某些情况下，用于此类疗法的T细胞在体内未持续足够长的时段保持有活性。在某些情况下，部分由于实体瘤的异质性性质和在靶向自身抗原时对非癌性细胞的脱靶效应的潜力，T细胞的肿瘤特异性是相对较低的。因此，本领域需要具有改善的抗肿瘤特异性和功能的肿瘤特异性癌症疗法。
嵌合抗原受体(CAR)由胞外肿瘤识别/靶向域、胞外接头/间隔物、跨膜域和胞内T细胞活化和共刺激信号传导域构成。识别/目标区域的设计对于避免有害的脱靶效应至关重要。大多数CAR肿瘤靶向域是源自抗体序列的单链可变片段(scFv)，其利用抗体与特定抗原结合的特异性。也有来自正常受体配体的CAR肿瘤靶向域的实例，例如靶向表达IL-13受体IL-13Rα2的细胞的IL-13细胞因子CAR。
利用表达CAR的工程化T细胞的过继T细胞疗法(ACT)已经在具有CD19+B细胞恶性的患者中表明强力且持久的临床效应(Priceman et al.2015 Curr Opin Oncol；Maus etal.2014Blood 123:2625-2635)。随着血液学疾病中的早期成功，此办法对实体瘤的更宽的应用现在在密集的研究下。
根据国家癌症研究院(National Cancer Institute)监测、流行病学和最终结果项目(Surveillance,Epidemiology,and End Results Program,SEER)数据，2014年估计有40,000人死于乳腺癌，主要来自转移性疾病。约25-30％的乳腺癌患者携带HER2基因的扩增，其赋予特别差的预后。即使随着包括靶向疗法在内的较新型试剂的出现，IV期疾病的总体死亡率也仅有适度的改善。例如，在HER2阳性乳腺癌患者的随机化试验中，两种HER2靶向抗体曲妥珠单抗(trastuzumab)和帕妥珠单抗(pertuzumab)加多西他赛(docetaxel)的最有希望的治疗组合相对于单独的曲妥珠单抗和多西他赛产生56.5个月的中值总存活和仅6.3个月的无进展存活的延长。
发明概述
本文描述了嵌合跨膜免疫受体(嵌合抗原受体或“CAR”)，其包含胞外域、跨膜区和胞内信号传导域。胞外域包括靶向HER2的scFv和任选地间隔物，所述间隔物包含例如人Fe域的一部分。跨膜部分包括例如CD4跨膜域、CD8跨膜域、CD28跨膜域或CD3跨膜域。胞内信号传导域包括来自人CD3复合物的ζ链(CD3ξ)的信号传导域和一个或多个共刺激域，例如4-1BB或CD28共刺激域。胞外域使CAR在T细胞表面上表达时能够将T细胞活性导向表达HER2的那些细胞。此类细胞包括某些乳腺癌细胞和某些脑癌细胞。在胞内区域中包含与CD3ζ串联的共刺激域，如4-1BB(CD137)共刺激域使得T细胞能够接收共刺激信号。可以将T细胞，例如患者特异性自体T细胞工程化改造为表达本文所述的CAR，并且可以将工程化细胞扩充并用于ACT。可以使用各种T细胞亚组，包括αβT细胞和γδT细胞两者。另外，可以在其他免疫细胞如NK细胞中表达CAR。在用表达本文所述的CAR的免疫细胞治疗患者的情况下，细胞可以是自体T细胞或同种异体T细胞。在一些情况下，使用的细胞是包括CD62L+、CCR7+、CD45RO+和CD45RA-的CD4+和CD8+中央记忆T细胞(TCM)的细胞群体，或者使用的细胞是包括CD4+和CD8+TCM细胞、干中央记忆T细胞和幼稚T细胞的细胞群体(即TCM/SCM/N细胞的群体)。TCM/SCM/N细胞群体是CD62L+，CCR7+并且包括CD45RA+和CD45RO+细胞两者以及CD4+细胞和CD8+细胞两者。与使用其他类型的患者特异性T细胞相比，使用此类细胞可以改善过继转移后细胞的长期持久性。
本文中描述了编码CAR的核酸分子，所述CAR包含：靶向HER2的scFv(例如DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKGSTSGGGSGGGSGGGGSSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS；SEQ ID NO:1)或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；选自以下的跨膜域：CD4跨膜域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体、CD8跨膜域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体、CD28跨膜域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体、和CD3ξ跨膜域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；共刺激域(例如CD28共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；或4-I BB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；或CD28共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体和4-1BB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体两者；和CD3ξ信号传导域或其具有1-5个(例如1或2个)氨基酸修饰的变体。
在各个实施方案中：共刺激域选自下组：CD28共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体、4-1BB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体和OX40共刺激域或其具有1-5个(例如1或2个)氨基酸修饰的变体。在某些实施方案中，存在4-1BB共刺激域或或其具有1-5个(例如1或2个)氨基酸修饰的变体。在一些实施方案中，存在有两个共刺激域，例如CD28共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体和4-1BB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体。在各个实施方案中，1-5个(例如1或2个)氨基酸修饰是取代。
在一些情况下，共刺激域和CD3ξ信号传导域和/或两个共刺激域之间有1-6个氨基酸(例如GGG)的短序列。
另外的实施方案，CAR包含：靶向HER2的scFv；选自下组的两种不同的共刺激域：CD28共刺激域或其具有1-5个(例如1或2个)氨基酸修饰的变体、4-lBB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰的变体和OX40共刺激域或其具有1-5个(例如1或2个)氨基酸修饰的变体；选自下组的两种不同的共刺激域：CD28共刺激域或其具有1-2个氨基酸修饰的变体、4-1BB共刺激域或其具有1-2个氨基酸修饰的变体和OX40共刺激域或其具有1-2个氨基酸修饰的变体；HER2 scFv或其具有1-2个氨基酸修饰的变体；选自以下的跨膜域：CD4跨膜域或其具有1-2个氨基酸修饰的变体、CD8跨膜域或其具有1-2个氨基酸修饰的变体、CD28跨膜域或其具有1-2个氨基酸修饰的变体、和CD3ξ跨膜域或其具有1-2个氨基酸修饰的变体；共刺激域(例如CD28共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；或4-1BB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；或CD28共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体和4-1BB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体两者；和CD3ξ信号传导域或其具有1-2个氨基酸修饰的变体；位于HER2 scFv或其变体和跨膜域之间的间隔物区(例如间隔物区包含选自下组的氨基酸序列：SEQ ID NO:2-12和42(表3)或其具有1-5个(例如1或2个)氨基酸修饰的变体)；间隔物包含IgG铰链区；间隔物区包含1-150个氨基酸；没有间隔物；4-1BB信号传导域包含SEQ ID NO:24的氨基酸序列，CD3ξ信号传导域包含SEQ ID NO:21的氨基酸序列和位于共刺激域和CD3ξ信号传导域或其变体之间的3至15个氨基酸的接头。在有两个共刺激域的某些实施方案中，一个是4-1BB共刺激域，而另一个是选自以下的共刺激域：CD28和CD28gg。在各个实施方案中，1-5个(例如1或2)氨基酸修饰是取代，例如保守取代。
在一些实施方案中：核酸分子表达包含选自SEQ ID NO:26-41的氨基酸序列的多肽；嵌合抗原受体包含选自SEQ ID NO:26-41的氨基酸序列。
还公开了通过载体转导的人T细胞群体，所述载体包含编码嵌合抗原受体的表达盒，其中嵌合抗原受体包含：靶向HER2的scFv；选自以下的跨膜域：CD4跨膜域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体、CD8跨膜域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体、CD28跨膜域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体、和CD3ξ跨膜域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；共刺激域(例如CD28共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；或4-1BB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；或CD28共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体和4-1BB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体两者；和CD3ξ信号传导域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体。在各个实施方案中：人T细胞群体包含表达嵌合抗原受体的载体，所述嵌合抗原受体包含选自以下的氨基酸序列：SEQ ID NO:26-41中任一项或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体；人T细胞群体包含中央记忆T细胞(TCM细胞)，例如至少20％、30％、40％、50％60％、70％、80％的细胞是TCM细胞，或T细胞群体包含中央记忆T细胞、幼稚T细胞和干中央记忆细胞的组合(TCM/SCM/N细胞)，例如至少20％、30％、40％、50％、60％、70％、80％的细胞是TCM/SCM/N细胞。在任一情况下，T细胞群体包含CD4+细胞和CD8+细胞两者(例如，至少20％的CD3+T细胞是CD4+，并且至少3％的CD3+T细胞是CD8+，并且至少70、80或90％是CD4+或CD8+；至少15％、20％、25％、30％、35％、40％、50％、60％的细胞CD3+细胞是CD4+，并且至少4％、5％、8％、10％、20的CD3+细胞是CD8+细胞)。
还描述了治疗患者中的癌症的方法，其包括施用通过包含编码嵌合抗原受体的表达盒的载体转导的自体或同种异体人T细胞群体(例如自体或同种异体T细胞，其包含中央记忆T细胞(TCM细胞)或中央记忆T细胞、幼稚T细胞和干中央记忆细胞的组合(即，T细胞是TCM/SCM/N细胞)。至少20％、30％、40％、50％60％、70％、80％的细胞是TCM/SCM/N细胞。在任一种情况下，T细胞群体包含CD4+细胞和CD8+细胞两者(例如，至少20％的CD3+T细胞是CD4+，并且至少3％的CD3+T细胞是CD8+，并且至少70、80或90％是CD4+或CD8+；至少15％、20％、25％、30％、35％、40％、50％、60％的细胞CD3+细胞是CD4+，并且至少4％、5％、8％、10％、20的CD3+细胞是CD8+细胞)，其中嵌合抗原受体包含选自以下的氨基酸序列：SEQ ID NO:26-41或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体。在各个实施方案中：癌症是脑癌，例如，作为从乳腺癌的转移的HER2表达性脑癌；并且通过方法制备经转导的人T细胞，所述方法包括从患者获得T细胞，处理T细胞以分离中央记忆T细胞，并且用包含编码嵌合抗原受体的表达盒的病毒载体转导中央记忆细胞的至少一部分，其中嵌合抗原受体包含选自下组的氨基酸序列：SEQ ID NO:26或27或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体。在一些情况下，不直接对脑肿瘤施用CAR T细胞，而是在患者的脑内对室内空间施用。
还描述了：编码多肽的核酸分子，所述多肽包含与选自SEQ ID NO 26-41的氨基酸序列至少95％相同的氨基酸序列；编码包含氨基酸序列的多肽的核酸分子，所述氨基酸序列除了存在不超过5个氨基酸取代、缺失或插入外与选自SEQ ID NO:26-41的氨基酸序列相同；编码包含氨基酸序列的多肽的核酸分子，所述氨基酸序列除了存在不超过5个氨基酸取代之外与选自SEQ ID NO:26-41的氨基酸序列相同；和编码包含氨基酸序列的多肽的核酸分子，所述氨基酸序列除了存在不超过2个氨基酸取代以外与选自SEQ ID NO:26-41的氨基酸序列相同。
本公开内容还包括编码本文所述的任何CAR的核酸分子(例如包含编码CAR之一的核酸序列的载体)和表达本文所述的任何CAR的分离的T淋巴细胞。
本文描述的CAR可以包括位于HER2结合域(例如，HER2 scFv)和跨膜域之间的间隔物区。可以使用多种不同的间隔物。它们中的一些包括人Fc区的至少一部分，例如人Fc区的铰链部分或CH3域或其变体。下面的表1提供了可用于本文所述的CAR中的各种间隔物。
表1：间隔物的实例
Figure GDA0003366579280000061
Figure GDA0003366579280000071
一些间隔物区包括整个或部分的免疫球蛋白(例如，IgG1、IgG2、IgG3、IgG4)铰链区，即落在免疫球蛋白的CH1和CH2域之间的序列，例如，IgG4 Fc铰链或CD8铰链。一些间隔物区包含免疫球蛋白CH3域或CH3域和CH2域两者。免疫球蛋白来源的序列可以包含一个或多个氨基酸修饰，例如，1、2、3、4或5个取代，例如，减少脱靶结合的取代。
“氨基酸修饰”指蛋白质或肽序列中的氨基酸取代、插入，和/或缺失。“氨基酸取代”或“取代”指亲本肽或蛋白质序列中特定位置上的氨基酸被另一氨基酸替换。可以进行取代用于以非保守方式(即，通过将密码子从属于具有特定大小或特征的氨基酸分组的氨基酸改变成属于另一分组的氨基酸)或以保守方式(即，通过将密码子从属于具有特定大小或特征的氨基酸分组的氨基酸改变成属于相同分组的氨基酸)来改变所得蛋白质中的氨基酸。该保守改变一般导致所得蛋白质结构和功能的较少改变。以下是氨基酸的多个分组的实例：1)具有非极性R基团的氨基酸：丙氨酸、缬氨酸、亮氨酸、异亮氨酸、脯氨酸、苯丙氨酸、色氨酸、甲硫氨酸；2)具有不带电的极性R基团的氨基酸：甘氨酸、丝氨酸、苏氨酸、半胱氨酸、酪氨酸、天冬酰胺、谷氨酰胺；3)具有带电的极性R基团的氨基酸(在pH 6.0时带负电)：天冬氨酸、谷氨酸；4)碱性氨基酸(在pH 6.0时带正电)：赖氨酸、精氨酸、组氨酸(在pH6.0)。另一分组可以是具有苯基的那些氨基酸：苯丙氨酸、色氨酸，和酪氨酸。
在某些实施方案中，间隔物源自IgG1、IgG2、IgG3或IgG4，其包含用与未修饰间隔物中存在的氨基酸残基不同的氨基酸残基取代的一个或多个氨基酸残基。所述一个或多个取代的氨基酸残基选自，但不限于位置220、226、228、229、230、233、234、235、234、237、238、239、243、247、267、268、280、290、292、297、298、299、300、305、309、218、326、330、331、332、333、334、336、339处的一个或多个氨基酸残基，或其组合。在该编号体系中，在下文更详细地描述，表1中IgG4(L235E、N297Q)间隔物中的第一个氨基酸是219，并且表1中IgG4(HL-CH3)间隔物中的第一个氨基酸是219，表1中IgG铰链序列和IgG4铰链接头(HL)序列中的第一个氨基酸亦然。
在一些实施方案中，修饰的间隔物源自IgG1、IgG2、IgG3或IgG4，其包含，但不限于一个或多个以下氨基酸残基取代：C220S、C226S、S228P、C229S、P230S、E233P、V234A、L234V、L234F、L234A、L235A、L235E、G236A、G237A、P238S、S239D、F243L、P247I、S267E、H268Q、S280H、K290S、K290E、K290N、R292P、N297A、N297Q、S298A、S298G、S298D、S298V、T299A、Y300L、V305I、V309L、E318A、K326A、K326W、K326E、L328F、A330L、A330S、A331S、P331S、I332E、E333A、E333S、E333S、K334A、A339D、A339Q、P396L，或其组合。
在某些实施方案中，修饰的间隔物源自IgG4区，其包含用与未修饰区域中存在的氨基酸残基不同的氨基酸残基取代的一个或多个氨基酸残基。一个或多个取代的氨基酸残基选自，但不限于位置220、226、228、229、230、233、234、235、234、237、238、239、243、247、267、268、280、290、292、297、298、299、300、305、309、218、326、330、331、332、333、334、336、339上的一个或多个氨基酸残基或其组合。
在一些实施方案中，修饰的间隔物源自IgG4区，其包含，但不限于以下氨基酸残基取代的一个或多个：220S、226S、228P、229S、230S、233P、234A、234V、234F、234A、235A、235E、236A、237A、238S、239D、243L、247I、267E、268Q、280H、290S、290E、290N、292P、297A、297Q、298A、298G、298D、298V、299A、300L、305I、309L、318A、326A、326W、326E、328F、330L、330S、331S、331S、332E、333A、333S、333S、334A、339D、339Q、396L或其组合，其中未修饰间隔物中的氨基酸被所指示位置上的上文鉴定的氨基酸取代。
对于本文讨论的免疫球蛋白中的氨基酸位置，根据EU索引或EU编号体系(Kabatet al.1991Sequences of Proteins of Immunological Interest,5th Ed.,UnitedStates Public Health Service,National Institutes of Health,Bethesda,载体通过引用完整并入)进行编号。EU索引或Kabat或EU编号体系中的EU索引指EU抗体的编号(Edelman et al.1969Proc Natl Acad Sci USA 63:78-85)。
可以使用多种跨膜域。表2包含合适的跨膜域的实例。在存在间隔物域的情况下，跨膜域定位在间隔物域的羧基末端。
表2：跨膜域的实例
Figure GDA0003366579280000101
本文所述的许多CAR包含一个或多个(例如两个)共刺激域。共刺激域位于跨膜域和CD3ζ信号传导域之间。表3包括合适的共刺激域的实例和CD3ζ信号传导域的序列。
表3：CD3ζ域和共刺激域的实例
Figure GDA0003366579280000102
Figure GDA0003366579280000111
附图简述
图1描绘了Her2scFv-IgG4(S228P,L235E,N297Q)-CD28tm-CD28gg-Zeta-T2A-CD19t的氨基酸序列(SEQ ID NO:26)。各个域在序列下按顺序列出，并以交替下划线和非下划线指示。成熟CAR序列(SEQ ID NO:34)不包括GMCSFRa信号肽、T2A跳跃序列或截短的CD19。图3A-8E中以及贯穿整个说明书也称为HER2(EQ)28ζ。图2描绘了Her2scFv-IgG4(S228P,L235E,N297Q)-CD8tm-41BB-Zeta-T2A-CD19t的氨基酸序列(SEQ ID NO:27)。各个域在序列下按顺序列出，并以交替下划线和非下划线指示。成熟CAR序列(SEQ ID NO:35)不包括GMCSFRa信号肽、T2A跳跃序列或截短的CD19。图3A-8E中以及贯穿整个说明书也称为HER2(EQ)BBζ。
图3A-D描绘了HER2特异性CAR构建体和CAR T细胞扩充数据。
图4A-D描绘针对乳腺癌细胞系的HER2-CAR T细胞的体外表征。
图5A-5F描绘了对HER2-CAR T细胞的体外肿瘤活性的研究结果。
图6A-6I描绘了对局部肿瘤内递送的HER2-CAR T细胞的体内抗肿瘤功效的研究结果。
图7A-7D描绘了对人原位(orthotopic)BBM异种移植物模型中HER2-CAR T细胞局部递送的研究结果。
图8A-8D描绘了对HER2-CAR T细胞的室内递送的研究结果。
图9描绘了Her2scFv-CD8hinge
Figure GDA0003366579280000112
CD8tm-41BB-Zeta-T2A-CD19t的氨基酸序列(SEQID NO:28)。各种域在序列下方按顺序列出，并通过交替的下划线和非下划线表示。成熟的CAR序列(SEQ ID NO:36)不包括GMCSFRa信号肽、T2A跳跃序列或截短的CD19。
图10描绘了Her2scFv-IgG4铰链(S228P)-接头-CD8tm-41BB-Zeta-T2A-CD19t的氨基酸序列(SEQ ID NO:29)。各种结构域在序列下方按顺序列出，并由交替的下划线和非下划线表示。成熟的CAR序列(SEQ ID NO:37)不包括GMCSFRa信号肽、T2A跳跃序列或截短的CD19。
图11描绘了Her2scFv-IgG4(S228P)-接头-IgG4CH3-CD8tm-41BB-Zeta-T2A-CD19t的氨基酸序列(SEQ ID NO:30)。各种域在序列下方按顺序列出，并通过交替的下划线和非下划线表示。成熟的CAR序列(SEQ ID NO:38)不包括GMCSFRa信号肽、T2A跳跃序列或截短的CD19。
图12描绘了Her2scFv-IgG4(S228P)-接头-IgG4CH3
Figure GDA0003366579280000121
CD28tm-CD28gg-Zeta-T2A-CD19t(SEQ ID NO:31)的氨基酸序列(SEQ ID NO:31)。各种域在序列下方按顺序列出，并通过交替的下划线和非下划线表示。成熟的CAR序列(SEQ ID NO:39)不包括GMCSFRa信号肽、T2A跳跃序列或截短的CD19。
图13描绘了Her2scFv-接头-CD28tm-CD28gg-Zeta-T2A-CD19t的氨基酸序列(SEQID NO:32)。各种域在序列下方按顺序列出，并通过交替的下划线和非下划线表示。成熟的CAR序列(SEQ ID NO:40)不包括GMCSFRa信号肽、T2A跳跃序列或截短的CD19。
图14描绘了Her2scFv-接头-CD8tm-41BB-Zeta-T2A-CD19t的氨基酸序列(SEQ IDNO:33)。各种域在序列下方按顺序列出，并通过交替的下划线和非下划线表示。成熟的CAR序列(SEQ ID NO:41)不包括GMCSFRa信号肽、T2A跳跃序列或截短的CD19。
图15A-15C描绘了表征具有各种间隔物的某些另外的CAR的研究结果。IgG3(EQ)在图2中；DeltaCh2在图11中；CD8h在图9中；HL在图10中；L在图14中。
图16A-16C显示了检查当将表达各种CAAR的TCM暴露于不表达HER2(MDA-MB-468)、低HER2(231BR)、低HER2(231BRHER2LO)或高HER2(231BRHER2HI)的细胞时产生的CD107a和INFγ的研究结果。
图17A-17D显示了检查用图2(HER2(EQ)BBζ)或图14(HER2(L)BBζ)的CAR的各种细胞系中的PD-1产生和肿瘤细胞杀伤的研究结果。
图18A-18B显示了检查当将表达各种CAAR的TCM暴露于不表达HER2(MDA-MB-468)、低HER2(231BR)、低HER2(231BRHER2LO)或高HER2(231BRHER2HI)的细胞时产生的CD107a和INFγ的研究结果。
发明详述
以下描述了靶向HER2并且可用于治疗HER2表达性乳腺癌以及乳房至脑转移的各种嵌合抗原受体的结构、构建和表征。重要的是，本文所述的CAR可以用于ACT以通过室内或瘤内递送来治疗脑中的HER2表达性肿瘤。
嵌合抗原(CAR)是重组生物分子，其至少含有胞外识别域、跨膜区和胞内信号传导域。因此，术语“抗原”不限于结合抗体的分子，而且指能够特异性结合靶物的任何分子。例如，CAR可以包括特异性结合细胞表面受体的配体。胞外识别域(也称为胞外域或简单地通过其所含的识别元件提及)包含特异性结合存在于靶细胞的细胞表面上的分子的识别元件。跨膜区将CAR锚定在膜上。胞内信号传导域包含来自人CD3复合物的ζ链的信号传导域，并且任选地包含一个或多个共刺激信号传导域。CAR既可以结合抗原，又可以转导T细胞活化，而不依赖于MHC限制。因此，CAR是“普遍的”免疫受体，其可以治疗具有抗原阳性肿瘤的患者群体，而不论其HLA基因型如何。使用表达肿瘤特异性CAR的T淋巴细胞的过继免疫治疗可以是用于治疗癌症的强大治疗策略。
在一些情况下，可以使用其中CAR可读框之后是T2A核糖体跳跃序列和缺少细胞质信号传导尾(在氨基酸323处截短)的截短型CD19(CD19t)的载体来产生本文所述的CAR。在此种排列中，CD19t的共表达提供了惰性非免疫原性表面标志物，其允许对经基因修饰的细胞进行准确测量，并且能够阳性选择经基因修饰的细胞，以及在过继转移后在体内对治疗性T细胞进行有效的细胞追踪和/或成像。CD19t的共表达提供了标志物，其用于使用临床可用的抗体和/或免疫毒素试剂在体内免疫学靶向经转导的细胞以选择性消减治疗性细胞并由此发挥自杀开关的功能。
本文所述的CAR可以通过本领域已知的任何方式产生，尽管优选使用重组DNA技术产生它。方便时，可以通过本领域已知的分子克隆的标准技术(基因组文库筛选、PCR、引物辅助连接、定点诱变等)制备编码嵌合受体几个区域的核酸并将其组装成完整的编码序列。所产生的编码区优选插入表达载体并用于转化合适的表达宿主细胞系，优选T淋巴细胞系，并最优选自体T淋巴细胞系。
可以用CAR表达用的载体转导从患者分离的各种T细胞亚组，包括未选择的PBMC或富集的CD3T细胞或富集的CD3或记忆T细胞亚组。中央记忆T细胞是一种有用的T细胞亚组。可以通过富集CD45RO+/CD62L+细胞，使用例如
Figure GDA0003366579280000141
装置免疫磁性选择表达期望受体的细胞从外周血单个核细胞(PBMC)分离中央记忆T细胞。富含中央记忆T细胞的细胞可以用抗CD3/CD28激活，用例如指导CAR以及截短的人CD19(CD19t)表达的SIN慢病毒载体转导，所述截短的人CD19是用于体内检测和潜在的离体选择两者的非免疫原性表面标志物。活化/遗传修饰的中央记忆T细胞可以用IL-2/IL-15体外扩充，然后冷冻保存。
实施例1：两种HER2-CAR的结构
包含本文描述的HER2 scFv的一种CAR称为Her2scFv-IgG4(L235E，N297Q)-CD28tm-CD28gg-Zeta-T2A-CD19t。此CAR包括多种重要特征，包括：靶向HER2的scFv；以降低Fc受体(FcR)结合的方式在CH2区内的两个位点处突变(L235E；N297Q)的IgG4 Fc区；CD28跨膜域、CD28共刺激域和CD3ζ活化域。图1呈现了此CAR的氨基酸序列，包括用于监测CAR表达的截短的CD19序列的序列和允许在不融合截短的CD19序列的情况下产生CAR的T2A核糖体跳跃序列。如图2所示，未成熟的CAR包括：GMCSFR信号肽、HER2scFv、起间隔物作用的IgG4、CD8跨膜区、包含LL至GG序列改变的4-1BB共刺激域、三Gly序列、CD3 Zeta刺激域。转录物也编码T2A核糖体序列和截短的CD19序列，其不是CAR蛋白序列的一部分。成熟CAR与未成熟CAR相同，但缺少GMCSF信号肽。
