WO2010058023A1 - Récepteur de lymphocytes t de forte affinité et ses applications - Google Patents

Récepteur de lymphocytes t de forte affinité et ses applications Download PDF

Info

Publication number
WO2010058023A1
WO2010058023A1 PCT/EP2009/065705 EP2009065705W WO2010058023A1 WO 2010058023 A1 WO2010058023 A1 WO 2010058023A1 EP 2009065705 W EP2009065705 W EP 2009065705W WO 2010058023 A1 WO2010058023 A1 WO 2010058023A1
Authority
WO
WIPO (PCT)
Prior art keywords
tcr
cells
chain
cell
hla
Prior art date
Application number
PCT/EP2009/065705
Other languages
English (en)
Inventor
Dolores Schendel
Susanne Wilde
Bernhard Frankenberger
Wolfgang Uckert
Original Assignee
Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt Gmbh
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
Priority to CA2743669A priority Critical patent/CA2743669C/fr
Priority to CN200980154272.4A priority patent/CN102272153B/zh
Priority to ES09760145T priority patent/ES2394180T3/es
Priority to AU2009317161A priority patent/AU2009317161B2/en
Priority to US13/130,665 priority patent/US8697854B2/en
Priority to EP09760145A priority patent/EP2352756B1/fr
Application filed by Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt Gmbh filed Critical Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt Gmbh
Priority to JP2011536895A priority patent/JP6069755B2/ja
Priority to DK09760145.4T priority patent/DK2352756T3/da
Publication of WO2010058023A1 publication Critical patent/WO2010058023A1/fr
Priority to US14/224,525 priority patent/US9862755B2/en
Priority to US15/822,970 priority patent/US10626159B2/en
Priority to US16/820,856 priority patent/US20200339653A1/en

Links

Classifications

    • 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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
    • A61K38/1774—Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules)
    • 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
    • A61K39/4632—T-cell receptors [TCR]; antibody T-cell receptor constructs
    • 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
    • A61K39/464454—Enzymes
    • A61K39/464456—Tyrosinase or tyrosinase related proteinases [TRP-1 or TRP-2]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation

Definitions

  • the present invention is directed to a high affinity T cell receptor (TCR) against a tumor- associated antigen, an isolated nucleic acid molecule encoding same, a T cell expressing said TCR, and a pharmaceutical composition for use in the treatment of diseases involving malignant cells expressing said tumor-associated antigen.
  • TCR T cell receptor
  • TCRs are members of the immunoglobulin superfamily and usually consist of two subunits, namely the ⁇ - and ⁇ -subunits. These possess one N-terminal immunoglobulin (Ig)-variable (V) domain, one Ig-constant (C) domain, a transmembrane/cell membrane-spanning region, and a short cytoplasmic tail at the C-terminal end.
  • the variable domains of both the TCR ⁇ - chain and ⁇ -chain have three hypervariable or complementarity determining regions (CDRs), whereas the variable region of the ⁇ -chain has an additional area of hypervariability (HV4) that does not normally contact antigen and therefore is not considered a CDR.
  • CDR3 is the main CDR responsible for recognizing processed antigen, although CDRl of the alpha chain has also been shown to interact with the N-terminal part of the antigenic peptide, whereas CDRl of the ⁇ -chain interacts with the C-terminal part of the peptide.
  • CDR2 is thought to recognize the MHC.
  • CDR4 of the ⁇ -chain is not thought to participate in antigen recognition, but has been shown to interact with superantigens.
  • the constant domain of the TCR domain consists of short connecting sequences in which a cysteine residue forms disulfide bonds, which forms a link between the two chains.
  • the affinity of TCR' s for a specific antigen makes them valuable for several therapeutic approaches. For example, cancer patients, such as melanoma patients, can be effectively treated by using adoptive immunotherapy.
  • SCT allogeneic stem cell transplantation
  • a transfer of unprimed, unseparated lymphocytes can be made to speed up the generation of immune responses directed against tumor cells.
  • the specific lymphocytes capable of attacking tumor cells must be activated and expanded from the low frequency precursor lymphocytes that are present among the unselected population of lymphocytes that are transferred.
  • Donor lymphocyte infusions (DLI) of unselected lymphocyte populations after SCT work well for the elimination of chronic myelogenous leukemia (CML), which grows slowly, but are less effective in the eradication of acute leukemia, partly due to the fact that the growth of the malignant cells outpaces the expansion capacity of the immune cells.
  • CML chronic myelogenous leukemia
  • T cells may also be transferred that have the capacity to attack normal cells and tissues of the recipient, leading to graft-versus-host-disease (GVHD), a disease with high morbidity and mortality.
  • GVHD graft-versus-host-disease
  • TCR T cell receptors
  • an expanded allospecific T cell clone that is specific for an MHC molecule not expressed by the recipient or an expanded T cell clone specific for a virus, such as cytomegalovirus or Epstein-Barr virus, could be used as recipient cells for the transgenic TCR.
  • the availability of a panel of transgenic TCR vectors, recognizing different MHC- peptide ligands could be used to develop large numbers of pre-activated T cells of both the CD4 and CD8 subtypes, thereby allowing large numbers of effector lymphocytes to be rapidly prepared and transferred to patients whose tumors express the corresponding TCR ligands. This would save time in achieving the numbers of specific T cells required to control tumor growth, possibly leading to more effective tumor eradication of rapidly progressing tumors.
  • TCR T cell receptors
  • WO 2006/031221 pertains to T cell receptors against tumor-associated antigens, nucleic acids encoding the same, vectors and cells comprising the nucleic acids encoding the T cell receptors, and methods of use thereof.
  • the TCR subunits have the ability to form TCR that confer specificity to T cells for tumor cells presenting MART-I, NY-ESO-I, and melanoma-related gplOO.
  • Roszkowski et al. J. Immunol. (2003) 170, 2582-2589 and Cancer Res. (2005) 65, 1570- 1576
  • the like are characterising tyrosinase-specific TCR.
  • US 5,906,936 is directed to cytotoxic T-cells which kill non-MHC-restricted target cells and not to cells, which comprise specific TCR sequences.
  • WO97/32603 is directed to a method for producing non-human TCR and TCR specific for human HLA-restricted tumor antigens. Furthermore, the TCR-nucleic acids and recombinant T-cells are described as well as the administration of TCR recombinant T-cells for the treatment of several diseases.
  • WO2007/065957 describes an effector T-cell transfected with an antigen specific TCR coding RNA wherein the transfected T-cell recognizes the antigen in a complex with the MHC-molecule and binds the same.
  • MART-I Melan-A
  • tyrosinase As a potential tumor antigen, MART-I (Melan-A), tyrosinase and survivin are named.
  • WO2008/039818 discloses MART-I and tyrosinase-specific TCR sequences and describes the enhancement of antigen recognition by substitution in the CDR2 region.
  • TCR sequences are all derived from autologous or xenogeneic, but not allogeneic, sources.
  • TCR sequences are from peripheral blood or from tumor infiltrating lymphocytes of HLA- A2 positive melanoma patients. This means that all these TCR are HLA-A2 self-restricted TCRs, or, are HLA-DP4 restricted, NY-ESO-I specific, both derived from autologous sources.
  • the TCR is derived from an HLA- A2 transgenic mouse, so in this case the sequence is xenogeneic.
  • the available prior art documents do not show TCR sequences, which are allo- restricted and tyrosinase-specif ⁇ c.
  • TCR transgenic T cell receptors
  • TCR or functional parts thereof such as CDR3 regions, which show high affinity against tumor-associated antigens, in particular tyrosinase.
  • pharmaceutical compositions for use in adoptive cell therapy which allow an effective treatment of diseases involving malignant cells expressing tyrosinase, preferably melanomas, gliomas, glioblastomas, and/or rare tumors of ectodermal origin.
  • TCR specific for the melanoma-associated antigen, tyrosinase could be isolated by the inventors and it could be shown that TCR derived from the allo-restricted clone were superior in recognition of specific peptide and tumor cells after expression as transgenes in recipient lymphocytes. Therefore, TCRs and functional parts thereof, such as CDR3 regions could be identified, which find application in adoptive cell therapy for the treatment of several malignancies.
  • T cell clones with specificity for various tumor-associated antigens have been reported over the years (see above). Most of these TCR are restricted by self-MHC molecules.
  • the TCR sequences disclosed herein, however, are allo-restricted and show high-avidity in recognition of their specific ligands.
  • the TCR of the present invention are not self-MHC-restricted and therefore have higher structural affinity for interactions with MHC-peptide ligands that target tumor cells via common over-expressed self proteins.
  • the TCR of the present invention were derived from a T cell clone generated by priming CD 8* T cells with autologous dendritic cells from an HLA- A2 negative donor co-expressing allogeneic HLA-*A0201 molecules and an antigen.
  • the present TCR are of therapeutic use for the treatment of HLA- A2 positive patients.
  • T cell responses against tumors are often directed against self-MHC molecules presenting peptides derived from over-expressed self-proteins.
  • T cells with high avidity for self-peptide/self-MHC ligands are eliminated by negative selection to prevent autoimmunity.
  • the TCR affinity of remaining T cells specific for self-ligands is normally low, however high-avidity T cells are needed to effectively eradicate tumors.
  • negative selection is limited to self-MHC molecules, T cells that recognize allogeneic MHC molecules have not undergone negative selection.
  • peptides are presented by allogeneic MHC molecules, it is feasible to obtain high-avidity T cells specific for common tumor-associated ligands derived from over-expressed self-proteins.
  • the present invention provides a nucleic acid molecule coding for the V(D)J regions of a TCR that recognizes a tumor antigen and comprising the nucleic acid sequence of SEQ ID NO: 1 coding for the ⁇ -chain and/or comprising the nucleic acid sequence of SEQ ID NO: 2 coding for the ⁇ -chain of said TCR.
  • a TCR of the present invention and a nucleic acid sequence encoding the same may comprise only one of the ⁇ -chain or ⁇ -chain sequences as defined herein (in combination with a further ⁇ -chain or ⁇ -chain, respectively) or may comprise both chains.
  • nucleic acid refers to a naturally- occurring nucleic acid that is not immediately contiguous with both of the sequences with which it is immediately contiguous (one on the 5'end and one on the 3'end) in the naturally- occurring genome of the cell from which it is derived.
  • a nucleic acid can be, without limitation, a recombinant DNA molecule of any length, provided one of the nucleic acid sequences normally found immediately flanking that recombinant DNA molecule in a naturally-occurring genome is removed or absent.
  • a nucleic acid includes, without limitation, a recombinant DNA that exists as a separate molecule (e.
  • an isolated nucleic acid can include a recombinant DNA molecule that is part of a hybrid or fusion nucleic acid sequence.
  • nucleic acid as used herein also includes artificially produced DNA or RNA sequences, such as those sequences generated by DNA synthesis based on in silico information.
  • the nucleic acids of the invention can comprise natural nucleotides, modified nucleotides, analogs of nucleotides, or mixtures of the foregoing as long as they are capable of causing the expression of a polypeptide in vitro, and preferably, in a T cell.
  • the nucleic acids of the invention are preferably RNA, and more preferably DNA.
  • the present invention also comprises derivatives of the above described nucleic acid molecules, wherein, related to the above SEQ ID NO: 1 and 2, the sequence has been altered by additions, deletions and/or substitutions and wherein the tumor antigen recognizing characteristics are maintained or improved. Irs other v*ordt>, ibc iumor antigen recognizing are at least maintained.
  • such a derivative is coding for the ⁇ - or ⁇ -chain, wherein the chain has been altered by one or more additions or deletions of from 1-15 amino acids, the additions or deletions being outside the CDR3 region of each chain, and/or by conservative substitutions of from 1-15 amino acids. It is noted in this connection that also the CDR3 region may be altered, but to a lesser extent. The definition of those amendments is indicated below for the derivatives of fragments coding for the CDR3 region.
  • Useful changes in the overall nucleic acid sequence in particular are related to codon optimization and the addition of epitope tags, which will be explained in detail below. Such codon optimization can include optimization of expression levels, optimization of avidity for target cells, or both.
  • the alterations should not diminish or alter the ability of the encoded polypeptide to form part of a TCR that recognizes tumor associated antigens in the context of an MHC, but should facilitate destruction of a cancer cell, and preferably facilitate the regression of a tumor, or other cancerous state.
  • alterations can be made which lead to conservative substitutions within the expressed amino acid sequence.
  • These variations can be made in complementarity determining and non-complementarity determining regions of the amino acid sequence of the TCR chain that do not affect function.
  • additions and deletions should not be performed in the CDR3 region (for example an addition of epitope tags).
  • codons encoding positively-charged residues H, K, and R are substituted with codons encoding positively-charged residues
  • codons encoding negatively- charged residues D and E are substituted with codons encoding negatively-charged residues
  • codons encoding neutral polar residues C, G, N, Q, S, T, and Y are substituted with codons encoding neutral polar residues
  • codons encoding neutral non-polar residues A, F, I, L, M, P, V, and W are substituted with codons encoding neutral non-polar residues.
  • the present invention provides fragments of the above nucleic acid molecules, coding for a CDR3 region of a TCR recognizing a tumor antigen and having the nucleic acid sequence of SEQ ID NO: 3 or 4 or coding for the amino acid sequences of SEQ ID NO : 5 or 6. Alterations in the CDR3 region will be performed according to the considerations described below.
  • the invention further provides derivatives wherein the CDR3 region has been altered by one or more additions and/or deletions of an overall number of from 1-5 amino acids, but not more than 1-3 contiguous amino acids and/or conservative substitutions of from 1-6 amino acids and wherein the tumor antigen recognizing characteristics are maintained or improved.
  • additions or deletions may be performed to an extent that 1-5 amino acids are added or deleted in the CDR3 region. If more then one addition or deletion is performed, the overall number of added or deleted amino acids may not exceed 5 amino acids. Further, one single addition or deletion at one site may only be in the range of 1-3 amino acids, i.e. 1-3 contiguous amino acids, since the ligand binding capacity might be deteriorated by performing larger additions/deletions.
  • a preferred derivative of the nucleic acid molecule encoding the ⁇ - or ⁇ -chain of said TCR is one, wherein the original sequence of SEQ ID NO: 1 and 2 has been altered by codon optimization.
  • a preferred example of such a derivative coding for the V(D)J regions of a TCR that recognizes a tumor antigen is the nucleic acid sequence of SEQ ID NO: 7 coding for the ⁇ -chain and the nucleic acid sequence of SEQ ID NO: 8 coding for the ⁇ -chain of said TCR.
  • Codon optimization is a generic technique to achieve optimal expression of a foreign gene in a cell system. Selection of optimum codons depends on codon usage of the host genome and the presence of several desirable and undesirable sequence motifs. It is noted that codon optimization will not lead to an altered amino acid sequence and, thus, will not fall under the definition of a conservative substitution as contained in this application.
  • the tumor antigen is tyrosinase.
  • Tyrosinase expressing malignancies still have a high incidence, for example, around 160,000 new cases of melanoma are diagnosed worldwide each year. According to a report issued by WHO, about 48,000 melanoma related deaths occur worldwide per year.
  • tyrosinase is a suitable tumor antigen which can serve as a target for tumor treatment.
  • the present invention is directed to a TCR, preferably a soluble TCR, encoded by a nucleic acid molecule as defined above or comprising the amino acid sequences of SEQ ID NO: 5 and/or 6.
  • Said TCR preferably is present in the form of a functional TCR ⁇ -and/or ⁇ -chain fusion protein, comprising: a) at least one epitope-tag, and b) the amino acid sequence of an ⁇ and/or ⁇ chain of a TCR as defined above or encoded by a nucleic acid molecule as outlined above, wherein said epitope-tag is selected from i) an epitope-tag added to the N- and/or C-terminus of said ⁇ - and/or ⁇ -chain, or added into the ⁇ - and/or ⁇ -chain sequence, but outside the CDR3 region, ii) an epitope-tag inserted into a constant region of said ⁇ - and/or ⁇ -chain, and iii) an epitope-tag replacing a number of amino acids in a constant region of said ⁇ -and/or ⁇ - chain.
  • a functional TCR ⁇ -and/or ⁇ -chain fusion protein comprising: a) at least one
  • Epitope tags are short stretches of amino acids to which a specific antibody can be raised, which in some embodiments allows one to specifically identify and track the tagged protein that has been added to a living organism or to cultured cells. Detection of the tagged molecule can be achieved using a number of different techniques. Examples of such techniques include: immunohistochemistry, immunoprecipitation, flow cytometry, immunofluorescence microscopy, ELISA, immunob lotting ("Western"), and affinity chromatography. Epitope tags add a known epitope (antibody binding site) on the subject protein, to provide binding of a known and often high-affinity antibody, and thereby allowing one to specifically identify and track the tagged protein that has been added to a living organism or to cultured cells.
  • a "functional" T-cell receptor (TCR) ⁇ - and/or ⁇ - chain fusion protein shall mean an ⁇ - and/or ⁇ -chain fusion protein that, although the chain includes the epitope-tag and/or has a tag attached to it, maintains at least substantial fusion protein biological activity in the fusion.
  • TCR T-cell receptor
  • ⁇ - and/or ⁇ -chain of a TCR this shall mean that both chains remain able to form a T-cell receptor (either with a non- modified ⁇ - and/or ⁇ -chain or with another inventive fusion protein ⁇ - and/or ⁇ -chain) which exerts its biological function, in particular binding to the specific peptide-MHC complex of said TCR, and/or functional signal transduction upon peptide activation.
  • TCR functional T-cell receptor
  • said epitope-tag has a length of between 6 to 15 amino acids, preferably 9 to 11 amino acids.
  • TCR T-cell receptor
  • TCR T-cell receptor
  • ⁇ - and/or ⁇ -chain fusion protein comprises two or more epitope-tags, either spaced apart or directly in tandem.
  • Embodiments of the fusion protein can contain 2, 3, 4, 5 or even more epitope-tags, as long as the fusion protein maintains its biological activity/activities ("functional").
  • TCR T-cell receptor
  • said epitope-tag is selected from, but not limited to, CD20 or Her2/neu tags, or other conventional tags such as a myc-tag, FLAG-tag, T7-tag, HA (hemagglutinin)-tag, His-tag, S-tag, GST-tag, or GFP -tag.
  • myc, T7, GST, GFP tags are epitopes derived from existing molecules.