实施例2：用于表达HER2-特异性CAR T细胞的epHIV7的构建和结构
epHIV7载体是可以用于表达HER2特异性CAR的载体。它从pHIV7载体产生。重要的是，此种载体使用人EFI启动子来驱动CAR的表达。载体的5’和3’序列都源自pv653RSN，如先前源自HXBc2原病毒。聚嘌呤段(tract)DNA瓣(flap)序列(cPPT)源自NIH AIDS试剂保藏所(Reagent Repository)的HIV-I株pNL4-3。先前描述了土拨鼠转录后调控元件(WPRE)序列。
如下进行pHIV7的构建。简言之，如下将含有来自gag-pol的653bp加上5’和3’长末端重复(LTR)以及居间的SL3-新霉素磷酸转移酶基因(Neo)的pv653RSN亚克隆到pBluescript中：在步骤1中，从5’LTR到rev-应答元件(RRE)的序列制备p5’HIV-I 5I，然后通过除去TATA盒上游的序列修饰5’LTR，并首先连接到CMV增强子，然后连接到SV40复制起点(p5’HIV-2)。在步骤2中，在将3’LTR克隆到pBluescript中以制备p3’HIV-1后，在3’LTR增强子/启动子中进行400-bp缺失以除去HIV U3中的顺式调控元件并形成p3’HIV-2。在步骤3中，连接从p5’HIV-3和p3’HIV-2分离的片段以制备pHIV-3。在步骤4中，通过除去额外的上游HIV序列来进一步修饰p3’HIV-2以产生p3’HIV-3，并且将含有WPRE的600-bp BamHI-SalI片段添加到p3’HIV-3中以制备p3’HIV-4。在步骤5中，通过PCR将pHIV-3RRE的大小减小，并连接到来自pHIV-3的5’片段(未显示)以及连接到p3’HIV-4，以制备pHIV-6。在步骤6中，从pNL4-3扩增含有来自HIV-1pNL4-3(55)的cPPT DNA瓣序列的190-bp BglII-BamHI片段，并将其置于pHIV6中的RRE和WPRE序列之间以制备pHIV-7。使用该亲本质粒pHIV7-GFP(GFP，绿色荧光蛋白)，使用4-质粒系统包装亲本载体。
包装信号psiψ是将病毒基因组有效包装到载体中所需的。RRE和WPRE增强RNA转录物转运和转基因表达。与WPRE组合的瓣序列已被证明增强哺乳动物细胞中慢病毒载体的转导效率。
产生病毒载体所需的辅助功能被分到三个分开的质粒中以降低通过重组产生有复制能力的慢病毒的可能性：1)pCgp编码病毒载体装配所需的gag/pol蛋白；2)pCMV-Rev2编码Rev蛋白，其作用于RRE序列以帮助运输病毒基因组高效包装；和3)pCMV-G编码病毒载体的感染性所需的水泡-口炎病毒(VSV)的糖蛋白。
在pHIV7编码的载体基因组和辅助质粒之间具有最小的DNA序列同源性。同源性区域包括位于pCgp辅助质粒的gag/pol序列中的约600个核苷酸的包装信号区域；所有三种辅助质粒中的CMV启动子序列；和辅助质粒pCgp中的RRE序列。由于这些区域中的同源性而可以产生具有复制能力的重组病毒是非常不可能的，因为其需要多次重组事件。此外，任何产生的重组体都将缺少慢病毒复制所需的功能性LTR和tat序列。
CMV启动子被EF1α-HTLV启动子(EF1p)替换，并且新质粒被命名为epHIV7。EF1p具有563bp并在切除CMV启动子后使用NruI和NheI引入epHIV7。
已经从该系统中除去了慢病毒基因组，排除野生型病毒的致病性所必需的且靶细胞的生产性感染所需的gag/pol和rev。此外，载体构建体不含完整的3’LTR启动子，因此在靶定细胞中得到的表达的且反转录的DNA原病毒基因组将具有无活性的LTR。由于此种设计，无HIV-I衍生的序列将从原病毒转录，并且仅治疗序列将从它们各自的启动子表达。SIN载体中LTR启动子活性的除去显著降低宿主基因的无意激活的可能性。
实施例3：用于转导患者T细胞的载体的制备
可以如下制备患者T细胞的转导用的载体。对于表达CAR的每种质粒，表达CAR和任选地标志物，诸如截短型CD19；2)pCgp；3)pCMV-G；和4)pCMV-Rev2)，产生种子库，其用于接种发酵罐以产生足够量的质粒DNA。在用于产生慢病毒载体之前，测试质粒DNA的身份、无菌性和内毒素。
简言之，从293T工作细胞(WCB)中扩充细胞，所述293T工作细胞经测试证实无菌并且没有病毒污染。将来自293T WCB的一小瓶293T细胞解冻。培养并扩充细胞直到存在足够数量的细胞来铺板适当数量的10层细胞厂(CF)，用于载体生产和细胞培养维持。细胞的单培养(single train)可用于生产。
在最多10CF的亚批次中产生慢病毒载体。可以在同一周内产生两个亚批次，导致产生约20L慢病毒上清液/周。在下游加工阶段期间合并从全部亚批次产生的材料，以产生一批产物。将293T细胞铺在293T培养基(含有10％FBS的DMEM)中的CF中。将厂(factory)放置在37℃培养箱中并水平地放平以获得CF的所有层上细胞的均匀分布。两天后，使用CaPO4方法用上述四种慢病毒质粒转染细胞，所述方法涉及Tris:EDTA、2M CaCh、2X HBS和四种DNA质粒的混合物。转染后第3天，收集含有分泌的慢病毒载体的上清液，纯化并浓缩。从CF中除去上清液后，从每个CF收集最终生产的细胞。将来自每个厂的细胞用胰蛋白酶消化处理并通过离心收集。将细胞重悬浮于冷冻培养基中并冷冻保存。这些细胞后来用于有复制能力的慢病毒(RCL)测试。
为了纯化和配制载体，通过膜过滤澄清粗制上清液以除去细胞碎片。通过内切核酸酶消化
Figure GDA0003366579280000161
降解宿主细胞DNA和残留质粒DNA。使用0.45μm过滤器对病毒上清液澄清细胞碎片。将澄清的上清液收集到预先称重的已经加入有
Figure GDA0003366579280000162
(终浓度50U/mL)的容器中。残留质粒DNA和宿主基因组DNA的核酸内切酶消化在37℃下进行6小时。使用核酸内切酶处理的上清液的初始切向流超滤(TFF)浓缩从粗上清液中除去残留的低分子量组分，而将病毒浓缩约20倍。将澄清的核酸内切酶处理的病毒上清液在使流量速率最大化的情况下以设计为维持约4,000秒-1或更小的剪切速率的流速循环通过NMWCO为500kD的中空纤维筒。在浓缩过程期间启动经核酸酶处理的上清液的渗滤以维持筒性能。使用PBS中4％乳糖作为渗滤缓冲液，建立了80％的渗透物置换率。使病毒上清液达到目标体积，代表粗制上清液的20倍浓缩，并渗滤再继续4个交换体积，渗透物置换率为100％。
通过使用高速离心技术实现病毒产物的进一步浓缩。使用Sorvall RC-26plus离心机以6000RPM(6,088RCF)在6℃将每个亚批次的慢病毒沉淀16-20小时。然后将来自每个亚批次的病毒团粒在PBS中具有4％乳糖的50mL体积中重建。该缓冲液中的重建的团粒代表病毒制备物的最终配制剂。整个载体浓缩过程导致约200倍的体积减少。在所有亚批次完成之后，然后将材料置于-80℃，同时对来自每个亚批次的样品测试无菌性。确认样品无菌性后，将亚批次在37℃在频繁搅拌下迅速融化。然后将材料合并，并且手动在II类A/B3生物安全柜中等分取样。使用在无菌USP 6类、外螺纹的O形环冷冻管中的1mL浓缩慢病毒的填充构造。
为了确保慢病毒载体制备物的纯度，对其测试残留宿主DNA污染物和残留宿主和质粒DNA的转移。在其他测试中，通过RT-PCR评估载体身份以确保存在正确的载体。
实施例4：适合在ACT中使用的T细胞的制备
若要使用Tcm表达CAR，则可以如下制备合适的患者细胞。首先，通过白细胞分离术(leukopheresis)从患者获得T淋巴细胞，并且将合适的同种异体或自体T细胞亚组(例如中央记忆T细胞(TCM))遗传改变以表达CAR，然后通过任何临床上可接受的手段将其施用回患者以实现抗癌治疗。
可以如下产生合适的TCM。从同意的研究参与者获得的单采血液成分术产物用ficoll处理，清洗并温育过夜。然后使用GMP级抗-CD14，抗-CD25和抗-CD45RA试剂(Miltenyi Biotec)和CliniMACSTM分离装置消减细胞中的单核细胞、调节性T细胞和幼稚
Figure GDA0003366579280000171
T细胞群体。消减后，在CliniMACSTM分离装置上使用DREG56-生物素(COH临床级)和抗生物素微珠(MiltenyiBiotec)针对CD62L+TCM细胞富集的阴性级分细胞。
富集后，在完全X-Vivo15加50IU/mL IL-2和0.5ng/mL IL-15中配制TCM细胞，并将其转移至Teflon细胞培养袋中，其中用Dynal ClinExTM Vivo CD3/CD28珠刺激它们。刺激后长达五天，以感染复数(MOI)为1.0至0.3用表达期望的CAR的慢病毒载体转导细胞。在根据细胞扩增所需要加入完全X-Vivo15和IL-2和IL-15细胞因子的情况下将培养物维持长达42天(将细胞密度保持在3x105和2x106活细胞/mL之间，并且培养的每周一、周三和周五补充细胞因子)。细胞在21天内在这些条件下通常扩增至约109个细胞。在培养期结束时收集细胞，清洗两次并配制在临床级冷冻保存培养基(Cryostore CS5,BioLife Solutions)中。
在T细胞输注的当天，将冷冻保存并释放的产物融化，清洗并配制用于再输注。将含有释放的细胞产物的冷冻保存的小瓶从液氮储存中取出，融化，冷却并用PBS/2％人血清白蛋白(HSA)清洗缓冲液清洗。离心后，除去上清液并将细胞重悬浮于不含防腐剂的生理盐水(PFNS)/2％HSA输注稀释剂中。取出样品用于质量控制测试。
实施例5：靶向HER2的CAR的表达
图3A是图1和图2中描绘的两种HER2-特异性CAR构建体的示意图。在HER2(EQ)28ξ中，通过经修饰的IgG4 Fc接头(双重突变体，L235E；N297Q)将scFv栓系到膜，含有CD28跨膜域、胞内CD28共刺激域和细胞溶解性CD3ζ域。T2A跳跃序列将CAR与用于细胞追踪的截短型CD19(CD19t)蛋白分开。HER2(EQ)BBξ是相似的，只是共刺激域是4-1BB而不是CD28并且跨膜域是CD8跨膜域而不是CD28跨膜域。用表达HER2(EQ)28ξ或HER2(EQ)BBξ的慢病毒载体转染人中央记忆(TCM)细胞。图3B描绘了人TCM表面表型的代表性FACS数据。图3C描绘了用表达HER2(EQ)28ξ或HER2(EQ)BBξ的慢病毒载体转染的TCM中CD19和蛋白L表达的测定结果。从这些结果可以看出，对于这两种CAR，如通过CD19表达评估的转染效率是相似的。然而，蛋白L表达对于HER2(EQ)BBξ比对于HER2(EQ)28ξ更低，提示HER2(EQ)BBξCAR的稳定性小于HER2(EQ)BBξ。细胞扩充的分析(图3D)显示这两种CAR都不干扰T细胞扩充。
实施例6：针对各种乳腺癌细胞系的HER2-CAR T细胞的体外表征
使用多种乳腺癌细胞系，包括HER2阴性系(LCL淋巴瘤，MDA-MB-468，U87神经胶质瘤)、低HER2表达系(MDA-MB-361，231BR)和高HER2表达系(SKBR3、BT474、BBM1)来表征HER2(EQ)28和HER2(EQ)BBξ。图4A描绘了这些系中每种的HER2表达水平。使用流式细胞术(对CAR+T细胞门控)表征与MDA-MB-361肿瘤细胞(低HER2表达)或BBM1肿瘤细胞(高HER2表达)共培养5小时后模拟物(未转导的)、HER2(EQ)28ξ或HER2(EQ)BBξCAR T细胞中的CD107a脱粒和IFNγ产生。此种分析的结果在图4B中呈现。对其他乳腺癌细胞系进行了类似的研究，并且结果总结在图4C中。通过ELISA测量在用重组HER2蛋白或肿瘤靶物培养24小时后通过HER2-CAR T细胞生产的IFNγ产量，并且该分析的结果显示在图4D中。
实施例7：体外抗肿瘤活性
使用流式细胞术来评估模拟物(未转导的)、HER2(EQ)28ξ或HER2(EQ)BBξCAR T细胞与肿瘤靶物共培养72小时后的肿瘤细胞杀伤。该分析的结果在图5A中呈现。测量了与HER2阴性MDA-MB-468或HER2阳性BBM1细胞共培养72小时后总CAR T细胞中的PD-1和LAG-3诱导，并且该分析的结果呈现于图5B中。在与HER2阴性(LCL淋巴瘤，MDA-MB-468，U87神经胶质瘤)，低HER2表达(MDA-MB-361，231BR)或高HER2表达(SKBR3，BT474，BBM1)的肿瘤靶物共培养72小时后，CD8+CAR T细胞中的PD-1诱导，并且此分析的结果在图5C中呈现。这些研究提示了HER2(EQ)BBξ比HER2(EQ)28ξ引起更低的PD-1诱导。对HER2(EQ)28ξ或HER2(EQ)BBξCAR T细胞两者测量到具有范围为0.25:1至2:1的效应器:肿瘤(E:T)比率的肿瘤细胞杀伤。该分析的结果在图5D中呈现，其显示了HER2(EQ)28ξ或HER2(EQ)BBξ两者在体外肿瘤细胞杀伤中是有效的。通过流式细胞术测量在与MDA-MB-468或BBMI细胞共培养72小时后HER2-CART细胞的CFSE增殖。此分析的结果在图5E中呈现，其显示了HER2(EQ)BBξCAR T细胞比HER2(EQ)28ξCAR T细胞增殖得更多。
实施例8：体内抗肿瘤活性
在患者衍生的乳房至脑转移模型中评估瘤内递送的HER2 CAR T细胞的活性。图6A-6C是肿瘤的H&E染色。通过用模拟物(未转导的)或HER2(EQ)BBξCAR T细胞直接注射到肿瘤中来治疗小鼠。图6D-6F描述了肿瘤的光学成像结果，而图6G-6I是在肿瘤注射后第3天、第8天或第14天时局部治疗的小鼠的Kaplan-Meier存活曲线。这些研究显示当直接注射到肿瘤中时，HER2(EQ)BBξCAR T细胞具有有力的体内抗肿瘤效力。
为了评估人乳腺至脑转移异种移植物模型中的抗肿瘤功效，将BBM1细胞(0.2M)或BT474(0.15M)颅内注射到NSG小鼠中。在肿瘤注射后的第8天，肿瘤内注射HER2(EQ)28ξ、HER2(EQ)BBξ或模拟物(未转导的)T细胞(0.5M)。通过基于萤光素酶的光学成像监测BBMI(图7A)和BT474(图7B)。图7C和图7D中呈现了Kaplan Meier曲线。
乳腺至脑转移的人患者来源的原位异种移植物模型也用于评估HER2(EQ)28ξ或HER2(EQ)BBξCAR T细胞。图8A显示了通过立体定向注射BBM1细胞(0.2M)和室内T细胞递送的肿瘤植入区。肿瘤染色描绘于图8B中。在肿瘤注射后第14天，瘤内注射HER2(EQ)28ξ、HER2(EQ)BBξ或模拟物(未转导的)T细胞(0.5M)。图8C呈现了每个治疗组的通量平均值，而图8D呈现了每个治疗组的Kaplan Meier存活曲线。
实施例9：靶向HER2的另外CAR
图9-14描绘了具有不同接头的各种CAR的氨基酸序列。具体地，CAR在靶向HER2的scFv和跨膜域之间的部分的序列和长度上不同。跨膜域是CD8，CD28或CD28gg。共刺激域是4-1BB或CD28。全部都有CD3ζ刺激域。在每种情况下，T2A跳跃序列将CAR与用于细胞追踪的截短型CD19(CD19t)蛋白分开。
图15A示意性描绘了各种HER2 CAR，除了HER2 scFv和CD8跨膜域之间的部分的序列和长度以外，它们是相同的。全部包括4-1BB共刺激域，随后是CD3ζ刺激域。图15B描绘了用表达所示CAR的慢病毒载体转染的TCM中的CD19和蛋白L表达的测定结果。从这些结果可以看出，对于两种CAR，如通过CD19表达评估的转染效率是相似的。然而，蛋白L表达对于HER2(EQ)BBξ比对于HER2(EQ)28ξ低，提示HER2(EQ)BBξCAR的稳定性小于HER2(EQ)BBξ。对细胞扩充的分析(图15C)显示CAR无一干扰T细胞扩充，图16-18显示了另外的研究结果，其显示具有非常短的间隔物的CAR(图14)对于表达高水平HER2的CAR具有相对选择性。此类CAR可以可用于治疗表达HER2的癌症，其中期望剩下表达比癌细胞低的HER2水平的细胞。
序列表
<110> Priceman, Saul J.
Forman, Stephen J.
Brown, Christine E.
<120> 靶向HER2的嵌合抗原受体
<130> 40056-0021WO1
<140> US 15/773,754
<141> 2018-05-04
<150> PCT/US2016/060724
<151> 2016-11-04
<150> US 62/251,052
<151> 2015-11-04
<160> 42
<170> PatentIn version 3.5
<210> 1
<211> 245
<212> PRT
<213> 人
<220>
<221> 变体
<222> (1)..(245)
<223> 靶向HER2的scEv
<400> 1
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Glu Val Gln
115 120 125
Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
130 135 140
Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His
145 150 155 160
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile
165 170 175
Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg
180 185 190
Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met
195 200 205
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp
210 215 220
Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu
225 230 235 240
Val Thr Val Ser Ser
245
<210> 2
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> 接头
<400> 2
Gly Gly Gly Ser Ser Gly Gly Gly Ser Gly
1 5 10
<210> 3
<211> 12
<212> PRT
<213> 人工序列
<220>
<223> 突变的IgG4铰链区
<400> 3
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro
1 5 10
<210> 4
<211> 12
<212> PRT
<213> 人
<220>
<221> 变体
<222> (1)..(12)
<400> 4
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro
1 5 10
<210> 5
<211> 22
<212> PRT
<213> 人工序列
<220>
<223> 嵌合IgG4 突变的铰链和接头
<400> 5
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Gly Gly Gly Ser
1 5 10 15
Ser Gly Gly Gly Ser Gly
20
<210> 6
<211> 39
<212> PRT
<213> 人
<400> 6
Ile Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn
1 5 10 15
Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu
20 25 30
Phe Pro Gly Pro Ser Lys Pro
35
<210> 7
<211> 48
<212> PRT
<213> 人
<400> 7
Ala Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro
1 5 10 15
Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro
20 25 30
Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp
35 40 45
<210> 8
<211> 45
<212> PRT
<213> 人
<400> 8
Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala
1 5 10 15
Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly
20 25 30
Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp
35 40 45
<210> 9
<211> 129
<212> PRT
<213> 人工序列
<220>
<223> 嵌合IgG4 铰链 +
<400> 9
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Gly Gly Gly Ser
1 5 10 15
Ser Gly Gly Gly Ser Gly Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
20 25 30
Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
35 40 45
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
50 55 60
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
65 70 75 80
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys
85 90 95
Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu
100 105 110
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
115 120 125
Lys
<210> 10
<211> 228
<212> PRT
<213> 人工序列
<220>
<223> 突变的IgG4
<400> 10
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe
1 5 10 15
Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
20 25 30
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
35 40 45
Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val
50 55 60
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Ser Thr
65 70 75 80
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
85 90 95
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser
100 105 110
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
115 120 125
Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val
130 135 140
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
145 150 155 160
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
165 170 175
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr
180 185 190
Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val
195 200 205
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
210 215 220
Ser Leu Gly Lys
225
<210> 11
<211> 229
<212> PRT
<213> 人工序列
<220>
<223> 突变的IgG4
<400> 11
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe
1 5 10 15
Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
20 25 30
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
35 40 45
Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val
50 55 60
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Gln Ser
65 70 75 80
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
85 90 95
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser
100 105 110
Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
115 120 125
Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln
130 135 140
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
145 150 155 160
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
165 170 175
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu
180 185 190
Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser
195 200 205
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
210 215 220
Leu Ser Leu Gly Lys
225
<210> 12
<211> 107
<212> PRT
<213> 人
<400> 12
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu
1 5 10 15
Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
20 25 30
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
35 40 45
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
50 55 60
Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly
65 70 75 80
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