  • FLAG is a synthetic epitope tag designed for high antigenicity (see, e.g., U.S. Pat. Nos. 4,703,004 and 4,851,341).
  • the myc tag can preferably be used because high quality reagents are available to be used for its detection.
  • Epitope tags can of course have one or more additional functions, beyond recognition by an antibody. The sequences of these tags are described in the literature and well known to the person of skill in art.
  • said fusion protein may be for example selected from two myc-tag sequences that are attached to the N-terminus of an ⁇ -TCR-chain and/or 10 amino acids of a protruding loop region in the ⁇ -chain constant domain being exchanged for the sequence of two myc-tags.
  • the inventors inserted an amino acid sequence that corresponds to a part of the myc protein (myc-tag) at several reasonable sites into the structure of a T cell receptor and transduced this modified receptor into T cells (see examples below).
  • myc-tag a part of the myc protein
  • TCR fusion proteins may be used in a method for selecting a host cell population expressing a fusion protein selected from the group consisting of a fusion protein comprising a) at least one epitope-providing amino acid sequence (epitope-tag), and b) the amino acid sequence of an ⁇ - and/or ⁇ -chain of a TCR as defined above, wherein said epitope-tag is selected from an epitope-tag added to the N- and/or C-terminus of said ⁇ - and/or ⁇ -chain or added into the ⁇ - and/or ⁇ -chain sequence, but outside the CDR3 region, an epitope-tag inserted into a constant region of said ⁇ - and/or ⁇ -chain, and an epitope-tag replacing a number of amino acids in a constant region of said ⁇ - and/or ⁇ -chain; and a TCR comprising at least one fusion protein as above on the surface of the host cell; comprising contacting host cells in a sample with a
  • the present invention further provides an immunoglobulin molecule, anticaline, TCR ⁇ / ⁇ chain having a CDR3 region as defined herein (or a derivative thereof) inserted.
  • the invention is directed to a T cell expressing a TCR as defined herein or a TCR comprising one of the CDR3 regions as defined above.
  • the invention provides a vector, preferably a plasmid, shuttle vector, phagemide, cosmid, expression vector, retroviral vector, adenoviral vector or particle and/or vector to be used in gene therapy, which comprises one or more of the nucleic acids as disclosed above.
  • a "vector” shall mean a nucleic acid molecule as introduced into a host cell, thereby producing a transformed host cell.
  • a vector may include nucleic acid sequences that permit it to replicate in a host cell, such as an origin of replication.
  • a vector may also include one or more selectable marker genes and other genetic elements known to those of ordinary skill in the art.
  • a vector preferably is an expression vector that includes a nucleic acid according to the present invention operably linked to sequences allowing for the expression of said nucleic acid.
  • a fourth aspect provides a cell, preferably a peripheral blood lymphocyte (PBL) which has been transformed with the above vector.
  • PBL peripheral blood lymphocyte
  • the step of cloning the T cell receptor (TCR) of the isolated T cells and/or expressing the TCR transgenes in PBMC can be done according to established methods such as those described in Engels et al, 2005.
  • the present invention provides a pharmaceutical composition which comprises a TCR, a T cell or cell (PBL) as defined above and a pharmaceutically acceptable carrier.
  • Those active components of the present invention are preferably used in such a pharmaceutical composition, in doses mixed with an acceptable carrier or carrier material, that the disease can be treated or at least alleviated.
  • a composition can (in addition to the active component and the carrier) include filling material, salts, buffer, stabilizers, solubilizers and other materials, which are known state of the art.
  • pharmaceutically acceptable defines a non-toxic material, which does not interfere with effectiveness of the biological activity of the active component.
  • the choice of the carrier is dependent on the application.
  • the pharmaceutical composition can contain additional components which enhance the activity of the active component or which supplement the treatment. Such additional components and/or factors can be part of the pharmaceutical composition to achieve synergistic effects or to minimize adverse or unwanted effects.
  • An appropriate application is a parenteral application, for example intramuscular, subcutaneous, intramedular injections as well as intrathecal, direct intraventricular, intravenous, intranodal, intraperitoneal or intratumoral injections.
  • the intravenous injection is the preferred treatment of a patient.
  • the pharmaceutical composition is an infusion or an injection.
  • An injectable composition is a pharmaceutically acceptable fluid composition comprising at least one active ingredient, e.g., an expanded T-cell population (for example autologous or allogenic to the patient to be treated) expressing a TCR.
  • the active ingredient is usually dissolved or suspended in a physiologically acceptable carrier, and the composition can additionally comprise minor amounts of one or more non-toxic auxiliary substances, such as emulsifying agents, preservatives, and pH buffering agents and the like.
  • Such injectable compositions that are useful for use with the fusion proteins of this disclosure are conventional; appropriate formulations are well known to those of ordinary skill in the art.
  • the present invention is directed to a method of treating a patient in need of adoptive cell therapy, said method comprising administering to said patient a pharmaceutical composition as defined above to said patient.
  • the patient to be treated preferably belongs to the group of HLA- A2 positive patients.
  • said patient suffers from a disease involving malignant cells expressing tyrosinase, preferably melanoma, glioma, glioblastoma, and/or rare tumors of ectodermal origin.
  • tyrosinase preferably melanoma, glioma, glioblastoma, and/or rare tumors of ectodermal origin.
  • Fig. 1 Screening of clones obtained from limiting dilution cultures after DC priming. T cells were primed with dendritic cells expressing HLA- A2 and tyrosinase RNA. After two rounds of priming in vitro, cells were cloned by limiting dilution. 14 to 28 days later T cell clones showing adequate growth in individual culture wells were identified by light microscopy. Aliquots of growing clones were obtained and tested in a standard 51 Cr release assay to measure their killing activity against two melanoma target cell lines. Mel-A375 cells express HLA-A2 but not tyrosinase.
  • Mel-93.04A12 cells express HLA-A2 and tyrosinase, so they can form the ligands recognized by HLA-A2 -restricted, tyrosinase peptide-specific T cells. If Mel-A375 cells are recognized by T cell clones, this means the clones are alloreactive and recognize HLA- A2 independent of tyrosinase peptide (ie clone T41 and T42). If the T cell clones only recognize Mel-93.04A12, then they should have specificity for HLA-A2 -tyrosinase peptide ligands (i.e. T58, T43).
  • Percentage specific lysis mediated by various T cell clones (listed on x-axis) is given for the two target melanoma cell lines.
  • the arrow designates clone T58 which shows strong killing of Mel-93.04A12 but not of Mel-A375. This clone was selected for further characterization based on its strong growth capacity.
  • Fig. 2 Comparison of clones T58 and IVS-B
  • Fig. 2a Cytotoxic activity directed against melanoma cell lines.
  • the killing capacity of clone T58 was compared with that of clone IVS-B, derived from a melanoma patient, using as target cells the T2 cell line pulsed with synthetic tyrosinase- peptide for the amino acid sequence YMDGTMSQV in different molar concentrations, listed on the x-axis.
  • the % relative lysis is given on the y-axis.
  • the concentration of peptide that corresponds to 50% relative lysis is indicated by the crossing lines and shows that clone T58 can recognize substantially lower concentrations of peptide in comparison to clone IVS-B.
  • Fig. 2b Measurement of multimer binding and off-rates.
  • the two clones were incubated with multimers to determine the percentage of positive cells at time 0 h. Both clones bound multimer on 100% of the cells. Multimer was washed out and the clones were incubated in medium containing HLA-A2-specific antibody. When multimers are released from the cell surface, they are captured by the antibody and can not rebind to the cells. The percent multimer-positive cells were reanalyzed at 1 h and 2 h.
  • Fig. 2c Interferon-gamma secretion after stimulation with melanoma cell lines.
  • Fig. 2d Cytotoxic activity against melanoma cell lines.
  • Fig. 3 Recognition of primary melanoma tumor cells by clone T58 and IVS-B.
  • control curves represent untransfected primary tumor cells stained with HLA-A2-specific monoclonal antibody (left histogram) or melanoma cell lines stained with isotype control antibody.
  • HLA- A2 protein expression on RNA-transfected primary tumor cells was detected 1O h after electroporation.
  • IVS-B The capacity of the patient-derived T cell clone (IVS-B), and T cell clone T58 to secrete IFN- ⁇ or
  • release perform in co- culture with the melanoma cells shown above was measured in ELISPOT assays.
  • Fig. 4 Transfer of antigen specificity by TCR retroviral gene transfer
  • the human TCR- deficient T cell line Jurkat76 9 was transduced with the TCR of the T cell clone T58.
  • TCR- expression was detected using tyrosinase-peptide-specific HLA-multimers.
  • TCR expression was only detected in Jurkat76 cells tranduced with TCR-T58 (right histogram) and not in untransduced Jurkat76 cells (left histogram)
  • PBL of a healthy donor were retrovirally transduced with TCR-T58. After 10 days, untransduced and TCR-transduced PBL were analysed for tyrosinase TCR-expression using specific HLA-multimers.
  • Multimer staining is shown on the x-axis and CD8 staining on the y-axis.
  • the percentage of multimer + CD8 + T cells is displayed in the upper right quadrant,
  • (c) Functionality of TCR-transduced PBL was measured using a standard IFN- ⁇ release assay.
  • T2 cells loaded with graded amounts of tyrosinase369-377 peptide (10 ⁇ 12 M - 10 "5 M) were used as target cells at a fixed effector to target cell ratio of 1 : 1.
  • Untransduced PBL served as a control and showed no tyrosinase- peptide specific IFN- ⁇ release (data not shown).
  • Fig. 5 Transfer of specificity of T58 and IVS-B for HLA- A2 and tyrosinase-peptide YMDGTMSQV by TCR retroviral gene transfer, (a) PBL of a healthy donor were retrovirally transduced with the patient-derived TCR-IVS-B or the TCR-T58. After 11 days, untransduced and TCR-transduced PBL were analysed for tyrosinase TCR-expression using specific HLA-multimers. Multimer staining is shown on the x-axis and CD8 staining on the y-axis.
  • T2 cells loaded with graded amounts of tyrosinase369-377 peptide (10 "11 M - IO "5 M) or with 10 "5 M irrelevant influenza matrix proteinss- ⁇ were used as target cells at a fixed effector to target cell ratio of 1 :1.
  • Untransduced PBL served as a control and showed no tyrosinase-peptide specific IFN- ⁇ release (data not shown).
  • Fig. 6 Tyrosinase peptide-specific CTL recognition of tumor cell lines and primary melanoma tumor cells.
  • Columns represent the amount of IFN- ⁇ (pg/ml) secreted by self- restricted Dl 15 CTL and allo-restricted T58 CTL in co-culture with a panel of tumor cell lines from left to right: MaCaI (HLA-A2-tyrosinase-); SK-Mel-28 (HLA-A2-tyrosinase+); Mel-A375, RCC-26, PancTu 1, MaCal/A2, and UTS CC 1588 (all HLA-A2+tyrosinase-); Mel-624.38, Mel-93.04A12, SK-Mel-23, SK-Mel-29 and WM-266-4 (all HLA- A2+tyrosinase+).
  • T cells designates CTL without stimulating cells.
  • the HLA- A2+tyrosinase- tumor cell lines Mel-A375, RCC-26 and MaCal/A2 were exogenously loaded with either 10-5 M irrelevant flu peptide or 10-5 M tyrosinase peptide YMD and IFN- ⁇ secretion was measured by ELISA and given as pg/ml.
  • Fig. 7 Transfer of antigen specificity by retroviral transfer of TCR-Dl 15 and TCR-T58.
  • PBL of a healthy donor were transduced with TCR-Dl 15 or TCR-T58.
  • Specificity of recognition was assessed by IFN- ⁇ release following co-culture with the tumor cell lines from left to right: MaCaI (HLA-A2-tyrosinase-); SK-Mel-28 (HLA-A2-tyrosinase+); MeI- A375, RCC-26, PancTu 1, MaCal/A2, and UTS CC 1588 (all HLA-A2+tyrosinase-); Mel- 624.38, Mel-93.04A12, SK-Mel-23, SK-Mel-29 and WM-266-4 (all HLA-A2+tyrosinase+).
  • T designates CTL without stimulating cells.
  • the HLA-A2+tyrosinase- tumor cell lines MeI- A375, RCC-26 and MaCal/A2 were exogenously loaded with either 10-5 M irrelevant flu peptide or 10-5 M tyrosinase peptide YMD and IFN- ⁇ secretion was measured by ELISA and given as pg/ml.
  • Fig. 8 Transfer of antigen specificity by retroviral transfer of TCR-Dl 15 and TCR-T58.
  • A PBL of a healthy donor were transduced with TCR-Dl 15 or TCR-T58.
  • Unsorted TCR- transduced PBL were analyzed on day 10 for transgenic TCR-expression using irrelevant B7-pp65 and A2-pp65 multimers and specific A2-tyr multimers. Untransduced PBL showed no multimer binding (0.1%, data not shown). Percentages of multimer+CD8+ T cells are displayed in the upper right quadrant.
  • B) and (C) show the IFN- ⁇ release of unsorted TCR- transduced PBL following stimulation with T2 cells loaded with graded amounts of tyrosinase peptide (10-12 M - 10-5 M) at a ratio of 2:1. In (B) the relative IFN- ⁇ release is displayed in percent and in (C) the specific IFN- ⁇ release is presented as pg/ml.
  • Fig. 9 TCR transfer retains differences in cytokine profile.
  • A-D On the left hand side the cytokine release of TCR-transduced PBL in co-culture with the melanoma lines Mel-A375 (HLA-A2+tyrosinase-) and Mel-624.38 (HLA-A2+tyrosinase+) is depicted, on the right hand side the corresponding cytokine release after stimulation with T2 cells loaded with graded amounts of tyrosinase peptide (10-12 M - 10-5 M) is shown. Untransduced PBL (A) showed no peptide-specific cytokine release.
  • cytokine secretion was measured: IFN- ⁇ (A), IL-2 (B), TNF- ⁇ (C) and MIP- l ⁇ (D).
  • IFN- ⁇ A
  • IL-2 B
  • TNF- ⁇ C
  • MIP- l ⁇ D
  • the levels of cytokine secretion for all four cytokines were higher when PBL transduced with the allo-restricted TCR-T58 were used. Since untransduced PBL secreted very high levels of MIP-I ⁇ in response to T2 cells the peptide titration for this cytokine could not be evaluated.
  • the inventors prepared stimulating dendritic cells (DC) from an HLA- A2 -negative healthy donor that co-expressed allogeneic HLA-A*0201 -molecules and tyrosinase protein using mature DC that were electroporated with in vitro transcribed (ivt)-RNA for tyrosinase and HLA-A2, as described 1 ' 2 . These DC were used to prime purified, autologous CD8 + T cells using two rounds of stimulation with freshly prepared DC.
  • DC dendritic cells
  • CD8 + T cells with T cell receptors (TCR) recognizing HLA-A2-tyrosinase369-377- peptide complexes were stained using a tyrosinase 369 - 377 /HLA-A*0201-multimer 3 .
  • CD8 multimer + cells were isolated by fluorescence activated cell sorting. Sorted cells were cloned in limiting dilution cultures and isolated clones showing HLA-A2/tyrosinase-peptide specificity were expanded using antigen- independent stimulation 4 .
  • the T cell clone T58 was identified in an initial screen as having good functional activity (Figure 1).
  • T58 was isolated from an HLA-A*0201 -negative donor it represents an allo- restricted T cell clone that did not undergo negative selection in vivo.
  • the activity of the T58 clone was compared with the IVS-B clone that was isolated from a patient with metastatic melanoma 5 .
  • This clone recognizes exactly the same HLA-A2/tyrosinase peptide ligand as clone T58 but it is self-restricted since it was activated in vivo in the patient who was HLA-A*0201 -positive.
  • This patient-derived T cell clone represents an example of T cells that are available in the peripheral repertoire that have undergone negative selection against self-peptides/self-MHC-molecules in the thymus in vivo.
  • the allo-restricted T cell clone T58 required substantially less peptide to be activated by peptide-pulsed T2 cells than clone IVS-B (6.OxIO "10 M vs. 3.OxIO "8 M) (Fig. 2a).
  • loss of multimer binding was measured over time (i.e. HLA-multimer off-rate).
  • a slower off-rate indicates that TCR- ligand interactions are more stable and of higher structural affinity 7 .
  • T cells were incubated for 1 h and 2 h without multimers in the presence of HLA-A2-specific antibody to prevent cellular re-association of released multimers.
  • the melanoma patient-derived T cell clone IVS-B showed an intermediate multimer binding: all cells were multimer + at 0 h and about 40% retained multimers at 1 and 2 h (Fig. 2b).
  • clone T58 had a slower off-rate, showing 74% positive binding at 1 h versus 41% for clone IVS-B and even at 2 h still had somewhat more multimer + cells (55% vs. 40%).
  • T cell clones were analyzed in an IFN- ⁇ release assay for function and specificity (Fig. 2c).
  • the clones were co-cultured with two melanoma cell lines that express HLA-A2 molecules but differ with respect to expression of tyrosinase protein: Mel-93.04A12 co- expresses both proteins (HLA- A2 + tyrosinase ) but Mel-A375 fails to express tyrosinase protein (HLA- A2 + tyrosinase " ) and therefore can not generate the MHC-peptide ligand seen by the T cell clones.
  • AlIo -restricted T cell clone T58 was induced to secrete a high level of IFN- ⁇ by the tyrosinase-expressing melanoma cell line, whereas only marginal cytokine secretion was seen with IVS-B cells (1,234 pg/ml vs. 106 pg/ml), demonstrating the vastly superior function of clone T58 in recognizing tumor cells expressing their HLA-A2- tyrosinase ligand. As expected, the clones showed no detectable IFN- ⁇ secretion after stimulation with Mel- A375 cells, demonstrating the specificity for HLA- A2 and tyrosinase expression for tumor cell recognition.