85 90 95
Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
100 105
<210> 13
<211> 21
<212> PRT
<213> 人
<400> 13
Leu Cys Tyr Leu Leu Asp Gly Ile Leu Phe Ile Tyr Gly Val Ile Leu
1 5 10 15
Thr Ala Leu Phe Leu
20
<210> 14
<211> 27
<212> PRT
<213> 人
<400> 14
Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu
1 5 10 15
Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val
20 25
<210> 15
<211> 28
<212> PRT
<213> 人
<400> 15
Met Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser
1 5 10 15
Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val
20 25
<210> 16
<211> 22
<212> PRT
<213> 人
<400> 16
Met Ala Leu Ile Val Leu Gly Gly Val Ala Gly Leu Leu Leu Phe Ile
1 5 10 15
Gly Leu Gly Ile Phe Phe
20
<210> 17
<211> 21
<212> PRT
<213> 人
<400> 17
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu
1 5 10 15
Ser Leu Val Ile Thr
20
<210> 18
<211> 23
<212> PRT
<213> 人
<400> 18
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu
1 5 10 15
Ser Leu Val Ile Thr Leu Tyr
20
<210> 19
<211> 24
<212> PRT
<213> 人
<400> 19
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu
1 5 10 15
Ser Leu Val Ile Thr Leu Tyr Cys
20
<210> 20
<211> 28
<212> PRT
<213> 人
<400> 20
Ile Ile Ser Phe Phe Leu Ala Leu Thr Ser Thr Ala Leu Leu Leu Phe
1 5 10 15
Leu Leu Phe Phe Leu Thr Leu Arg Phe Ser Val Val
20 25
<210> 21
<211> 112
<212> PRT
<213> 人
<400> 21
Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly
1 5 10 15
Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr
20 25 30
Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys
35 40 45
Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys
50 55 60
Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg
65 70 75 80
Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala
85 90 95
Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
100 105 110
<210> 22
<211> 41
<212> PRT
<213> 人
<400> 22
Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr
1 5 10 15
Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro
20 25 30
Pro Arg Asp Phe Ala Ala Tyr Arg Ser
35 40
<210> 23
<211> 41
<212> PRT
<213> 人工序列
<220>
<223> 突变的人的
<400> 23
Arg Ser Lys Arg Ser Arg Gly Gly His Ser Asp Tyr Met Asn Met Thr
1 5 10 15
Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro
20 25 30
Pro Arg Asp Phe Ala Ala Tyr Arg Ser
35 40
<210> 24
<211> 42
<212> PRT
<213> 人
<400> 24
Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met
1 5 10 15
Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe
20 25 30
Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu
35 40
<210> 25
<211> 42
<212> PRT
<213> 人
<400> 25
Ala Leu Tyr Leu Leu Arg Arg Asp Gln Arg Leu Pro Pro Asp Ala His
1 5 10 15
Lys Pro Pro Gly Gly Gly Ser Phe Arg Thr Pro Ile Gln Glu Glu Gln
20 25 30
Ala Asp Ala His Ser Thr Leu Ala Lys Ile
35 40
<210> 26
<211> 1027
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-IgG4(L235E,
N297Q)-CD28tm-CD28gg-Zeta-T2A-CD19t
<400> 26
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
1 5 10 15
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Asp Val Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
His Tyr Thr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
115 120 125
Lys Gly Ser Thr Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
145 150 155 160
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile
165 170 175
Lys Asp Thr Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
180 185 190
Glu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala
195 200 205
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn
210 215 220
Thr Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
225 230 235 240
Tyr Tyr Cys Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Ser Lys Tyr Gly
260 265 270
Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser
275 280 285
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
290 295 300
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro
305 310 315 320
Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
325 330 335
Lys Thr Lys Pro Arg Glu Glu Gln Phe Gln Ser Thr Tyr Arg Val Val
340 345 350
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
355 360 365
Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr
370 375 380
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
385 390 395 400
Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
405 410 415
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
420 425 430
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
435 440 445
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser
450 455 460
Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
465 470 475 480
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
485 490 495
Met Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser
500 505 510
Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg
515 520 525
Ser Arg Gly Gly His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro
530 535 540
Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe
545 550 555 560
Ala Ala Tyr Arg Ser Gly Gly Gly Arg Val Lys Phe Ser Arg Ser Ala
565 570 575
Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu
580 585 590
Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly
595 600 605
Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu
610 615 620
Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser
625 630 635 640
Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly
645 650 655
Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu
660 665 670
His Met Gln Ala Leu Pro Pro Arg Leu Glu Gly Gly Gly Glu Gly Arg
675 680 685
Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Arg
690 695 700
Met Pro Pro Pro Arg Leu Leu Phe Phe Leu Leu Phe Leu Thr Pro Met
705 710 715 720
Glu Val Arg Pro Glu Glu Pro Leu Val Val Lys Val Glu Glu Gly Asp
725 730 735
Asn Ala Val Leu Gln Cys Leu Lys Gly Thr Ser Asp Gly Pro Thr Gln
740 745 750
Gln Leu Thr Trp Ser Arg Glu Ser Pro Leu Lys Pro Phe Leu Lys Leu
755 760 765
Ser Leu Gly Leu Pro Gly Leu Gly Ile His Met Arg Pro Leu Ala Ile
770 775 780
Trp Leu Phe Ile Phe Asn Val Ser Gln Gln Met Gly Gly Phe Tyr Leu
785 790 795 800
Cys Gln Pro Gly Pro Pro Ser Glu Lys Ala Trp Gln Pro Gly Trp Thr
805 810 815
Val Asn Val Glu Gly Ser Gly Glu Leu Phe Arg Trp Asn Val Ser Asp
820 825 830
Leu Gly Gly Leu Gly Cys Gly Leu Lys Asn Arg Ser Ser Glu Gly Pro
835 840 845
Ser Ser Pro Ser Gly Lys Leu Met Ser Pro Lys Leu Tyr Val Trp Ala
850 855 860
Lys Asp Arg Pro Glu Ile Trp Glu Gly Glu Pro Pro Cys Val Pro Pro
865 870 875 880
Arg Asp Ser Leu Asn Gln Ser Leu Ser Gln Asp Leu Thr Met Ala Pro
885 890 895
Gly Ser Thr Leu Trp Leu Ser Cys Gly Val Pro Pro Asp Ser Val Ser
900 905 910
Arg Gly Pro Leu Ser Trp Thr His Val His Pro Lys Gly Pro Lys Ser
915 920 925
Leu Leu Ser Leu Glu Leu Lys Asp Asp Arg Pro Ala Arg Asp Met Trp
930 935 940
Val Met Glu Thr Gly Leu Leu Leu Pro Arg Ala Thr Ala Gln Asp Ala
945 950 955 960
Gly Lys Tyr Tyr Cys His Arg Gly Asn Leu Thr Met Ser Phe His Leu
965 970 975
Glu Ile Thr Ala Arg Pro Val Leu Trp His Trp Leu Leu Arg Thr Gly
980 985 990
Gly Trp Lys Val Ser Ala Val Thr Leu Ala Tyr Leu Ile Phe Cys Leu
995 1000 1005
Cys Ser Leu Val Gly Ile Leu His Leu Gln Arg Ala Leu Val Leu
1010 1015 1020
Arg Arg Lys Arg
1025
<210> 27
<211> 1021
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合
Her2scFv-IgG4(L235E,N297Q)-CD8tm-41BB-Zeta-T2A-CD19t
<400> 27
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
1 5 10 15
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Asp Val Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
His Tyr Thr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
115 120 125
Lys Gly Ser Thr Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
145 150 155 160
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile
165 170 175
Lys Asp Thr Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
180 185 190
Glu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala
195 200 205
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn
210 215 220
Thr Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
225 230 235 240
Tyr Tyr Cys Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Ser Lys Tyr Gly
260 265 270
Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser
275 280 285
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
290 295 300
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro
305 310 315 320
Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
325 330 335
Lys Thr Lys Pro Arg Glu Glu Gln Phe Gln Ser Thr Tyr Arg Val Val
340 345 350
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
355 360 365
Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr
370 375 380
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
385 390 395 400
Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
405 410 415
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
420 425 430
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
435 440 445
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser
450 455 460
Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
465 470 475 480
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
485 490 495
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu
500 505 510
Ser Leu Val Ile Thr Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe
515 520 525
Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly
530 535 540
Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Gly
545 550 555 560
Gly Gly Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln
565 570 575
Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu
580 585 590
Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly
595 600 605
Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu
610 615 620
Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly
625 630 635 640
Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser
645 650 655
Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro
660 665 670
Pro Arg Leu Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys
675 680 685
Gly Asp Val Glu Glu Asn Pro Gly Pro Arg Met Pro Pro Pro Arg Leu
690 695 700
Leu Phe Phe Leu Leu Phe Leu Thr Pro Met Glu Val Arg Pro Glu Glu
705 710 715 720
Pro Leu Val Val Lys Val Glu Glu Gly Asp Asn Ala Val Leu Gln Cys
725 730 735
Leu Lys Gly Thr Ser Asp Gly Pro Thr Gln Gln Leu Thr Trp Ser Arg
740 745 750
Glu Ser Pro Leu Lys Pro Phe Leu Lys Leu Ser Leu Gly Leu Pro Gly
755 760 765
Leu Gly Ile His Met Arg Pro Leu Ala Ile Trp Leu Phe Ile Phe Asn
770 775 780
Val Ser Gln Gln Met Gly Gly Phe Tyr Leu Cys Gln Pro Gly Pro Pro
785 790 795 800
Ser Glu Lys Ala Trp Gln Pro Gly Trp Thr Val Asn Val Glu Gly Ser
805 810 815
Gly Glu Leu Phe Arg Trp Asn Val Ser Asp Leu Gly Gly Leu Gly Cys
820 825 830
Gly Leu Lys Asn Arg Ser Ser Glu Gly Pro Ser Ser Pro Ser Gly Lys
835 840 845
Leu Met Ser Pro Lys Leu Tyr Val Trp Ala Lys Asp Arg Pro Glu Ile
850 855 860
Trp Glu Gly Glu Pro Pro Cys Val Pro Pro Arg Asp Ser Leu Asn Gln
865 870 875 880
Ser Leu Ser Gln Asp Leu Thr Met Ala Pro Gly Ser Thr Leu Trp Leu
885 890 895
Ser Cys Gly Val Pro Pro Asp Ser Val Ser Arg Gly Pro Leu Ser Trp
900 905 910
Thr His Val His Pro Lys Gly Pro Lys Ser Leu Leu Ser Leu Glu Leu
915 920 925
Lys Asp Asp Arg Pro Ala Arg Asp Met Trp Val Met Glu Thr Gly Leu
930 935 940
Leu Leu Pro Arg Ala Thr Ala Gln Asp Ala Gly Lys Tyr Tyr Cys His
945 950 955 960
Arg Gly Asn Leu Thr Met Ser Phe His Leu Glu Ile Thr Ala Arg Pro
965 970 975
Val Leu Trp His Trp Leu Leu Arg Thr Gly Gly Trp Lys Val Ser Ala
980 985 990
Val Thr Leu Ala Tyr Leu Ile Phe Cys Leu Cys Ser Leu Val Gly Ile
995 1000 1005
Leu His Leu Gln Arg Ala Leu Val Leu Arg Arg Lys Arg
1010 1015 1020
<210> 28
<211> 843
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-CD8铰链-CD8tm-41BB-Zeta-T2A-CD19t
<400> 28
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
1 5 10 15
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Asp Val Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
His Tyr Thr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
115 120 125
Lys Gly Ser Thr Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
145 150 155 160
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile
165 170 175
Lys Asp Thr Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
180 185 190
Glu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala
195 200 205
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn
210 215 220
Thr Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
225 230 235 240