  • TCR alpha and beta chains of clone T58 were co-infected with the ⁇ -chain and ⁇ -chain retroviruses and transgenic TCR-expression was measured by multimer staining. TCR-T58 was expressed at a good level, demonstrating adequate quality of the separate retroviral supernatants (Fig. Aa).
  • PBL peripheral blood lymphocytes
  • Fig. Ab activated peripheral blood lymphocytes
  • TCR-T58 transduced PBL could also respond specifically to stimulation by melanoma cell lines that expressed HLA-A2 and tyrosinase (Fig. Ad). They did not respond to tumor cells that did not express HLA- A2 or tyrosinase, again demonstrating the specificity of HLA- A2 -tyrosinase ligands for T58 recognition.
  • Bi-cistronic retroviral vectors were also prepared encoding the ⁇ -chain and ⁇ -chains of the TCR of IVS-B cells and used to transduce activated PBL.
  • the same activated PBL were transduced with bi-cistronic retroviral vectors encoding the two chains of TCR- T58.
  • PBL expressing the corresponding receptors were identified by co-staining for CD 8 and multimer and showed low numbers of positive cells. (Fig. 5a)
  • PBL transduced with TCR-T58 released high amounts of IFN- ⁇ following stimulation with T2 cells pulsed with graded amounts of tyrosinase-peptide.
  • Table 1 shows the genetic information regarding the use of VJ and VDJ gene segments by the alpha and beta chains of TCR-T58, respectively.
  • the CDR3 regions, according to IMGT, are presented as nucleotide sequences and amino acid sequences. Also shown are the codon optimized sequences for the full VJ and VDJ regions.
  • the human melanoma cell lines Mel-A375 (HLA- A2 + , tyrosinase " ; CRL-1619, American Type Culture Collection (ATCC), Bethesda, MD), Mel-93.04A12 (HLA-A2 + , tyrosinase + , gift of P. Schrier, Department of Immunohematology, Leiden University Hospital, The Netherlands), Mel-624.38 10 (HLA-A2 + , tyrosinase + , gift of M. C.
  • SK-Mel-28 HLA- A2 " , tyrosinase + ; MTB-72, ATCC
  • T2 lymphoid cell line
  • human TCR-deficient Jurkat76 9 T cell line were cultured in RPMI 1640 medium supplemented with 12% fetal bovine serum (FBS), 2 mM L-glutamine and 1 mM sodium-pyruvate and non-essential amino acids.
  • the HLA- A*0201 -restricted tyrosinase369-377 peptide-specific melanoma patient-derived IVS-B T cell clone was cultured as described 5 .
  • plasmid pCDM8-HLA-A2 with HLA-A*0201 cDNA and the pZeoSV2+/huTyr with tyrosinase cDNA were linearized and used as in vitro transcription templates to produce RNA with the aid of the mMESSAGE mMACHINE T7 kit (Ambion, Austin, TX) according to the manufacturer's instructions.
  • PBL Peripheral blood lymphocytes
  • VLE very low endotoxin
  • DC medium human serum
  • Mature DC were prepared from adherent monocytes and transfected with ivt-RNA via electroporation as previously described 2 .
  • DC of HLA-A2 " donors were co-transfected with 24 ⁇ g tyrosinase ivt-RNA and 48 ⁇ g HLA- A2 ivt-RNA.
  • autologous CD8 + T lymphocytes were enriched from PBL via negative selection using a commercial kit according to the manufacturer's instructions (CD8 + T cell Isolation Kit II (human), Miltenyi, Bergisch Gladbach, Germany).
  • Co-cultures were initiated 1 O h after DC electroporation in 24-well plates (TPP, Trasadingen, Switzerland) by adding IxIO 5 RNA- pulsed DC to IxIO 6 CD8 + T cells in RPMI 1640, supplemented with 10% heat-inactivated human serum, 4 mM L-glutamine, 12.5 mM HEPES, 50 ⁇ M ⁇ -mercaptoethanol and 100 U/ml penicillin/streptomycin (T cell medium).
  • IL-7 (5 ng/ml) (Promokine, Heidelberg, Germany) was added on day 0 and 50 U/ml IL-2 (Chiron Behring, Marburg, Germany) was added after 2 days and then on every 3 rd subsequent day. Addition of IL-2 was delayed to decrease proliferation of non-specific CD8 + T cells 4 .
  • the 2 nd stimulation of primed T cells was made after seven days using freshly prepared RNA-pulsed DC.
  • HLA- A2 -restricted tyrosinase-specific T cells were detected by staining with a PE-labeled HLA- A*0201/htyr 36 9-377 peptide/human ⁇ 2 m multimer 11 , anti-CD8-APC antibody (clone RPA-T8, BD Pharmingen, Franklin Lakes, NJ) and propidium iodide (PI: 2 ⁇ g/ml).
  • PI propidium iodide
  • CD8- APC antibody was then added at 1/50 for an additional 25 min. After staining cells were washed twice and diluted in PBS + 0.5% human serum with PI for sorting. 20-5OxIO 6 total cells per priming culture were stained for sorting. Pi-negative cells were gated and CD8 multimer + T cells were sorted on a FACSAria cell sorter (BD Biosciences) with a 70 ⁇ m nozzle, at a rate of 15,000 events/s.
  • BD Biosciences FACSAria cell sorter
  • Multimer-sorted T cells were cloned by limiting dilution. Clones were plated in 96-well round-bottom plates (TPP) in 200 ⁇ l/well T cell medium. 50 IU/ml IL-2 was supplemented every 3 days with 5 ng/ml IL-7 and 10 ng/ml IL- 15 (PeproTech Inc., Rocky Hill, NJ) every 7 days.
  • T cell clones were stimulated non-specifically with anti-CD3 antibody (0.1 ⁇ g/ml; OKT-3) and provided with 1x10 5 feeder cells per 96-well, consisting of irradiated (50 Gy) PBL derived from a pool of five unrelated donors and IxIO 4 irradiated (150 Gy) EBV- transformed allogeneic B-LCL every two weeks.
  • Proliferating T cells were transferred into 24-well plates (TPP) and cultured in 1.5 ml T cell medium plus cytokines.
  • IxIO 6 allogeneic irradiated PBL and 1x10 5 irradiated EBV-transformed allogeneic B-LCL were added per well as feeder cells in 24-well plates. Clonality was determined by TCR-beta-chain receptor analysis, as described 12 .
  • T2 cells were incubated at 37°C and 5% CO 2 for 2 h with 10 ⁇ g/ml human ⁇ 2 -microglobulin (Calbiochem, San Diego, CA) and titrating amounts, ranging from 10 "5 M to 10 "12 M, of the tyrosinase peptide YMD (tyrosinase369-377 YMDGTMSQV, Metabion, Martinsried, Germany).
  • T2 cells pulsed with 10 "5 M influenza peptide GIL (influenza matrix protein58_66 GILGFVTL, Metabion) served as negative control. After washing, peptide-loaded T2 cells were used as target cells in cytotoxicity or IFN- ⁇ -release assays.
  • T cell clones (2x10 cells in 100 ⁇ l) were incubated with the respective melanoma cell lines or peptide-pulsed T2 cells (IxIO 4 cells in 100 ⁇ l). Culture supernatants were harvested after 24 h co-culture and assessed by a standard ELISA using the OptEIATM Human IFN- ⁇ Set (BD Biosciences Pharmingen). Cytotoxicity assay
  • Cytotoxic activity of T cell clones was analysed in a standard 4 h 51 -chromium release assay. Melanoma cells or peptide-loaded T2 cells were used as target cells. Briefly, 1x10 6 target cells were labeled with 100 ⁇ Ci Na 2 51 CrO 4 (ICN Biochemicals, Irvine, CA) for 1-1.5 h. 51 Cr-labeled target cells were cultured with T cells in 100 ⁇ l/well RPMI 1640 with 12% FC S in V-bottom 96-well tissue culture plates (Greiner, Solingen, Germany). For determination of functional avidity IxIO 4 T cells were added to IxIO 3 peptide-pulsed T2 cells loaded with titrated amounts of peptide, giving a constant E:T of 10:1.
  • Antibody pre-coated PVDF plates (Mabtech AB, Nacka, Sweden) were incubated at 37°C in CTL TestTM medium (Cellular Technology Ltd., Cleveland, Ohio) for 2 h to block unspecific binding.
  • CTL TestTM medium Cellular Technology Ltd., Cleveland, Ohio
  • IFN- ⁇ ELISPOT plates were pre-coated with the IFN- ⁇ -specific capture antibody clone 1-DlK; for perform ELISPOT plates were pre-coated with the perforin-specific capture antibody (clone Pf-80/164; Mabtech AB).
  • Primed T cells were washed with CTL WashTM Supplement culture medium (Cellular Technology Ltd) and IxIO 3 responder T cells were stimulated with 5xlO 3 melanoma cells in 150 ⁇ l CTL TestTM medium and 24 h later assessed in IFN- ⁇ ELISPOT or 48 h later in perform ELISPOT.
  • TCR identification of tumor-specific T cell clones part of the TCR ⁇ - and TCR ⁇ -chain sequences including the complementary determining region (CDR3) was amplified by PCR using a panel of TCRV ⁇ and TCRV ⁇ primers combined with the respective constant region primer as described 13 .
  • the TCR ⁇ and TCR ⁇ chain genes of T cell clones T58 and IVS-B were amplified by PCR with gene specific primers and cloned into the retroviral vector MP71PRE 8 via Notl and EcoRI restriction sites.
  • Both chains of human TCR-T58 (V ⁇ 7, V ⁇ 23) and TCR-IVS-B (V ⁇ 3, V ⁇ l4) were constructed as single-TCR gene vectors or double-TCR gene vectors (pMP71-T58 ⁇ and pMP71-T58 ⁇ , pMP71-IVS-B ⁇ and pMP71- IVS-B ⁇ ; pMP71-T58 ⁇ -P2A-T58 ⁇ and pMP71-IVS-B ⁇ -P2A-IVS-B ⁇ ).
  • Retroviral vector plasmids were co-transfected into 293T cells with expression plasmids encoding Moloney MLV gag/pol and MLV-IOAl env gene to produce amp ho tropic ML V-pseudo typed retroviruses as described 14 .
  • the human TCR-deficient T cell line Jurkat76 and PBL were transduced as reported 14 .
  • Jurkat76 cells (5 days after transduction) and PBL (10 days after transduction) were stained using PE-labeled HLA-A* 020 l/htyr 36 9-377 peptide/human ⁇ 2 m multimer and anti-CD8-FITC antibody.
  • Control values for peptide-stimulated untransduced PBL were subtracted from values of transduced cells at each peptide concentration and then adjusted to comparable numbers of total TCR-transgenic cells.
  • T-cell receptor analysis of the tyrosinase-specif ⁇ c clone T58 part of the TCR alpha- chain and beta-chain containing the CDR3 region was amplified by RT-PCR using a panel of TCR Va and TCR V ⁇ primers combined with a respective TCR constant region primer. Products were sequenced and assigned according to IMGT (Table 1 ; IMGT, the international ImMunoGeneTics information system ® , http://imgt.cines.fr).
  • Antibody-tags for example myc-tags 15 (Patent Application number: 06014606.5-1212) or other modifications, for example a CD20 epitope, can be introduced in any position, i.e. the N-terminus of the TCR ⁇ -chain, that is recognized by the depleting antibody and does not interfere with TCR- functionality.
  • TCR alpha-chain (VJ region), TCR beta-chain (VDJ region) and CDR3 lenghts are designated according to IMGT (IMGT, the international ImMunoGeneTics information system ® , http ://imgt.cines. fr)
  • Example 1 data are provided that compared two T cell clones that specifically recognize a peptide derived from tyrosinase (ie YMDGTMSQV hereafter referred to as YMD) presented by HLA-A*0201 molecules.
  • the T cell clone T58 was an allo-restricted, peptide- specif ⁇ c T cell clone derived from an HLA-A2-negative donor.
  • the T cell clone IVS-B was derived from an HLA-A*0201 -positive patient who suffered from metastatic melanoma. This melanoma expressed tyrosinase.
  • comparisons have been extended to include an example of a T cell clone, Dl 15, which is also derived from an HLA-A* 0201 -positive individual and recognizes the same YMD peptide.
  • Dl 15 was generated in vitro using responding T cells derived from the blood of a healthy individual. Therefore, there have been no potential negative impacts on this T cell clone from a tumor environment (ie melanoma) in vivo.
  • Figure 6 shows a comparison of the pattern of the target cell recognition of the new clone Dl 15 and clone T58 which is the subject of this patent.
  • both Dl 15 and T58 show the same pattern of recognition, detected by secretion of interferon-gamma (y-axis), after co-cultivation with various tumor cell lines (x-axis and figure legend).
  • y-axis interferon-gamma
  • x-axis and figure legend Neither clone recognizes tumor cells that are HLA- A2 -negative but express tyrosinase, nor do they recognize tumor cells that are HLA- A2 -positive and tyrosinase negative.
  • both T cell clones recognize several tumor cell lines that are both HLA- A2 -positive and tyrosinase-positive.
  • the role of the YMD peptide in this recognition is shown by the finding that HLA-A2 -positive tumor cells that do not express tyrosinase from which the YMD peptide could be processed internally and transported to the cell surface by HLA-A2 molecules for presentation, can be loaded with synthetic YMD peptide, leading to their recognition by Dl 15 and T58.
  • both clones show the same specificity for the YMD peptide presented by HLA- A2 molecules.
  • the TCR of clone Dl 15 and T58 were expressed as recombinant proteins in activated recipient lymphocytes (Figure 7). When these TCR-transduced lymphocytes were retested with the same panel of target cells, they showed the same specificity pattern as the original T cell clones, demonstrating that the TCR recognition was responsible for the results seen in Figure 6. Again, in Figure 7 it is demonstrated that the TCR of clone T58 shows superior recognition of the melanoma tumor cell lines that express HLA- A2 and tyrosinase and the YMD peptide-pulsed HLA-A2 -positive tumor cells.
  • FIG. 8A shows that the TCR-transduced lymphocytes show comparable levels of expression of the respective recombinant TCRs, with each transduced population having around 1 1 % of T cells that bind a MHC multimer comprised of HLA- A2 molecules presenting the YMD peptide. Such binding is not observed with control multimers that present other peptides derived from the pp65 protein of human cytomegalovirus.
  • T2 cells HLA- A2 -positive antigen-presenting cells
  • YMD peptide shown on the x-axis
  • Figure 8D shows another peptide-sensitivity assay, this time using peripheral blood mononuclear cells that have been pulsed with titrating amounts of YMD peptide (x-axis). Once again, the amounts of interferon-gamma released by lymphocytes expressing TCR- T58 are much greater compared with TCR-Dl 15. The arrows show that the first detection of cytokine secretion occurs with 1000-fold less peptide for TCR-T58 compared with TCR- D115.
  • Figures 8E and 8F demonstrate the specificity of the transduced lymphocyte populations for peptide-pulsed tumor cells (Figure 8E) or tumor cell lines expressing HLA-A2 and tyrosinase (Figure 8F). In all cases, recognition is superior by lymphocytes expressing TCR- T58 compared to TCR-Dl 15.
  • the superior secretion of cytokine is not limited to interferon-gamma.
  • the levels of secretion of interleukin-2, TNF-alpha and MIP-lbeta are also superior for TCR-T58. This is seen after stimulation of the TCR-transduced lymphocytes by tumor cells or by peptide- pulsed T2 cells (Figure 9).
  • the human melanoma cell lines Mel-A375 (HLA- A2 + , tyrosinase " ; CRL-1619, American Type Culture Collection (ATCC)), Mel-93.04A12 (HLA-A2 + , tyrosinase + ; gift of P. Schrier, Department of Immunohematology, Leiden University Hospital, The Netherlands), MeI- 624.38 1 and SK-Mel-23 (HLA-A2 + , tyrosinase + ; gift of M. C.
  • HLA-A*0201 transfectant of MaCaI (MaCal/A2) (HLA- A2 + , tyrosinase " , gift of E. Noessner, Institute of Molecular Immunology, Helmholtz Zentrum M ⁇ nchen, Germany), RCC-26 2 (HLA- A2 + , tyrosinase " ), PancTul (HLA- A2 + , tyrosinase " , gift of P. Nelson, Department for Biological Chemistry University Hospital LMU Kunststoff, Germany), UTS CC 1588 (HLA-A2 + , tyrosinase " , gift of M.
  • lymphoid cell line T2 (CRL- 1992, ATCC) were cultured in RPMI 1640 medium supplemented with 12% fetal bovine serum (FBS), 2 mM L-glutamine and 1 mM sodium-pyruvate and nonessential amino acids.
  • IxIO 6 T2 cells were incubated at 37°C and 5% CO 2 for 2 h with 10 ⁇ g/ml human ⁇ 2 -microglobulin (Calbiochem) and titrating amounts, ranging from 10 "5 M to 10 "11 M, of the tyrosinase peptide YMD (tyrosinase 36 9-377 YMDGTMSQV, Metabion).
  • T2 cells pulsed with 10 "5 M influenza peptide GIL (flu: influenza matrix protein58_66 GILGFVFTL, Metabion) served as the negative control.
  • PBMC peripheral blood mononuclear cells
  • Tumor cells were loaded with either 10 "5 M flu peptide or 10 "5 M tyrosinase peptide YMD as described for T2 cells. After washing, peptide-loaded T2 cells, PBMC or tumor cells were used as stimulating cells in IFN- ⁇ release assays.
  • CTL 2x10 cells in 100 ⁇ l
  • various tumor cell lines (1x10 4 cells in 100 ⁇ l)
  • peptide pulsing as described above.
  • Culture supernatants were harvested after 24 h co-culture and assessed by a standard ELISA using the OptEIATM Human IFN- ⁇ Set (BD Biosciences).
  • Data represent mean values with corresponding mean deviations calculated from duplicate determinations.
  • % relative IFN- ⁇ release the maximum IFN- ⁇ release was set to the reference value of 100% and corresponding values were calculated corresponding to this reference.
  • cytokine secretion in supernatants of co-culture of CTL with tumor cells and with or without tyrosinase peptide pulsed T2 cells (10 ⁇ 5 M) was measured using the multiplex protein array system technology (Bio-Rad Laboratories, Hercules, CA).
  • regions of the TCR ⁇ - and TCR ⁇ -chains encoding CDR3 were amplified by PCR using a panel of TCRV ⁇ and TCRV ⁇ primers in combination with respective constant region primers as described.
  • the full TCR ⁇ - and TCR ⁇ -chain genes of CTL clones T58 and Dl 15 were amplified by PCR using cDNA as template. Primer sequences will be provided on request.
  • the constant regions of both TCR chains were exchanged by the murine counterparts to increase the stability of the TCR.
  • TCR chains were linked by a 2A peptide linker (TCR ⁇ -P2 A-TCRa) 5 , codon-optimized (Geneart) 6 and cloned into the retroviral vector MP71PRE via Notl and EcoKI restriction sites.
  • Retroviral vector plasmids were co-transfected into 293T cells with expression plasmids encoding Moloney MLV gag/pol and MLV-10A1 env gene, respectively, to produce amphotropic MLV-pseudotyped retroviruses as described. 5
  • PBL were stained using PE-labeled A2-tyr multimer and FITC-labeled CD8-specific antibody.
  • Multimers presenting peptides derived from cytomegalovirus pp65 were used as controls: PE-labeled HLA-B7 pp654i7-427 (B7-pp65) multimers served as the HLA control and HLA- A2 pp65495_503 multimers as a peptide-specificity control.
  • an IFN- ⁇ release assay was performed using T2 cells or autologous PBMC loaded with graded amounts of tyrosinase peptide (10 ⁇ 12 M - 10 ⁇ 5 M) and the tumor cell lines MaCaI, SK-Mel-28, Mel-A375, RCC-26, PancTu 1, MaCal/A2, UTS CC 1588, Mel-624.38, MeI- 93.04A12, SK-Mel-23, SK-Mel-29 and WM-266-4 as stimulating cells at an E:T of 2:1.