Tyr Tyr Cys Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Lys Pro Thr Thr
260 265 270
Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln
275 280 285
Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala
290 295 300
Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala
305 310 315 320
Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr
325 330 335
Gly Gly Gly Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln
340 345 350
Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser
355 360 365
Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Gly Gly Gly
370 375 380
Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly
385 390 395 400
Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr
405 410 415
Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys
420 425 430
Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys
435 440 445
Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg
450 455 460
Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala
465 470 475 480
Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
485 490 495
Leu Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp
500 505 510
Val Glu Glu Asn Pro Gly Pro Arg Met Pro Pro Pro Arg Leu Leu Phe
515 520 525
Phe Leu Leu Phe Leu Thr Pro Met Glu Val Arg Pro Glu Glu Pro Leu
530 535 540
Val Val Lys Val Glu Glu Gly Asp Asn Ala Val Leu Gln Cys Leu Lys
545 550 555 560
Gly Thr Ser Asp Gly Pro Thr Gln Gln Leu Thr Trp Ser Arg Glu Ser
565 570 575
Pro Leu Lys Pro Phe Leu Lys Leu Ser Leu Gly Leu Pro Gly Leu Gly
580 585 590
Ile His Met Arg Pro Leu Ala Ile Trp Leu Phe Ile Phe Asn Val Ser
595 600 605
Gln Gln Met Gly Gly Phe Tyr Leu Cys Gln Pro Gly Pro Pro Ser Glu
610 615 620
Lys Ala Trp Gln Pro Gly Trp Thr Val Asn Val Glu Gly Ser Gly Glu
625 630 635 640
Leu Phe Arg Trp Asn Val Ser Asp Leu Gly Gly Leu Gly Cys Gly Leu
645 650 655
Lys Asn Arg Ser Ser Glu Gly Pro Ser Ser Pro Ser Gly Lys Leu Met
660 665 670
Ser Pro Lys Leu Tyr Val Trp Ala Lys Asp Arg Pro Glu Ile Trp Glu
675 680 685
Gly Glu Pro Pro Cys Val Pro Pro Arg Asp Ser Leu Asn Gln Ser Leu
690 695 700
Ser Gln Asp Leu Thr Met Ala Pro Gly Ser Thr Leu Trp Leu Ser Cys
705 710 715 720
Gly Val Pro Pro Asp Ser Val Ser Arg Gly Pro Leu Ser Trp Thr His
725 730 735
Val His Pro Lys Gly Pro Lys Ser Leu Leu Ser Leu Glu Leu Lys Asp
740 745 750
Asp Arg Pro Ala Arg Asp Met Trp Val Met Glu Thr Gly Leu Leu Leu
755 760 765
Pro Arg Ala Thr Ala Gln Asp Ala Gly Lys Tyr Tyr Cys His Arg Gly
770 775 780
Asn Leu Thr Met Ser Phe His Leu Glu Ile Thr Ala Arg Pro Val Leu
785 790 795 800
Trp His Trp Leu Leu Arg Thr Gly Gly Trp Lys Val Ser Ala Val Thr
805 810 815
Leu Ala Tyr Leu Ile Phe Cys Leu Cys Ser Leu Val Gly Ile Leu His
820 825 830
Leu Gln Arg Ala Leu Val Leu Arg Arg Lys Arg
835 840
<210> 29
<211> 814
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-铰链-接头-CD8tm-41BB-Zeta-T2A-CD19t
<400> 29
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
1 5 10 15
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Asp Val Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
His Tyr Thr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
115 120 125
Lys Gly Ser Thr Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
145 150 155 160
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile
165 170 175
Lys Asp Thr Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
180 185 190
Glu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala
195 200 205
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn
210 215 220
Thr Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
225 230 235 240
Tyr Tyr Cys Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Ser Lys Tyr Gly
260 265 270
Pro Pro Cys Pro Pro Cys Pro Gly Gly Gly Ser Ser Gly Gly Gly Ser
275 280 285
Gly Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu
290 295 300
Leu Ser Leu Val Ile Thr Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile
305 310 315 320
Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp
325 330 335
Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu
340 345 350
Gly Gly Gly Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr
355 360 365
Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg
370 375 380
Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met
385 390 395 400
Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu
405 410 415
Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys
420 425 430
Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu
435 440 445
Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu
450 455 460
Pro Pro Arg Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys
465 470 475 480
Gly Asp Val Glu Glu Asn Pro Gly Pro Arg Leu Met Pro Pro Pro Arg
485 490 495
Leu Leu Phe Phe Leu Leu Phe Leu Thr Pro Met Glu Val Arg Pro Glu
500 505 510
Glu Pro Leu Val Val Lys Val Glu Glu Gly Asp Asn Ala Val Leu Gln
515 520 525
Cys Leu Lys Gly Thr Ser Asp Gly Pro Thr Gln Gln Leu Thr Trp Ser
530 535 540
Arg Glu Ser Pro Leu Lys Pro Phe Leu Lys Leu Ser Leu Gly Leu Pro
545 550 555 560
Gly Leu Gly Ile His Met Arg Pro Leu Ala Ile Trp Leu Phe Ile Phe
565 570 575
Asn Val Ser Gln Gln Met Gly Gly Phe Tyr Leu Cys Gln Pro Gly Pro
580 585 590
Pro Ser Glu Lys Ala Trp Gln Pro Gly Trp Thr Val Asn Val Glu Gly
595 600 605
Ser Gly Glu Leu Phe Arg Trp Asn Val Ser Asp Leu Gly Gly Leu Gly
610 615 620
Cys Gly Leu Lys Asn Arg Ser Ser Glu Gly Pro Ser Ser Pro Ser Gly
625 630 635 640
Lys Leu Met Ser Pro Lys Leu Tyr Val Trp Ala Lys Asp Arg Pro Glu
645 650 655
Ile Trp Glu Gly Glu Pro Pro Cys Val Pro Pro Arg Asp Ser Leu Asn
660 665 670
Gln Ser Leu Ser Gln Asp Leu Thr Met Ala Pro Gly Ser Thr Leu Trp
675 680 685
Leu Ser Cys Gly Val Pro Pro Asp Ser Val Ser Arg Gly Pro Leu Ser
690 695 700
Trp Thr His Val His Pro Lys Gly Pro Lys Ser Leu Leu Ser Leu Glu
705 710 715 720
Leu Lys Asp Asp Arg Pro Ala Arg Asp Met Trp Val Met Glu Thr Gly
725 730 735
Leu Leu Leu Pro Arg Ala Thr Ala Gln Asp Ala Gly Lys Tyr Tyr Cys
740 745 750
His Arg Gly Asn Leu Thr Met Ser Phe His Leu Glu Ile Thr Ala Arg
755 760 765
Pro Val Leu Trp His Trp Leu Leu Arg Thr Gly Gly Trp Lys Val Ser
770 775 780
Ala Val Thr Leu Ala Tyr Leu Ile Phe Cys Leu Cys Ser Leu Val Gly
785 790 795 800
Ile Leu His Leu Gln Arg Ala Leu Val Leu Arg Arg Lys Arg
805 810
<210> 30
<211> 921
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-IgG4(HL-CH3)-CD8tm-41BB-Zeta-T2A-CD19t
<400> 30
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
1 5 10 15
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Asp Val Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
His Tyr Thr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
115 120 125
Lys Gly Ser Thr Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
145 150 155 160
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile
165 170 175
Lys Asp Thr Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
180 185 190
Glu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala
195 200 205
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn
210 215 220
Thr Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
225 230 235 240
Tyr Tyr Cys Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Ser Lys Tyr Gly
260 265 270
Pro Pro Cys Pro Pro Cys Pro Gly Gly Gly Ser Ser Gly Gly Gly Ser
275 280 285
Gly Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
290 295 300
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
305 310 315 320
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
325 330 335
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
340 345 350
Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu
355 360 365
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
370 375 380
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Ile Tyr Ile Trp
385 390 395 400
Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile
405 410 415
Thr Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe
420 425 430
Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg
435 440 445
Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Gly Gly Gly Arg Val
450 455 460
Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn
465 470 475 480
Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val
485 490 495
Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg
500 505 510
Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys
515 520 525
Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg
530 535 540
Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys
545 550 555 560
Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg Leu Glu
565 570 575
Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu
580 585 590
Glu Asn Pro Gly Pro Arg Met Pro Pro Pro Arg Leu Leu Phe Phe Leu
595 600 605
Leu Phe Leu Thr Pro Met Glu Val Arg Pro Glu Glu Pro Leu Val Val
610 615 620
Lys Val Glu Glu Gly Asp Asn Ala Val Leu Gln Cys Leu Lys Gly Thr
625 630 635 640
Ser Asp Gly Pro Thr Gln Gln Leu Thr Trp Ser Arg Glu Ser Pro Leu
645 650 655
Lys Pro Phe Leu Lys Leu Ser Leu Gly Leu Pro Gly Leu Gly Ile His
660 665 670
Met Arg Pro Leu Ala Ile Trp Leu Phe Ile Phe Asn Val Ser Gln Gln
675 680 685
Met Gly Gly Phe Tyr Leu Cys Gln Pro Gly Pro Pro Ser Glu Lys Ala
690 695 700
Trp Gln Pro Gly Trp Thr Val Asn Val Glu Gly Ser Gly Glu Leu Phe
705 710 715 720
Arg Trp Asn Val Ser Asp Leu Gly Gly Leu Gly Cys Gly Leu Lys Asn
725 730 735
Arg Ser Ser Glu Gly Pro Ser Ser Pro Ser Gly Lys Leu Met Ser Pro
740 745 750
Lys Leu Tyr Val Trp Ala Lys Asp Arg Pro Glu Ile Trp Glu Gly Glu
755 760 765
Pro Pro Cys Val Pro Pro Arg Asp Ser Leu Asn Gln Ser Leu Ser Gln
770 775 780
Asp Leu Thr Met Ala Pro Gly Ser Thr Leu Trp Leu Ser Cys Gly Val
785 790 795 800
Pro Pro Asp Ser Val Ser Arg Gly Pro Leu Ser Trp Thr His Val His
805 810 815
Pro Lys Gly Pro Lys Ser Leu Leu Ser Leu Glu Leu Lys Asp Asp Arg
820 825 830
Pro Ala Arg Asp Met Trp Val Met Glu Thr Gly Leu Leu Leu Pro Arg
835 840 845
Ala Thr Ala Gln Asp Ala Gly Lys Tyr Tyr Cys His Arg Gly Asn Leu
850 855 860
Thr Met Ser Phe His Leu Glu Ile Thr Ala Arg Pro Val Leu Trp His
865 870 875 880
Trp Leu Leu Arg Thr Gly Gly Trp Lys Val Ser Ala Val Thr Leu Ala
885 890 895
Tyr Leu Ile Phe Cys Leu Cys Ser Leu Val Gly Ile Leu His Leu Gln
900 905 910
Arg Ala Leu Val Leu Arg Arg Lys Arg
915 920
<210> 31
<211> 927
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-IgG4(HL-CH3)-CD28tm-CD28gg-Zeta-T2A-CD19t
<400> 31
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
1 5 10 15
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Asp Val Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
His Tyr Thr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
115 120 125
Lys Gly Ser Thr Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
145 150 155 160
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile
165 170 175
Lys Asp Thr Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
180 185 190
Glu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala
195 200 205
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn
210 215 220
Thr Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
225 230 235 240
Tyr Tyr Cys Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Ser Lys Tyr Gly
260 265 270
Pro Pro Cys Pro Pro Cys Pro Gly Gly Gly Ser Ser Gly Gly Gly Ser
275 280 285
Gly Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
290 295 300
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
305 310 315 320
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
325 330 335
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
340 345 350
Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu
355 360 365
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
370 375 380
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Met Phe Trp Val
385 390 395 400
Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr
405 410 415
Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Gly Gly
420 425 430
His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg
435 440 445
Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg
450 455 460
Ser Gly Gly Gly Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala
465 470 475 480
Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg
485 490 495
Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu
500 505 510
Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn
515 520 525
Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met
530 535 540
Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly
545 550 555 560
Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala
565 570 575
Leu Pro Pro Arg Leu Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu
580 585 590
Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Arg Met Pro Pro Pro
595 600 605
Arg Leu Leu Phe Phe Leu Leu Phe Leu Thr Pro Met Glu Val Arg Pro
610 615 620
Glu Glu Pro Leu Val Val Lys Val Glu Glu Gly Asp Asn Ala Val Leu
625 630 635 640
Gln Cys Leu Lys Gly Thr Ser Asp Gly Pro Thr Gln Gln Leu Thr Trp
645 650 655
Ser Arg Glu Ser Pro Leu Lys Pro Phe Leu Lys Leu Ser Leu Gly Leu
660 665 670
Pro Gly Leu Gly Ile His Met Arg Pro Leu Ala Ile Trp Leu Phe Ile
675 680 685
Phe Asn Val Ser Gln Gln Met Gly Gly Phe Tyr Leu Cys Gln Pro Gly
690 695 700
Pro Pro Ser Glu Lys Ala Trp Gln Pro Gly Trp Thr Val Asn Val Glu
705 710 715 720
Gly Ser Gly Glu Leu Phe Arg Trp Asn Val Ser Asp Leu Gly Gly Leu
725 730 735
Gly Cys Gly Leu Lys Asn Arg Ser Ser Glu Gly Pro Ser Ser Pro Ser
740 745 750
Gly Lys Leu Met Ser Pro Lys Leu Tyr Val Trp Ala Lys Asp Arg Pro
755 760 765
Glu Ile Trp Glu Gly Glu Pro Pro Cys Val Pro Pro Arg Asp Ser Leu
770 775 780
Asn Gln Ser Leu Ser Gln Asp Leu Thr Met Ala Pro Gly Ser Thr Leu
785 790 795 800
Trp Leu Ser Cys Gly Val Pro Pro Asp Ser Val Ser Arg Gly Pro Leu
805 810 815
Ser Trp Thr His Val His Pro Lys Gly Pro Lys Ser Leu Leu Ser Leu
820 825 830
Glu Leu Lys Asp Asp Arg Pro Ala Arg Asp Met Trp Val Met Glu Thr
835 840 845
Gly Leu Leu Leu Pro Arg Ala Thr Ala Gln Asp Ala Gly Lys Tyr Tyr
850 855 860
Cys His Arg Gly Asn Leu Thr Met Ser Phe His Leu Glu Ile Thr Ala
865 870 875 880
Arg Pro Val Leu Trp His Trp Leu Leu Arg Thr Gly Gly Trp Lys Val
885 890 895
Ser Ala Val Thr Leu Ala Tyr Leu Ile Phe Cys Leu Cys Ser Leu Val
900 905 910
Gly Ile Leu His Leu Gln Arg Ala Leu Val Leu Arg Arg Lys Arg
915 920 925
<210> 32
<211> 808
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-接头-CD28tm-CD28gg-Zeta-T2A-CD19t
<400> 32
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
1 5 10 15
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Asp Val Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
His Tyr Thr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
115 120 125
Lys Gly Ser Thr Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
145 150 155 160
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile
165 170 175
Lys Asp Thr Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
180 185 190
Glu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala
195 200 205
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn
210 215 220
Thr Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
225 230 235 240
Tyr Tyr Cys Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Ser Ser
260 265 270
Gly Gly Gly Ser Gly Met Phe Trp Val Leu Val Val Val Gly Gly Val
275 280 285
Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp
290 295 300
Val Arg Ser Lys Arg Ser Arg Gly Gly His Ser Asp Tyr Met Asn Met
305 310 315 320
Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala
325 330 335
Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Gly Gly Gly Arg Val Lys
340 345 350
Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln
355 360 365
Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu
370 375 380
Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg
385 390 395 400
Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met
405 410 415
Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly
420 425 430
Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp
435 440 445
Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg Leu Glu Gly
450 455 460
Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu
465 470 475 480
Asn Pro Gly Pro Arg Met Pro Pro Pro Arg Leu Leu Phe Phe Leu Leu
485 490 495
Phe Leu Thr Pro Met Glu Val Arg Pro Glu Glu Pro Leu Val Val Lys
500 505 510
Val Glu Glu Gly Asp Asn Ala Val Leu Gln Cys Leu Lys Gly Thr Ser
515 520 525
Asp Gly Pro Thr Gln Gln Leu Thr Trp Ser Arg Glu Ser Pro Leu Lys
530 535 540
Pro Phe Leu Lys Leu Ser Leu Gly Leu Pro Gly Leu Gly Ile His Met
545 550 555 560
Arg Pro Leu Ala Ile Trp Leu Phe Ile Phe Asn Val Ser Gln Gln Met
565 570 575
Gly Gly Phe Tyr Leu Cys Gln Pro Gly Pro Pro Ser Glu Lys Ala Trp
580 585 590
Gln Pro Gly Trp Thr Val Asn Val Glu Gly Ser Gly Glu Leu Phe Arg
595 600 605
Trp Asn Val Ser Asp Leu Gly Gly Leu Gly Cys Gly Leu Lys Asn Arg
610 615 620
Ser Ser Glu Gly Pro Ser Ser Pro Ser Gly Lys Leu Met Ser Pro Lys
625 630 635 640
Leu Tyr Val Trp Ala Lys Asp Arg Pro Glu Ile Trp Glu Gly Glu Pro
645 650 655
Pro Cys Val Pro Pro Arg Asp Ser Leu Asn Gln Ser Leu Ser Gln Asp
660 665 670
Leu Thr Met Ala Pro Gly Ser Thr Leu Trp Leu Ser Cys Gly Val Pro
675 680 685
Pro Asp Ser Val Ser Arg Gly Pro Leu Ser Trp Thr His Val His Pro
690 695 700
Lys Gly Pro Lys Ser Leu Leu Ser Leu Glu Leu Lys Asp Asp Arg Pro
705 710 715 720
Ala Arg Asp Met Trp Val Met Glu Thr Gly Leu Leu Leu Pro Arg Ala
725 730 735
Thr Ala Gln Asp Ala Gly Lys Tyr Tyr Cys His Arg Gly Asn Leu Thr
740 745 750
Met Ser Phe His Leu Glu Ile Thr Ala Arg Pro Val Leu Trp His Trp
755 760 765
Leu Leu Arg Thr Gly Gly Trp Lys Val Ser Ala Val Thr Leu Ala Tyr
770 775 780
Leu Ile Phe Cys Leu Cys Ser Leu Val Gly Ile Leu His Leu Gln Arg
785 790 795 800
Ala Leu Val Leu Arg Arg Lys Arg
805
<210> 33
<211> 802
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-接头-CD8tm-41BB-Zeta-T2A-CD19t
<400> 33
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
1 5 10 15
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Asp Val Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
His Tyr Thr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
115 120 125
Lys Gly Ser Thr Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
145 150 155 160
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile
165 170 175
Lys Asp Thr Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
180 185 190
Glu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala
195 200 205
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn
210 215 220
Thr Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
225 230 235 240
Tyr Tyr Cys Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Ser Ser
260 265 270
Gly Gly Gly Ser Gly Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys
275 280 285
Gly Val Leu Leu Leu Ser Leu Val Ile Thr Lys Arg Gly Arg Lys Lys
290 295 300
Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr
305 310 315 320
Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly
325 330 335
Gly Cys Glu Leu Gly Gly Gly Arg Val Lys Phe Ser Arg Ser Ala Asp
340 345 350
Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn
355 360 365
Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg
370 375 380
Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly
385 390 395 400
Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu
405 410 415
Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu
420 425 430
Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His
435 440 445
Met Gln Ala Leu Pro Pro Arg Leu Glu Gly Gly Gly Glu Gly Arg Gly
450 455 460
Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Arg Met
465 470 475 480
Pro Pro Pro Arg Leu Leu Phe Phe Leu Leu Phe Leu Thr Pro Met Glu
485 490 495
Val Arg Pro Glu Glu Pro Leu Val Val Lys Val Glu Glu Gly Asp Asn
500 505 510
Ala Val Leu Gln Cys Leu Lys Gly Thr Ser Asp Gly Pro Thr Gln Gln
515 520 525
Leu Thr Trp Ser Arg Glu Ser Pro Leu Lys Pro Phe Leu Lys Leu Ser
530 535 540
Leu Gly Leu Pro Gly Leu Gly Ile His Met Arg Pro Leu Ala Ile Trp
545 550 555 560
Leu Phe Ile Phe Asn Val Ser Gln Gln Met Gly Gly Phe Tyr Leu Cys
565 570 575
Gln Pro Gly Pro Pro Ser Glu Lys Ala Trp Gln Pro Gly Trp Thr Val
580 585 590
Asn Val Glu Gly Ser Gly Glu Leu Phe Arg Trp Asn Val Ser Asp Leu
595 600 605
Gly Gly Leu Gly Cys Gly Leu Lys Asn Arg Ser Ser Glu Gly Pro Ser
610 615 620
Ser Pro Ser Gly Lys Leu Met Ser Pro Lys Leu Tyr Val Trp Ala Lys
625 630 635 640
Asp Arg Pro Glu Ile Trp Glu Gly Glu Pro Pro Cys Val Pro Pro Arg
645 650 655
Asp Ser Leu Asn Gln Ser Leu Ser Gln Asp Leu Thr Met Ala Pro Gly
660 665 670
Ser Thr Leu Trp Leu Ser Cys Gly Val Pro Pro Asp Ser Val Ser Arg
675 680 685
Gly Pro Leu Ser Trp Thr His Val His Pro Lys Gly Pro Lys Ser Leu
690 695 700
Leu Ser Leu Glu Leu Lys Asp Asp Arg Pro Ala Arg Asp Met Trp Val
705 710 715 720
Met Glu Thr Gly Leu Leu Leu Pro Arg Ala Thr Ala Gln Asp Ala Gly
725 730 735
Lys Tyr Tyr Cys His Arg Gly Asn Leu Thr Met Ser Phe His Leu Glu
740 745 750
Ile Thr Ala Arg Pro Val Leu Trp His Trp Leu Leu Arg Thr Gly Gly
755 760 765
Trp Lys Val Ser Ala Val Thr Leu Ala Tyr Leu Ile Phe Cys Leu Cys
770 775 780
Ser Leu Val Gly Ile Leu His Leu Gln Arg Ala Leu Val Leu Arg Arg
785 790 795 800
Lys Arg
<210> 34
<211> 658
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-IgG4(L235E,
N297Q)-CD28tm-CD28gg-Zeta-T2A-CD19t 减去引导物, TSA或CD19t
<400> 34
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Glu Val Gln
115 120 125
Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
130 135 140
Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His
145 150 155 160
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile
165 170 175
Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg
180 185 190
Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met
195 200 205
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp
210 215 220
Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu
225 230 235 240
Val Thr Val Ser Ser Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys
245 250 255
Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
260 265 270
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
275 280 285
Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp
290 295 300
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
305 310 315 320
Glu Gln Phe Gln Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
325 330 335
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
340 345 350
Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
355 360 365
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu
370 375 380
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
385 390 395 400
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
405 410 415
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
420 425 430
Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn
435 440 445
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
450 455 460
Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Met Phe Trp Val Leu Val
465 470 475 480
Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala
485 490 495
Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Gly Gly His Ser
500 505 510
Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His
515 520 525
Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Gly
530 535 540
Gly Gly Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln
545 550 555 560
Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu
565 570 575
Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly
580 585 590
Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu
595 600 605
Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly
610 615 620
Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser
625 630 635 640
Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro
645 650 655
Pro Arg
<210> 35
<211> 652
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合
Her2scFv-IgG4(L235E,N297Q)-CD8tm-41BB-Zeta-T2A-CD19t减去
引导物, TSA或CD19t
<400> 35
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Glu Val Gln
115 120 125
Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
130 135 140
Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His
145 150 155 160
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile
165 170 175
Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg
180 185 190
Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met
195 200 205
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp
210 215 220
Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu
225 230 235 240
Val Thr Val Ser Ser Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys
245 250 255
Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
260 265 270
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
275 280 285
Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp
290 295 300
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
305 310 315 320
Glu Gln Phe Gln Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