Landscapes

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

Abstract

La présente invention concerne un récepteur de lymphocytes T (TCR) de forte affinité vis-à-vis d'un antigène associé à une tumeur, une molécule d'acide nucléique isolée codant ledit récepteur, un lymphocyte T exprimant ledit TCR et une composition pharmaceutique pouvant être employée dans le traitement de pathologies impliquant des cellules tumorales exprimant ledit antigène associé à une tumeur.
PCT/EP2009/065705 2008-11-24 2009-11-24 Récepteur de lymphocytes t de forte affinité et ses applications WO2010058023A1 (fr)

Priority Applications (11)

Application Number Priority Date Filing Date Title
CN200980154272.4A CN102272153B (zh) 2008-11-24 2009-11-24 高亲和力t细胞受体及其应用
ES09760145T ES2394180T3 (es) 2008-11-24 2009-11-24 Receptor de células T de alta afinidad y uso del mismo
AU2009317161A AU2009317161B2 (en) 2008-11-24 2009-11-24 High affinity T cell receptor and use thereof
US13/130,665 US8697854B2 (en) 2008-11-24 2009-11-24 High affinity T cell receptor and use thereof
EP09760145A EP2352756B1 (fr) 2008-11-24 2009-11-24 Récepteur de lymphocytes t de forte affinité et ses applications
CA2743669A CA2743669C (fr) 2008-11-24 2009-11-24 Recepteur de lymphocytes t de forte affinite et ses applications
JP2011536895A JP6069755B2 (ja) 2008-11-24 2009-11-24 高親和性ｔ細胞受容体およびその用途
DK09760145.4T DK2352756T3 (da) 2008-11-24 2009-11-24 Højaffin T-cellereceptor og anvendelse af denne
US14/224,525 US9862755B2 (en) 2008-11-24 2014-03-25 High affinity T cell receptor and use thereof
US15/822,970 US10626159B2 (en) 2008-11-24 2017-11-27 High affinity T cell receptor and use thereof
US16/820,856 US20200339653A1 (en) 2008-11-24 2020-03-17 High affinity t cell receptor and use thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP08020396 2008-11-24
EP08020396.1 2008-11-24

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US13/130,665 A-371-Of-International US8697854B2 (en) 2008-11-24 2009-11-24 High affinity T cell receptor and use thereof
US14/224,525 Division US9862755B2 (en) 2008-11-24 2014-03-25 High affinity T cell receptor and use thereof

Publications (1)

Publication Number Publication Date
WO2010058023A1 true WO2010058023A1 (fr) 2010-05-27

Family

ID=40521958

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2009/065705 WO2010058023A1 (fr) 2008-11-24 2009-11-24 Récepteur de lymphocytes t de forte affinité et ses applications

Country Status (9)

Country Link
US (4) US8697854B2 (fr)
EP (1) EP2352756B1 (fr)
JP (2) JP6069755B2 (fr)
CN (1) CN102272153B (fr)
AU (1) AU2009317161B2 (fr)
CA (1) CA2743669C (fr)
DK (1) DK2352756T3 (fr)
ES (1) ES2394180T3 (fr)
WO (1) WO2010058023A1 (fr)