325 330 335
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
340 345 350
Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
355 360 365
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu
370 375 380
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
385 390 395 400
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
405 410 415
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
420 425 430
Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn
435 440 445
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
450 455 460
Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Ile Tyr Ile Trp Ala Pro
465 470 475 480
Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Lys
485 490 495
Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg
500 505 510
Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro
515 520 525
Glu Glu Glu Glu Gly Gly Cys Glu Leu Gly Gly Gly Arg Val Lys Phe
530 535 540
Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu
545 550 555 560
Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp
565 570 575
Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys
580 585 590
Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala
595 600 605
Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys
610 615 620
Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr
625 630 635 640
Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
645 650
<210> 36
<211> 474
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-CD8铰链-CD8tm-41BB-Zeta-T2A-CD19t 减去
引导物, TSA或CD19t
<400> 36
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Glu Val Gln
115 120 125
Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
130 135 140
Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His
145 150 155 160
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile
165 170 175
Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg
180 185 190
Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met
195 200 205
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp
210 215 220
Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu
225 230 235 240
Val Thr Val Ser Ser Ala Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro
245 250 255
Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro
260 265 270
Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu
275 280 285
Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys
290 295 300
Gly Val Leu Leu Leu Ser Leu Val Ile Thr Gly Gly Gly Lys Arg Gly
305 310 315 320
Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val
325 330 335
Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu
340 345 350
Glu Glu Gly Gly Cys Glu Leu Gly Gly Gly Arg Val Lys Phe Ser Arg
355 360 365
Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn
370 375 380
Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg
385 390 395 400
Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro
405 410 415
Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala
420 425 430
Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His
435 440 445
Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp
450 455 460
Ala Leu His Met Gln Ala Leu Pro Pro Arg
465 470
<210> 37
<211> 445
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-铰链-接头-CD8tm-41BB-Zeta-T2A-CD19t
减去引导物, TSA或CD19t
<400> 37
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Glu Val Gln
115 120 125
Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
130 135 140
Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His
145 150 155 160
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile
165 170 175
Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg
180 185 190
Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met
195 200 205
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp
210 215 220
Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu
225 230 235 240
Val Thr Val Ser Ser Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys
245 250 255
Pro Gly Gly Gly Ser Ser Gly Gly Gly Ser Gly Ile Tyr Ile Trp Ala
260 265 270
Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr
275 280 285
Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met
290 295 300
Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe
305 310 315 320
Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Gly Gly Gly Arg Val Lys
325 330 335
Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln
340 345 350
Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu
355 360 365
Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg
370 375 380
Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met
385 390 395 400
Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly
405 410 415
Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp
420 425 430
Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
435 440 445
<210> 38
<211> 552
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-IgG4(HL-CH3)-CD8tm-41BB-Zeta-T2A-CD19t
减去引导物, TSA或CD19t
<400> 38
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Glu Val Gln
115 120 125
Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
130 135 140
Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His
145 150 155 160
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile
165 170 175
Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg
180 185 190
Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met
195 200 205
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp
210 215 220
Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu
225 230 235 240
Val Thr Val Ser Ser Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys
245 250 255
Pro Gly Gly Gly Ser Ser Gly Gly Gly Ser Gly Gly Gln Pro Arg Glu
260 265 270
Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn
275 280 285
Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
290 295 300
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
305 310 315 320
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg
325 330 335
Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys
340 345 350
Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
355 360 365
Ser Leu Ser Leu Gly Lys Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr
370 375 380
Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Lys Arg Gly Arg Lys
385 390 395 400
Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr
405 410 415
Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu
420 425 430
Gly Gly Cys Glu Leu Gly Gly Gly Arg Val Lys Phe Ser Arg Ser Ala
435 440 445
Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu
450 455 460
Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly
465 470 475 480
Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu
485 490 495
Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser
500 505 510
Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly
515 520 525
Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu
530 535 540
His Met Gln Ala Leu Pro Pro Arg
545 550
<210> 39
<211> 558
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-IgG4(HL-CH3)-CD28tm-CD28gg-Zeta-T2A-CD19t
减去引导物, TSA或CD19t
<400> 39
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Glu Val Gln
115 120 125
Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
130 135 140
Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His
145 150 155 160
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile
165 170 175
Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg
180 185 190
Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met
195 200 205
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp
210 215 220
Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu
225 230 235 240
Val Thr Val Ser Ser Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys
245 250 255
Pro Gly Gly Gly Ser Ser Gly Gly Gly Ser Gly Gly Gln Pro Arg Glu
260 265 270
Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn
275 280 285
Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
290 295 300
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
305 310 315 320
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg
325 330 335
Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys
340 345 350
Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
355 360 365
Ser Leu Ser Leu Gly Lys Met Phe Trp Val Leu Val Val Val Gly Gly
370 375 380
Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe
385 390 395 400
Trp Val Arg Ser Lys Arg Ser Arg Gly Gly His Ser Asp Tyr Met Asn
405 410 415
Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr
420 425 430
Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Gly Gly Gly Arg Val
435 440 445
Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn
450 455 460
Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val
465 470 475 480
Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg
485 490 495
Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys
500 505 510
Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg
515 520 525
Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys
530 535 540
Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
545 550 555
<210> 40
<211> 439
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-接头-CD28tm-CD28gg-Zeta-T2A-CD19t 减去
引导物, TSA或CD19t
<400> 40
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Glu Val Gln
115 120 125
Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
130 135 140
Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His
145 150 155 160
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile
165 170 175
Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg
180 185 190
Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met
195 200 205
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp
210 215 220
Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu
225 230 235 240
Val Thr Val Ser Ser Gly Gly Gly Ser Ser Gly Gly Gly Ser Gly Met
245 250 255
Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu
260 265 270
Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser
275 280 285
Arg Gly Gly His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly
290 295 300
Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala
305 310 315 320
Ala Tyr Arg Ser Gly Gly Gly Arg Val Lys Phe Ser Arg Ser Ala Asp
325 330 335
Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn
340 345 350
Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg
355 360 365
Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly
370 375 380
Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu
385 390 395 400
Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu
405 410 415
Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His
420 425 430
Met Gln Ala Leu Pro Pro Arg
435
<210> 41
<211> 433
<212> PRT
<213> 人工序列
<220>
<223> 人的 嵌合Her2scFv-接头-CD8tm-41BB-Zeta-T2A-CD19t 减去
引导物, TSA或CD19t
<400> 41
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Glu Val Gln
115 120 125
Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
130 135 140
Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His
145 150 155 160
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile
165 170 175
Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg
180 185 190
Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met
195 200 205
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp
210 215 220
Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu
225 230 235 240
Val Thr Val Ser Ser Gly Gly Gly Ser Ser Gly Gly Gly Ser Gly Ile
245 250 255
Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser
260 265 270
Leu Val Ile Thr Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys
275 280 285
Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys
290 295 300
Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Gly Gly
305 310 315 320
Gly Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln
325 330 335
Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu
340 345 350
Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly
355 360 365
Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln
370 375 380
Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu
385 390 395 400
Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr
405 410 415
Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro
420 425 430
Arg
<210> 42
<211> 22
<212> PRT
<213> 人工序列
<220>
<223> 铰链-接头域
<400> 42
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Gly Gly Gly Ser
1 5 10 15
Ser Gly Gly Gly Ser Gly
20