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8697854B2 (en) 2008-11-24 2014-04-15 Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt Gmbh High affinity T cell receptor and use thereof
US9181527B2 (en) 2009-10-29 2015-11-10 The Trustees Of Dartmouth College T cell receptor-deficient T cell compositions
US9238063B2 (en) 2001-03-16 2016-01-19 Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundlheit und Umwelt (GmbH) Semi-allogenic anti-tumour vaccine with HLA haplo-identical antigen-presenting cells
US9273283B2 (en) 2009-10-29 2016-03-01 The Trustees Of Dartmouth College Method of producing T cell receptor-deficient T cells expressing a chimeric receptor
AU2016202021B2 (en) * 2009-02-09 2017-09-07 Helmholtz Zentrum München Deutsches Forschungszentrum Fuer Gesundheit Und Umwelt (Gmbh) Repertoire of allo-restricted peptide-specific t cell receptor sequences and use thereof
US9790278B2 (en) 2012-05-07 2017-10-17 The Trustees Of Dartmouth College Anti-B7-H6 antibody, fusion proteins, and methods of using the same
US9833476B2 (en) 2011-08-31 2017-12-05 The Trustees Of Dartmouth College NKP30 receptor targeted therapeutics
US10336804B2 (en) 2004-09-24 2019-07-02 Trustees Of Dartmouth College Chimeric NK receptor and methods for treating cancer
US10364237B2 (en) 2014-05-05 2019-07-30 Lycera Corporation Tetrahydroquinoline sulfonamide and related compounds for use as agonists of RORγ and the treatment of disease
US10421751B2 (en) 2015-05-05 2019-09-24 Lycera Corporation Dihydro-2H-benzo[b][1,4]oxazine sulfonamide and related compounds for use as agonists of RORγ and the treatment of disease
US10532088B2 (en) 2014-02-27 2020-01-14 Lycera Corporation Adoptive cellular therapy using an agonist of retinoic acid receptor-related orphan receptor gamma and related therapeutic methods
US10538572B2 (en) 2014-08-04 2020-01-21 Fred Hutchinson Cancer Research Center T cell immunotherapy specific for WT-1
US10611740B2 (en) 2015-06-11 2020-04-07 Lycera Corporation Aryl dihydro-2H-benzo[b][1,4]oxazine sulfonamide and related compounds for use as agonists of RORγ and the treatment of disease
US10875904B2 (en) 2012-05-03 2020-12-29 Fred Hutchinson Cancer Research Center Enhanced affinity T cell receptors and methods for making the same
WO2021198163A1 (fr) 2020-04-01 2021-10-07 Medigene Immunotherapies Gmbh Protéine de fusion cd3 et ses utilisations
US11596654B2 (en) 2016-06-10 2023-03-07 Gadeta B.V. Human leukocyte antigen restricted gamma delta T cell receptors and methods of use thereof
US11686724B2 (en) 2012-03-28 2023-06-27 Gadeta B.V. Compositions comprising gamma 9 delta 2 T-cell receptors and methods of use thereof to treat cancer

Families Citing this family (87)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101330830B (zh) 2005-10-18 2016-01-20 国家犹太健康中心 条件无限增殖化长期干细胞和制备和使用所述细胞的方法
EP2285832B1 (fr) 2008-05-16 2020-08-26 Taiga Biotechnologies, Inc. Anticorps et leurs procédés de préparation
DK2966084T3 (en) 2008-08-28 2018-08-06 Taiga Biotechnologies Inc MODULATORS OF MYC, PROCEDURES FOR USING SAME AND PROCEDURES FOR IDENTIFYING SUBSTANCES MODULATING MYC
ES2856179T3 (es) 2012-07-20 2021-09-27 Taiga Biotechnologies Inc Reconstitución y autorreconstitución potenciadas del compartimento hematopoyético
US10272115B2 (en) 2013-03-11 2019-04-30 Taiga Biotechnologies, Inc. Production and use of red blood cells
GB201309421D0 (en) * 2013-05-24 2013-07-10 Imp Innovations Ltd Polypeptides
RU2645256C2 (ru) * 2013-06-26 2018-02-19 Гуандун Сянсюэ Лайф Сайенсис, Лтд. Высокостабильный т-клеточный рецептор и способ его получения и применения
US10023625B2 (en) * 2013-11-22 2018-07-17 The Board Of Trustees Of The University Of Illinois Engineered high-affinity human T cell receptors
US10801070B2 (en) 2013-11-25 2020-10-13 The Broad Institute, Inc. Compositions and methods for diagnosing, evaluating and treating cancer
US11725237B2 (en) 2013-12-05 2023-08-15 The Broad Institute Inc. Polymorphic gene typing and somatic change detection using sequencing data
KR20230076867A (ko) 2013-12-20 2023-05-31 더 브로드 인스티튜트, 인코퍼레이티드 신생항원 백신과의 병용 요법
EP3099708B1 (fr) * 2014-01-29 2019-09-04 University Health Network Procédés et compositions permettant la production d'une cellule exprimant un récepteur de l'antigène des lymphocytes t
AU2015338958A1 (en) * 2014-10-31 2017-05-11 Baylor College Of Medicine Survivin specific T-cell receptor targeting tumor but not T cells
WO2016071758A1 (fr) * 2014-11-03 2016-05-12 Leiden University Medical Center Récepteurs de cellules t dirigées contre bob1 et leurs utilisations
EP3234130B1 (fr) 2014-12-19 2020-11-25 The Broad Institute, Inc. Procédé pour profiler le répertoire de récepteurs de cellulles t.
US10975442B2 (en) 2014-12-19 2021-04-13 Massachusetts Institute Of Technology Molecular biomarkers for cancer immunotherapy
US11680244B2 (en) 2015-05-20 2023-06-20 The Regents Of The University Of California Method for generating human dendritic cells for immunotherapy
AU2016264623B2 (en) 2015-05-20 2022-06-30 Dana-Farber Cancer Institute, Inc. Shared neoantigens
AU2016267155A1 (en) * 2015-05-26 2017-12-21 Advaxis, Inc. Personalized delivery vector-based immunotherapy and uses thereof
CA3012607A1 (fr) 2015-06-18 2016-12-22 The Broad Institute Inc. Enzymes et systemes crispr
AU2016317936A1 (en) * 2015-09-04 2018-03-08 Tocagen Inc. Recombinant vectors comprising 2A peptide
US20190255107A1 (en) 2015-10-09 2019-08-22 The Brigham And Women's Hospital, Inc. Modulation of novel immune checkpoint targets
EP3368689B1 (fr) 2015-10-28 2020-06-17 The Broad Institute, Inc. Compositions d'évaluation et de modulation des réponses immunitaires à l'aide de signatures génétiques de cellules immunitaires
WO2017075451A1 (fr) 2015-10-28 2017-05-04 The Broad Institute Inc. Compositions et procédés d'évaluation et de modulation des réponses immunitaires par détection et ciblage de pou2af1
WO2017075465A1 (fr) 2015-10-28 2017-05-04 The Broad Institute Inc. Compositions et procédés d'évaluation et de modulation des réponses immunitaires par détection et ciblage de gata3
US11154573B2 (en) * 2015-10-30 2021-10-26 The Regents Of The University Of California Methods of generating T-cells from stem cells and immunotherapeutic methods using the T-cells
AU2016355178B9 (en) 2015-11-19 2019-05-30 Massachusetts Institute Of Technology Lymphocyte antigen CD5-like (CD5L)-interleukin 12B (p40) heterodimers in immunity
WO2017139405A1 (fr) * 2016-02-08 2017-08-17 The General Hospital Corporation Procédé permettant de réduire la capacité de tolérance de lymphocytes t par rapport à des antigènes tumoraux
CN109641008B (zh) 2016-04-01 2021-03-23 凯德药业股份有限公司 嵌合受体及其方法和用途
CU20180121A7 (es) 2016-04-01 2019-05-03 Kite Pharma Inc Receptores de antígenos quiméricos y células t
CA3019135A1 (fr) 2016-04-01 2017-10-05 Kite Pharma, Inc. Molecules de liaison a bcma et procedes d'utilisation associes
EP3443123B1 (fr) * 2016-04-11 2021-08-11 Board of Regents, The University of Texas System Procédés et compositions pour détecter la séquence et l'affinité vis-à-vis d'un récepteur unique de lymphocyte t
AU2017254477A1 (en) 2016-04-18 2018-11-01 Jennifer G. ABELIN Improved HLA epitope prediction
US11630103B2 (en) 2016-08-17 2023-04-18 The Broad Institute, Inc. Product and methods useful for modulating and evaluating immune responses
US20190262399A1 (en) 2016-09-07 2019-08-29 The Broad Institute, Inc. Compositions and methods for evaluating and modulating immune responses
US20200016202A1 (en) 2016-10-07 2020-01-16 The Brigham And Women's Hospital, Inc. Modulation of novel immune checkpoint targets
AU2017367730A1 (en) 2016-12-02 2019-06-06 Taiga Biotechnologies, Inc. Nanoparticle formulations
KR102375218B1 (ko) * 2016-12-08 2022-03-17 이매틱스 바이오테크놀로지스 게엠베하 T 세포 수용체 및 이를 사용하는 면역 요법
EP3565565A4 (fr) 2017-01-04 2020-12-09 Nova Southeastern University Cellules tueuses naturelles (nk) exprimant un complexe récepteur des lymphocytes t fonctionnels spécifiques d'un antigène (tcr), leurs méthodes de production et leurs méthodes d'utilisation thérapeutique
US11549149B2 (en) 2017-01-24 2023-01-10 The Broad Institute, Inc. Compositions and methods for detecting a mutant variant of a polynucleotide
HUE065174T2 (hu) 2017-02-12 2024-05-28 Biontech Us Inc HLA-alapú módszerek és készítmények, valamint azok felhasználása
US11963966B2 (en) 2017-03-31 2024-04-23 Dana-Farber Cancer Institute, Inc. Compositions and methods for treating ovarian tumors
US11913075B2 (en) 2017-04-01 2024-02-27 The Broad Institute, Inc. Methods and compositions for detecting and modulating an immunotherapy resistance gene signature in cancer
EP3610266A4 (fr) 2017-04-12 2021-04-21 Massachusetts Eye and Ear Infirmary Signature tumorale pour métastase, compositions de matière et leurs procédés d'utilisation
WO2018195019A1 (fr) 2017-04-18 2018-10-25 The Broad Institute Inc. Composition permettant de détecter une sécrétion et procédé d'utilisation
EP3638218A4 (fr) 2017-06-14 2021-06-09 The Broad Institute, Inc. Compositions et procédés ciblant le composant 3 du complément pour inhiber la croissance tumorale
AU2017425657A1 (en) 2017-08-03 2020-02-13 Taiga Biotechnologies, Inc. Methods and compositions for the treatment of melanoma
US10149898B2 (en) 2017-08-03 2018-12-11 Taiga Biotechnologies, Inc. Methods and compositions for the treatment of melanoma
JP2020532323A (ja) * 2017-09-05 2020-11-12 グリットストーン オンコロジー インコーポレイテッド Ｔ細胞療法用の新生抗原の特定法
CN111511388A (zh) 2017-09-21 2020-08-07 博德研究所 用于靶向核酸编辑的系统、方法和组合物
CA3078472A1 (fr) 2017-10-06 2019-04-11 Oslo Universitetssykehus Hf Recepteurs d'antigenes chimeriques
WO2019084055A1 (fr) 2017-10-23 2019-05-02 Massachusetts Institute Of Technology Classification de variation génétique à partir de transcriptomes unicellulaires
WO2019094983A1 (fr) 2017-11-13 2019-05-16 The Broad Institute, Inc. Méthodes et compositions de traitement du cancer par ciblage de la voie clec2d-klrb1
CN111630602A (zh) 2017-11-22 2020-09-04 磨石肿瘤生物技术公司 减少新抗原的接合表位呈递
EP3714041A1 (fr) 2017-11-22 2020-09-30 Iovance Biotherapeutics, Inc. Expansion de lymphocytes de sang périphérique (pbl) à partir de sang périphérique
US11994512B2 (en) 2018-01-04 2024-05-28 Massachusetts Institute Of Technology Single-cell genomic methods to generate ex vivo cell systems that recapitulate in vivo biology with improved fidelity
US11957695B2 (en) 2018-04-26 2024-04-16 The Broad Institute, Inc. Methods and compositions targeting glucocorticoid signaling for modulating immune responses
WO2019232542A2 (fr) 2018-06-01 2019-12-05 Massachusetts Institute Of Technology Procédés et compositions pour détecter et moduler des signatures géniques micro-environnementales à partir du lcr de patients présentant des métastases
WO2019241315A1 (fr) 2018-06-12 2019-12-19 Obsidian Therapeutics, Inc. Constructions régulatrices dérivées de pde5 et procédés d'utilisation en immunothérapie
US20210355522A1 (en) 2018-08-20 2021-11-18 The Broad Institute, Inc. Inhibitors of rna-guided nuclease activity and uses thereof
WO2020068304A2 (fr) 2018-08-20 2020-04-02 The Broad Institute, Inc. Inhibiteurs de liaison de cible de nucléase guidée par arn et leurs utilisations
WO2020041387A1 (fr) 2018-08-20 2020-02-27 The Brigham And Women's Hospital, Inc. Modifications de domaine de dégradation pour la régulation spatio-temporelle de nucléases guidées par arn
US20210382068A1 (en) 2018-10-02 2021-12-09 Dana-Farber Cancer Institute, Inc. Hla single allele lines
WO2020081730A2 (fr) 2018-10-16 2020-04-23 Massachusetts Institute Of Technology Méthodes et compositions pour moduler un microenvironnement
WO2020092455A2 (fr) 2018-10-29 2020-05-07 The Broad Institute, Inc. Atlas de transcription de lymphocytes car-t
US20220062394A1 (en) 2018-12-17 2022-03-03 The Broad Institute, Inc. Methods for identifying neoantigens
CN113302491A (zh) * 2018-12-18 2021-08-24 Mbl国际公司 pMHC占有率的链霉亲和素-寡核苷酸缀合物的组成
US11739156B2 (en) 2019-01-06 2023-08-29 The Broad Institute, Inc. Massachusetts Institute of Technology Methods and compositions for overcoming immunosuppression
MX2021010288A (es) 2019-03-01 2021-09-23 Iovance Biotherapeutics Inc Expansion de linfocitos infiltrantes de tumores a partir de tumores liquidos y usos terapeuticos de los mismos.
WO2020186101A1 (fr) 2019-03-12 2020-09-17 The Broad Institute, Inc. Procédés de détection, compositions et méthodes de modulation des cellules de sarcome synovial
EP3942023A1 (fr) 2019-03-18 2022-01-26 The Broad Institute, Inc. Compositions et méthodes de modulation de régulateurs métaboliques de pathogénicité de lymphocytes t
US20220235340A1 (en) 2019-05-20 2022-07-28 The Broad Institute, Inc. Novel crispr-cas systems and uses thereof
US20220226464A1 (en) 2019-05-28 2022-07-21 Massachusetts Institute Of Technology Methods and compositions for modulating immune responses
CN110357953B (zh) * 2019-07-17 2022-05-03 深圳市因诺转化医学研究院 识别人巨细胞病毒pp65抗原的TCR
WO2021030627A1 (fr) 2019-08-13 2021-02-18 The General Hospital Corporation Procédés de prédiction de résultats d'inhibition de point de contrôle et traitement associés
WO2021041922A1 (fr) 2019-08-30 2021-03-04 The Broad Institute, Inc. Systèmes de transposase mu associés à crispr
US11981922B2 (en) 2019-10-03 2024-05-14 Dana-Farber Cancer Institute, Inc. Methods and compositions for the modulation of cell interactions and signaling in the tumor microenvironment
US11793787B2 (en) 2019-10-07 2023-10-24 The Broad Institute, Inc. Methods and compositions for enhancing anti-tumor immunity by targeting steroidogenesis
US11844800B2 (en) 2019-10-30 2023-12-19 Massachusetts Institute Of Technology Methods and compositions for predicting and preventing relapse of acute lymphoblastic leukemia
US11865168B2 (en) 2019-12-30 2024-01-09 Massachusetts Institute Of Technology Compositions and methods for treating bacterial infections
WO2021185775A1 (fr) * 2020-03-16 2021-09-23 Biontech Cell & Gene Therapies Gmbh Récepteurs de lymphocytes t spécifiques à un antigène et épitopes de lymphocytes t
US20240150711A1 (en) 2021-03-01 2024-05-09 Dana-Farber Cancer Institute, Inc. Personalized redirection and reprogramming of t cells for precise targeting of tumors
AU2022288058A1 (en) 2021-06-07 2023-11-16 Agonox, Inc. Cxcr5, pd-1, and icos expressing tumor reactive cd4 t cells and their use
CN113929767B (zh) * 2021-10-14 2023-11-24 深圳大学总医院 具有高亲和力的t细胞受体及其用途
WO2024077256A1 (fr) 2022-10-07 2024-04-11 The General Hospital Corporation Procédés et compositions pour la découverte à haut débit de protéines de liaison ciblant un peptide-cmh
WO2024098024A1 (fr) 2022-11-04 2024-05-10 Iovance Biotherapeutics, Inc. Expansion de lymphocytes infiltrant les tumeurs à partir de tumeurs liquides et leurs utilisations thérapeutiques
WO2024124044A1 (fr) 2022-12-07 2024-06-13 The Brigham And Women’S Hospital, Inc. Compositions et procédés ciblant sat1 pour améliorer l'immunité antitumorale pendant la progression d'une tumeur