Claims (16)

1.编码嵌合抗原受体的核酸分子，其中所述核酸分子表达选自SEQ ID NO: 26和SEQID NO: 27的氨基酸序列的多肽。
2.权利要求1的核酸分子，其中所述核酸分子表达SEQ ID NO: 26的多肽。
3.权利要求1的核酸分子，其中所述核酸分子表达SEQ ID NO: 27的多肽。
4.人T细胞的群体，所述人T细胞包含核酸，所述核酸表达选自SEQ ID NO: 26 和27的氨基酸序列的嵌合抗原受体。
5.人T细胞的群体，所述人T细胞包含载体，所述载体表达选自SEQ ID NO: 26 和27的氨基酸序列的嵌合抗原受体。
6.人T细胞群体，所述人T细胞通过载体转导，所述载体包含编码选自SEQ ID NO: 26和SEQ ID NO: 27的氨基酸序列的嵌合抗原受体的表达盒。
7.权利要求6的人T细胞群体，其中所述T细胞包含中央记忆T细胞群体。
8.通过包含编码嵌合抗原受体的表达盒的载体转导的自体或同种异体人T细胞群体，所述自体或同种异体人T细胞群体用于治疗患者中的表达HER2的癌症，其中对所述患者施用自体或同种异体人T细胞群体，并且其中嵌合抗原受体是选自SEQ ID NO: 26和27的氨基酸序列。
9.权利要求8的自体或同种异体人T细胞群体，其中所述人T细胞群体包含CD62L+记忆T细胞。
10.权利要求8的自体或同种异体人T细胞群体，其中所述癌症是乳腺癌、脑癌、或乳房至脑的转移。
11.权利要求8的自体或同种异体人T细胞群体，其中所述转导的人T细胞通过方法制备，所述方法包括从所述患者获得T细胞，处理所述T细胞以分离中央记忆T细胞并且用病毒载体转导所述中央记忆细胞的至少部分，所述病毒载体包含编码嵌合抗原受体的表达盒，其中嵌合抗原受体是选自SEQ ID NO: 26和27的氨基酸序列。
12.权利要求8的自体或同种异体人T细胞群体，其中肿瘤内或心室内施用所述T细胞。
13.表达多肽的T细胞，所述多肽的氨基酸序列选自SEQ ID NO: 26和27。
14.权利要求13的T细胞，其中所述氨基酸序列是SEQ ID NO:26。
15.权利要求13的T细胞，其中所述氨基酸序列是SEQ ID NO:27。
16.多肽，其选自SEQ ID NO: 26和27。
CN201680076412.0A 2015-11-04 2016-11-04 靶向her2的嵌合抗原受体 Active CN108779174B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202211409883.2A CN116063574A (zh) 2015-11-04 2016-11-04 靶向her2的嵌合抗原受体