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1870418A1 (fr) * 2006-06-20 2007-12-26 GSF-Forschungszentrum für Umwelt und Gesundheit GmbH Allolimitée peptide spécifiques pour les cellules T

Family Cites Families (47)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4415553A (en) 1973-08-06 1983-11-15 Dso "Pharmachim" Compositions, processes for their preparation and method for treatment of neoplasms
US4568542A (en) 1981-06-09 1986-02-04 Lee Biomolecular Research Laboratories, Inc. Vaccine compositions
JPH0751511B2 (ja) 1982-03-15 1995-06-05 味の素株式会社 インターロイキン２を含有してなる癌治療剤
US4703004A (en) 1984-01-24 1987-10-27 Immunex Corporation Synthesis of protein with an identification peptide
US4877611A (en) 1986-04-15 1989-10-31 Ribi Immunochem Research Inc. Vaccine containing tumor antigens and adjuvants
US4851341A (en) 1986-12-19 1989-07-25 Immunex Corporation Immunoaffinity purification system
US5906936A (en) 1988-05-04 1999-05-25 Yeda Research And Development Co. Ltd. Endowing lymphocytes with antibody specificity
US5582831A (en) 1991-11-26 1996-12-10 Yeda Research And Development Co., Ltd. Anti-tumor vaccines
US5156841A (en) 1988-08-26 1992-10-20 United States Of America Anti-tumor vaccine
US5290551A (en) 1990-05-08 1994-03-01 Thomas Jefferson University Treatment of melanoma with a vaccine comprising irradiated autologous melanoma tumor cells conjugated to a hapten
US6077519A (en) 1993-01-29 2000-06-20 University Of Pittsburgh Methods for isolation and use of T cell epitopes eluted from viable cells in vaccines for treating cancer patients
EP0997477B1 (fr) 1994-07-29 2006-03-15 Sunol Molecular Corporation MHC-complexes et leurs applications
US6143292A (en) 1995-05-25 2000-11-07 Baxter International Inc. Allogeneic cell therapy for cancer following allogeneic stem cell transplantation
US5788963A (en) 1995-07-31 1998-08-04 Pacific Northwest Cancer Foundation Isolation and/or preservation of dendritic cells for prostate cancer immunotherapy
US6805861B2 (en) 1996-01-17 2004-10-19 Imperial College Innovations Limited Immunotherapy using cytotoxic T lymphocytes (CTL)
CA2246333A1 (fr) * 1996-03-05 1997-09-12 The Scripps Research Institute Elements recombinants codant pour des recepteurs de lymphocytes t specifiques des antigenes de tumeurs limites par hla humains
US5853719A (en) 1996-04-30 1998-12-29 Duke University Methods for treating cancers and pathogen infections using antigen-presenting cells loaded with RNA
US6458585B1 (en) 1996-08-14 2002-10-01 Nexell Therapeutics Inc. Cytokine-free culture of dendritic cells
US6063375A (en) 1996-09-10 2000-05-16 Medical University Of South Carolina Semiallogeneic cell hybrids and related methods for treating cancer
CA2278678A1 (fr) 1997-01-31 1998-08-06 Research Corporation Technologies, Inc. Immunotherapie anticancer a base de cellules semi-allogeniques
AU7983198A (en) 1997-06-23 1999-01-04 Ludwig Institute For Cancer Research Improved methods for inducing an immune response
CN1270630A (zh) 1997-07-21 2000-10-18 塞鲁斯公司 处理白细胞的方法、白细胞组合物及其应用方法
WO2000076537A2 (fr) 1997-09-10 2000-12-21 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Utilisation de complexes de peptide-lignee cellulaire semi-allogenique§ pour le traitement du cancer, du sida et d'autres maladies virales
AU1601801A (en) 1999-11-16 2001-05-30 Eric M. Eastman Methods of preparing crna
US20020123479A1 (en) 1999-12-08 2002-09-05 Song Elizabeth S. Immunostimulation mediated by gene-modified dendritic cells
US20020168351A1 (en) 2000-10-20 2002-11-14 Tsuneya Ohno Fusion cells and cytokine compositions for treatment of disease
EP1201760A1 (fr) 2000-10-30 2002-05-02 ARTEMIS Pharmaceuticals GmbH Vecteur viral Influenza pour cellules dendritiques
AU2002237217A1 (en) 2001-03-12 2002-09-24 Cellcure Aps Continuous, normal human t-lymphocyte cell lines comprising a recombinant immune receptor with defined antigen specificity
DE10112851C1 (de) 2001-03-16 2002-10-10 Gsf Forschungszentrum Umwelt Semi-allogene Antitumor-Vakzine mit HLA-haplo-identischen Antigen-präsentierenden Zellen
DE10132502A1 (de) 2001-07-05 2003-01-23 Gsf Forschungszentrum Umwelt Angriff auf Tumorzellen mit fehlender, niedriger oder anormaler MHC-Expression durch kombinieren von nicht MHC-Restringierten T-Zellen/NK-Zellen und MHC-Restringierten Zellen
EP1420253B1 (fr) 2002-08-23 2006-05-31 Deutsches Rheuma-Forschungszentrum Berlin Procédé de détection et isolation des lymphocytes T reconnaissant un antigène defini
US7915036B2 (en) 2004-09-13 2011-03-29 The United States Of America As Represented By The Secretary, Department Of Health And Human Services Compositions comprising T cell receptors and methods of use thereof
WO2006065495A2 (fr) 2004-11-24 2006-06-22 Fred Hutchinson Cancer Research Center Methodes d'utilisation d'il-21 pour l'immunotherapie adoptive et l'identification d'antigenes tumoraux
PT2327763T (pt) 2005-08-05 2018-05-11 Helmholtz Zentrum Muenchen Deutsches Forschungszentrum Gesundheit & Umwelt Gmbh Geração de células t específicas de antigénios
EP1795599A1 (fr) 2005-12-09 2007-06-13 Schuler, Gerold, Prof. Dr. Procédé pour la préparation de cellules T effectrices
WO2007131092A2 (fr) 2006-05-03 2007-11-15 Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Récepteurs de lymphocytes t chimériques, matières associées et méthodes d'utilisation
EP1878744A1 (fr) 2006-07-13 2008-01-16 Max-Delbrück-Centrum für Molekulare Medizin (MDC) Epitope-tag pour des récepteurs de cellules T exprimés en surface, leurs utilisations et méthode de selection de cellules hôtes les exprimant
WO2008039818A2 (fr) 2006-09-26 2008-04-03 Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Récepteurs des cellules t modifiés, et matériaux et méthodes s'y rapportant
EP1932537A1 (fr) 2006-12-12 2008-06-18 Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) l'expression de recepteurs des lymphocytes t transgeniques dans lymphocytes lak-t
JP5292550B2 (ja) * 2007-03-23 2013-09-18 静岡県 Ｔ細胞レセプターβ鎖遺伝子及びα鎖遺伝子
AU2008234530B2 (en) * 2007-03-29 2013-03-28 Technion Research & Development Foundation Ltd. Antibodies, methods and kits for diagnosing and treating melanoma
US20110280894A1 (en) 2008-07-31 2011-11-17 Angela Krackhardt Her2/neu specific t cell receptors
US8697854B2 (en) 2008-11-24 2014-04-15 Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt Gmbh High affinity T cell receptor and use thereof
WO2010089412A1 (fr) * 2009-02-09 2010-08-12 Helmholtz Zentrum Muenchen Deutsches Forschungszentrum Fuer Gesundheit Und Umwelt (Gmbh) Répertoire immunologique de séquences de récepteurs de lymphocytes t spécifiques de peptides allo-restreints et son utilisation
AU2015338958A1 (en) * 2014-10-31 2017-05-11 Baylor College Of Medicine Survivin specific T-cell receptor targeting tumor but not T cells
CA2985029A1 (fr) * 2015-05-04 2016-11-10 Vcn Biosciences Sl Adenovirus oncolytiques comprenant des mutations dans les epitopes d'adenovirus immunodominants et leur utilisation dans le traitement d'un cancer
JP2023519727A (ja) * 2020-04-01 2023-05-12 メディジーン イミュノテラピーズ ゲーエムベーハー Ｃｄ３融合タンパク質およびその使用