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201562251052P 2015-11-04 2015-11-04
US62/251,052 2015-11-04
PCT/US2016/060724 WO2017079694A2 (en) 2015-11-04 2016-11-04 Chimeric antigen receptors targeting her2

Related Child Applications (1)

Application Number Title Priority Date Filing Date
CN202211409883.2A Division CN116063574A (zh) 2015-11-04 2016-11-04 靶向her2的嵌合抗原受体

Publications (2)

Publication Number Publication Date
CN108779174A CN108779174A (zh) 2018-11-09
CN108779174B true CN108779174B (zh) 2022-11-29

Family

ID=58662864

Family Applications (2)

Application Number Title Priority Date Filing Date
CN201680076412.0A Active CN108779174B (zh) 2015-11-04 2016-11-04 靶向her2的嵌合抗原受体
CN202211409883.2A Pending CN116063574A (zh) 2015-11-04 2016-11-04 靶向her2的嵌合抗原受体

Family Applications After (1)

Application Number Title Priority Date Filing Date
CN202211409883.2A Pending CN116063574A (zh) 2015-11-04 2016-11-04 靶向her2的嵌合抗原受体

Country Status (13)

Country Link
US (2) US11197919B2 (zh)
EP (2) EP3371227B8 (zh)
JP (3) JP6932709B2 (zh)
KR (1) KR20180083874A (zh)
CN (2) CN108779174B (zh)
AU (2) AU2016349722A1 (zh)
BR (1) BR112018009129A2 (zh)
CA (1) CA3004306A1 (zh)
ES (1) ES2909973T3 (zh)
IL (2) IL259120B (zh)
MX (2) MX2018005674A (zh)
RU (1) RU2753695C2 (zh)
WO (1) WO2017079694A2 (zh)

Families Citing this family (29)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106459914B (zh) 2014-05-15 2020-11-06 新加坡国立大学 经修饰的自然杀伤细胞及其用途
WO2017079694A2 (en) * 2015-11-04 2017-05-11 Priceman Saul J Chimeric antigen receptors targeting her2
US20170224733A1 (en) * 2016-02-05 2017-08-10 City Of Hope Administration of Engineered T Cells for Treatment of Cancers in the Central Nervous System
SG11201907299XA (en) 2017-02-08 2019-09-27 Dragonfly Therapeutics Inc Multi-specific binding proteins for activation of natural killer cells and therapeutic uses thereof to treat cancer
ES2955074T3 (es) 2017-02-20 2023-11-28 Dragonfly Therapeutics Inc Proteínas que se unen a HER2, NKG2D Y CD16
US11896616B2 (en) 2017-03-27 2024-02-13 National University Of Singapore Stimulatory cell lines for ex vivo expansion and activation of natural killer cells
MX2019011514A (es) 2017-03-27 2020-01-27 Nat Univ Singapore Receptores quimericos nkg2d truncados y usos de los mismos en inmunoterapia con celulas asesinas naturales.
US20210179733A1 (en) * 2017-11-14 2021-06-17 Green Cross Lab Cell Corporation Anti-her2 antibody or antigen-binding fragment thereof, and chimeric antigen receptor comprising same
US11649294B2 (en) * 2017-11-14 2023-05-16 GC Cell Corporation Anti-HER2 antibody or antigen-binding fragment thereof, and chimeric antigen receptor comprising same
CN109608547B (zh) * 2017-12-29 2022-03-15 郑州大学第一附属医院 表达Her2的嵌合抗原受体、慢病毒表达载体及其应用
WO2019157366A1 (en) 2018-02-08 2019-08-15 Dragonfly Therapeutics, Inc. Antibody variable domains targeting the nkg2d receptor
US20210054409A1 (en) * 2018-02-09 2021-02-25 National University Of Singapore Adhesion Receptor Constructs and Uses Thereof in Natural Killer Cell Immunotherapy
CN111954684B (zh) * 2018-02-11 2024-05-07 加利福尼亚大学董事会 Car-t细胞与自身免疫性疾病
SG10201801219VA (en) * 2018-02-13 2019-09-27 Agency Science Tech & Res Anti-HER2 Antibodies
US20210252058A1 (en) * 2018-05-17 2021-08-19 St. Jude Children's Research Hospital, Inc. Chimeric antigen receptors with myd88 and cd40 costimulatory domains
EA202091888A1 (ru) * 2018-08-08 2020-10-23 Драгонфлай Терапьютикс, Инк. Вариабельные домены антител, нацеленные на рецептор nkg2d
US20230065784A1 (en) 2019-03-21 2023-03-02 City Of Hope Composition and method for reducing expression of checkpoint inhibitors in t cells expressing a car or ctl
AU2021210446A1 (en) * 2020-01-23 2022-07-28 Bioatla, Inc. Chimeric antigen receptors to HER2 and methods of use thereof
CA3184449A1 (en) * 2020-05-22 2021-11-25 GC Cell Corporation Anti-her2 antibody or antigen-binding fragment thereof, and chimeric antigen receptor comprising same
CN112481284B (zh) * 2020-12-07 2023-07-25 深圳瑞吉生物科技有限公司 一种编码CAR基因的mRNA、组合mRNA、构建方法、CAR-T细胞和应用
WO2022187182A1 (en) * 2021-03-02 2022-09-09 The Trustees Of The University Of Pennsylvania Targeting t regulatory cells to islet cells to stall or reverse type 1 diabetes
CN117242090A (zh) 2021-03-30 2023-12-15 希望之城公司 CAR T细胞疗法和IFNγ
WO2023274385A1 (zh) * 2021-07-01 2023-01-05 宁波茂行生物医药科技有限公司 靶向her2的通用型car-t细胞及其制备方法
CA3226798A1 (en) 2021-07-29 2023-02-02 James MATTHAEI Synovial extracellular matrix-specific chimeric antigen receptor for targeting regulatory t cells to treat autoimmune diseases
CN116023491A (zh) 2021-10-25 2023-04-28 江苏众红生物工程创药研究院有限公司 抗cd26抗体及其应用
WO2023172514A1 (en) * 2022-03-07 2023-09-14 Catamaran Bio, Inc. Engineered immune cell therapeutics targeted to her2 and methods of use thereof
WO2023173116A1 (en) 2022-03-10 2023-09-14 City Of Hope Membrane-bound il-12 for cellular immunotherapy
JP2024515920A (ja) * 2022-04-08 2024-04-11 フェイト セラピューティクス，インコーポレイティド 固形腫瘍標的化骨格を有する細胞及びその使用
CN114805584B (zh) * 2022-06-30 2022-09-09 上海优替济生生物医药有限公司 抗原结合蛋白及其用途

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011137245A2 (en) * 2010-04-30 2011-11-03 Esperance Pharmaceuticals, Inc. Lytic-peptide-her2/neu (human epidermal growth factor receptor 2) ligand conjugates and methods of use
WO2014100385A1 (en) * 2012-12-20 2014-06-26 Anthrogenesis Corporation Chimeric antigen receptors

Family Cites Families (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1918303A3 (en) * 1999-04-09 2008-05-14 Universität Zürich method for the stabilization of chimeric immunoglobulins or immunoglobulin fragments, and stabilized anti-egp-2 scfv fragment
JP2008507520A (ja) 2004-07-22 2008-03-13 ジェネンテック・インコーポレーテッド Ｈｅｒ２抗体組成物
BRPI0811857A2 (pt) * 2007-05-14 2014-10-21 Biogen Idec Inc Regiões fc (scfc) de cadeia simples, polipeptídeos de aglutinação que as compreendem e métodos relacionados.
CA2779526C (en) 2009-11-03 2022-12-06 City Of Hope Truncated epiderimal growth factor receptor (egfrt) for transduced t cell selection
US8491908B2 (en) * 2010-06-01 2013-07-23 Canon Kabushiki Kaisha Composite particle, contrast agent for photoacoustic imaging, and method for producing the composite particle
WO2012050374A2 (en) 2010-10-13 2012-04-19 Innocell, Inc. Immunotherapy for solid tumors
AU2012230780B2 (en) 2011-03-23 2016-10-27 Fred Hutchinson Cancer Center Method and compositions for cellular immunotherapy
PL2718322T3 (pl) * 2011-06-06 2019-07-31 Novo Nordisk A/S Lecznicze przeciwciała
RS61345B1 (sr) * 2012-08-20 2021-02-26 Hutchinson Fred Cancer Res Postupak i kompozicije za ćelijsku imunoterapiju
EP4029937A1 (en) * 2012-10-04 2022-07-20 Research Development Foundation Serine protease molecules and therapies
WO2014055657A1 (en) 2012-10-05 2014-04-10 The Trustees Of The University Of Pennsylvania Use of a trans-signaling approach in chimeric antigen receptors
DK2914633T3 (da) * 2012-10-30 2022-03-14 Esperance Pharmaceuticals Inc Antistof/lægemiddel-konjugater og anvendelsesfremgangsmåder
US9657105B2 (en) * 2013-03-15 2017-05-23 City Of Hope CD123-specific chimeric antigen receptor redirected T cells and methods of their use
EP3039040B1 (en) * 2013-08-26 2021-12-22 Hinrich Abken Anti cd30 chimeric antigen receptor and its use
CA2936501A1 (en) * 2014-01-13 2015-07-16 Stephen J. Forman Chimeric antigen receptors (cars) having mutations in the fc spacer region and methods for their use
CA2945388A1 (en) * 2014-04-23 2015-10-29 Board Of Regents, The University Of Texas System Chimeric antigen receptors (car) for use in therapy and methods for making the same
WO2017079694A2 (en) * 2015-11-04 2017-05-11 Priceman Saul J Chimeric antigen receptors targeting her2

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011137245A2 (en) * 2010-04-30 2011-11-03 Esperance Pharmaceuticals, Inc. Lytic-peptide-her2/neu (human epidermal growth factor receptor 2) ligand conjugates and methods of use
WO2014100385A1 (en) * 2012-12-20 2014-06-26 Anthrogenesis Corporation Chimeric antigen receptors

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
A Herceptin-based chimeric antigen receptor with modified signaling domains leads to enhanced survival of transduced T lymphocytes and antitumor activity;Yangbing Zhao et al;《The Journal of Immunology》;20091231;第183卷;5563-5574 *
Chimeric antigen receptors with mutated IgG4 Fc spacer avoid Fc receptor binding and improve T cell persistence and antitumor efficacy;Jonnalagadda M et al;《Molecular Therapy》;20150217;第23卷(第4期);摘要 *
Jonnalagadda M et al.Chimeric antigen receptors with mutated IgG4 Fc spacer avoid Fc receptor binding and improve T cell persistence and antitumor efficacy.《Molecular Therapy》.2015,第23卷(第4期), *
Yangbing Zhao et al.A Herceptin-based chimeric antigen receptor with modified signaling domains leads to enhanced survival of transduced T lymphocytes and antitumor activity.《The Journal of Immunology》.2009,第183卷 *

Also Published As

Publication number Publication date
MX2018005674A (es) 2019-01-10
EP4074731A1 (en) 2022-10-19
MX2022013498A (es) 2022-11-16
CA3004306A1 (en) 2017-05-11
US11197919B2 (en) 2021-12-14
RU2753695C2 (ru) 2021-08-19
IL259120A (en) 2018-06-28
CN108779174A (zh) 2018-11-09
EP3371227A2 (en) 2018-09-12
US20220265796A1 (en) 2022-08-25
JP2021192617A (ja) 2021-12-23
EP3371227B8 (en) 2022-04-06
RU2018118817A3 (zh) 2020-03-11
CN116063574A (zh) 2023-05-05
JP2023100653A (ja) 2023-07-19
JP2019500894A (ja) 2019-01-17
JP7264954B2 (ja) 2023-04-25
IL292222A (en) 2022-06-01
JP6932709B2 (ja) 2021-09-08
IL259120B (en) 2022-05-01
WO2017079694A2 (en) 2017-05-11
AU2022200116A1 (en) 2022-02-10
EP3371227A4 (en) 2018-09-12
ES2909973T3 (es) 2022-05-10
US20180326032A1 (en) 2018-11-15
KR20180083874A (ko) 2018-07-23
EP3371227B1 (en) 2022-02-23
AU2016349722A1 (en) 2018-05-24
BR112018009129A2 (pt) 2019-02-26
RU2018118817A (ru) 2019-12-05
WO2017079694A3 (en) 2017-07-20

Similar Documents

Publication Publication Date Title
CN108779174B (zh) 靶向her2的嵌合抗原受体
AU2021200122B2 (en) CS1 Targeted Chimeric Antigen Receptor-Modified T Cells
JP7297854B2 (ja) Ｐｓｃａを標的とするキメラ抗原受容体
CN107002084B (zh) 靶向IL13Rα2的共刺激嵌合抗原受体T细胞
EP3362470B1 (en) Chimeric antigen receptors containing a chlorotoxin domain

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 40000323

Country of ref document: HK

GR01 Patent grant
GR01 Patent grant