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1870418A1 (fr) * 2006-06-20 2007-12-26 GSF-Forschungszentrum für Umwelt und Gesundheit GmbH Allolimitée peptide spécifiques pour les cellules T

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
AMROLIA PERSIS J ET AL: "Allorestricted cytotoxic T cells specific for human CD45 show potent antileukemic activity.", BLOOD, vol. 101, no. 3, 1 February 2003 (2003-02-01), pages 1007 - 1014, XP002523562, ISSN: 0006-4971 *
KAZANSKY DMITRY B: "Intrathymic selection: New insight into tumor immunology", ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY SPRINGER-VERLAG BERLIN, HEIDELBERGER PLATZ 3, D-14197 BERLIN, GERMANY SERIES : ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY (ISSN 0065-2598(PRINT)), 2007, & INTERNATIONAL IMMUNE-MEDIATED DISEASES CONGRESS; MOSCOW, RUSSIA; OCTOBER 03 -08, 2005, pages 133 - 144, XP002523561, ISSN: 978-0-387-72004-3(H) *
STAUSS H J: "Immunotherapy with CTLs restricted by nonself MHC", IMMUNOLOGY TODAY, ELSEVIER PUBLICATIONS, CAMBRIDGE, GB, vol. 20, no. 4, 1 April 1999 (1999-04-01), pages 180 - 183, XP004162795, ISSN: 0167-5699 *
VISSEREN MARJAN J W ET AL: "Affinity, specificity and T-cell-receptor diversity of melanoma-specific CTL generated in vitro against a single tyrosinase epitope", INTERNATIONAL JOURNAL OF CANCER, vol. 72, no. 6, 1997, pages 1122 - 1128, XP002523563, ISSN: 0020-7136 *

Cited By (38)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9238063B2 (en) 2001-03-16 2016-01-19 Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundlheit und Umwelt (GmbH) Semi-allogenic anti-tumour vaccine with HLA haplo-identical antigen-presenting cells
US9597384B2 (en) 2001-03-16 2017-03-21 Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt (Gmbh) Semi-allogenic anti-tumour vaccine with HLA haplo-identical antigen-presenting cells
US11208454B2 (en) 2004-09-24 2021-12-28 Trustees Of Dartmouth College Chimeric NK receptor and methods for treating cancer
US11858976B2 (en) 2004-09-24 2024-01-02 The Trustees Of Dartmouth College Nucleic acid constructs encoding chimeric NK receptor, cells containing, and therapeutic use thereof
US10336804B2 (en) 2004-09-24 2019-07-02 Trustees Of Dartmouth College Chimeric NK receptor and methods for treating cancer
US9862755B2 (en) 2008-11-24 2018-01-09 Max-Delbrueck-Centrum Fuer Molekulare Medizin High affinity T cell receptor and use thereof
US10626159B2 (en) 2008-11-24 2020-04-21 Max-Delbrueck-Centrum Fuer Molekulare Medizin High affinity T cell receptor and use thereof
US8697854B2 (en) 2008-11-24 2014-04-15 Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt Gmbh High affinity T cell receptor and use thereof
AU2016202021B2 (en) * 2009-02-09 2017-09-07 Helmholtz Zentrum München Deutsches Forschungszentrum Fuer Gesundheit Und Umwelt (Gmbh) Repertoire of allo-restricted peptide-specific t cell receptor sequences and use thereof
US9181527B2 (en) 2009-10-29 2015-11-10 The Trustees Of Dartmouth College T cell receptor-deficient T cell compositions
US10689617B1 (en) 2009-10-29 2020-06-23 The Trustees Of Dartmouth College T-cell receptor-deficient T cell compositions
US10689619B2 (en) 2009-10-29 2020-06-23 The Trustees Of Dartmouth College T cell receptor-deficient T cell compositions
US9938497B2 (en) 2009-10-29 2018-04-10 The Trustees Of Dartmouth College T cell receptor-deficient T cell compositions
US9663763B2 (en) 2009-10-29 2017-05-30 The Trustees Of Dartmouth College T-cell receptor-deficient T cell compositions
US10689618B2 (en) 2009-10-29 2020-06-23 The Trustees Of Dartmouth College T cell receptor-deficient T cell compositions
US10689616B1 (en) 2009-10-29 2020-06-23 The Trustees Of Dartmouth College T-cell receptor-deficient t cell compositions
US11136549B2 (en) 2009-10-29 2021-10-05 The Trustees Of Dartmouth College T-cell receptor-deficient T cell compositions
US9822340B1 (en) 2009-10-29 2017-11-21 The Trustees Of Dartmouth College T-cell receptor-deficient T cell compositions
US9957480B2 (en) 2009-10-29 2018-05-01 The Trustees Of Dartmouth College T cell receptor-deficient T cell compositions
US11834676B2 (en) 2009-10-29 2023-12-05 The Trustees Of Dartmouth College T cell receptor-deficient T cell compositions
US9273283B2 (en) 2009-10-29 2016-03-01 The Trustees Of Dartmouth College Method of producing T cell receptor-deficient T cells expressing a chimeric receptor
US9821011B1 (en) 2009-10-29 2017-11-21 The Trustees Of Dartmouth College T-cell receptor-deficient T cell compositions
US11872248B2 (en) 2011-08-31 2024-01-16 The Trustees Of Dartmouth College Nucleic acids encoding chimeric receptor comprising NKP30 receptor and CD28 and CD3 zeta domains and human T cell containing
US10682378B2 (en) 2011-08-31 2020-06-16 The Trustees Of Dartmouth College NKP30 receptor targeted therapeutics
US9833476B2 (en) 2011-08-31 2017-12-05 The Trustees Of Dartmouth College NKP30 receptor targeted therapeutics
US11686724B2 (en) 2012-03-28 2023-06-27 Gadeta B.V. Compositions comprising gamma 9 delta 2 T-cell receptors and methods of use thereof to treat cancer
US10875904B2 (en) 2012-05-03 2020-12-29 Fred Hutchinson Cancer Research Center Enhanced affinity T cell receptors and methods for making the same
US11034766B2 (en) 2012-05-07 2021-06-15 Trustees Of Dartmouth College Anti-B7-H6 antibody, fusion proteins, and methods of using the same
US9790278B2 (en) 2012-05-07 2017-10-17 The Trustees Of Dartmouth College Anti-B7-H6 antibody, fusion proteins, and methods of using the same
US10532088B2 (en) 2014-02-27 2020-01-14 Lycera Corporation Adoptive cellular therapy using an agonist of retinoic acid receptor-related orphan receptor gamma and related therapeutic methods
US10442798B2 (en) 2014-05-05 2019-10-15 Lycera Corporation Tetrahydroquinoline sulfonamide and related compounds for use as agonists of RORγ and the treatment of disease
US10364237B2 (en) 2014-05-05 2019-07-30 Lycera Corporation Tetrahydroquinoline sulfonamide and related compounds for use as agonists of RORγ and the treatment of disease
US10538572B2 (en) 2014-08-04 2020-01-21 Fred Hutchinson Cancer Research Center T cell immunotherapy specific for WT-1
US10421751B2 (en) 2015-05-05 2019-09-24 Lycera Corporation Dihydro-2H-benzo[b][1,4]oxazine sulfonamide and related compounds for use as agonists of RORγ and the treatment of disease
US10611740B2 (en) 2015-06-11 2020-04-07 Lycera Corporation Aryl dihydro-2H-benzo[b][1,4]oxazine sulfonamide and related compounds for use as agonists of RORγ and the treatment of disease
US11059796B2 (en) 2015-06-11 2021-07-13 The Regents Of The University Of Michigan Aryl dihydro-2H benzo[b][1,4]oxazine sulfonamide and related compounds for use as agonists of RORγ and the treatment of disease
US11596654B2 (en) 2016-06-10 2023-03-07 Gadeta B.V. Human leukocyte antigen restricted gamma delta T cell receptors and methods of use thereof
WO2021198163A1 (fr) 2020-04-01 2021-10-07 Medigene Immunotherapies Gmbh Protéine de fusion cd3 et ses utilisations

Also Published As

Publication number Publication date
AU2009317161B2 (en) 2014-09-11
CA2743669A1 (fr) 2010-05-27
US20200339653A1 (en) 2020-10-29
EP2352756A1 (fr) 2011-08-10
US20110280889A1 (en) 2011-11-17
US8697854B2 (en) 2014-04-15
JP6069755B2 (ja) 2017-02-01
US20180162921A1 (en) 2018-06-14
AU2009317161A1 (en) 2010-05-27
US20140348805A1 (en) 2014-11-27
CN102272153B (zh) 2015-04-15
CA2743669C (fr) 2018-10-16
ES2394180T3 (es) 2013-01-23
EP2352756B1 (fr) 2012-09-19
CN102272153A (zh) 2011-12-07
US9862755B2 (en) 2018-01-09
US10626159B2 (en) 2020-04-21
JP2012509659A (ja) 2012-04-26
JP2015211682A (ja) 2015-11-26
DK2352756T3 (da) 2012-12-03

Similar Documents

Publication Publication Date Title
US20200339653A1 (en) High affinity t cell receptor and use thereof
AU2015230741B2 (en) Repertoire of allo-restricted peptide-specific t cell receptor sequences and use thereof
EP2041171B1 (fr) Etiquette epitopique pour des proteines de surface et leurs utilisations
Wilde et al. Dendritic cells pulsed with RNA encoding allogeneic MHC and antigen induce T cells with superior antitumor activity and higher TCR functional avidity
US20190010207A1 (en) Pd-1-cd28 fusion proteins and their use in medicine
JP2022133308A (ja) キメラアロ抗原受容体ｔ細胞の組成物および方法
KR20190016507A (ko) TGFβRII의 프레임시프트 돌연변이체를 인식하는 T-세포 수용체
US20200308279A1 (en) Chimeric antigen receptors
WO2022098750A1 (fr) Tcr restreints au hla de classe ii contre la mutation activant kras g12>v
GB2569692A (en) T cell antigen receptor chimera
EA042909B1 (ru) Т-КЛЕТОЧНЫЕ РЕЦЕПТОРЫ, КОТОРЫЕ РАСПОЗНАЮТ МУТАНТНЫЕ ВАРИАНТЫ СО СДВИГОМ РАМКИ СЧИТЫВАНИЯ TGFβRII
Schendel Dendritic cells pulsed with RNA encoding allogeneic MHC and antigen induce T cells with superior anti-tumor activity and higher TCR functional avidity

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 200980154272.4

Country of ref document: CN

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 09760145

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2009760145

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2009317161

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 2743669

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 2011536895

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2009317161

Country of ref document: AU

Date of ref document: 20091124

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 13130665

Country of ref document: US