CN115698038A - Signal transduction receptors and uses thereof - Google Patents

Signal transduction receptors and uses thereof Download PDF

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CN115698038A
CN115698038A CN202180039676.XA CN202180039676A CN115698038A CN 115698038 A CN115698038 A CN 115698038A CN 202180039676 A CN202180039676 A CN 202180039676A CN 115698038 A CN115698038 A CN 115698038A
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金华君
王超
郭晓春
黄晨
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Shanghai Junsai Biotechnology Co ltd
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Abstract

Signal-transducing receptors and uses thereof are provided. The signal transduction receptor comprises the extracellular domain of a receptor for immunosuppressive cytokines or a functional fragment or mutant thereof retaining specific binding to immunosuppressive cytokines; a transmembrane region or a mutant thereof; and the intracellular domain of the costimulatory signaling molecule or a functional fragment or mutant thereof that retains the biological function of the costimulatory signaling molecule to deliver a costimulatory signal to activate an immune cell. After the immune cells modified by the signal conversion receptor contact molecules with the surface of tumor cells and/or the surface of tumor stromal cells to play a role in immunosuppression, the immune cells are not influenced by the negative effects of immunosuppressive molecules, but play a role in immune activation, enhance the proliferation capacity of the immune cells and the secretion function of cytokines, prolong the survival time of the activated immune cells and further overcome the side effects of tumor immunosuppressive microenvironment on tumor adoptive cell therapy effector cells.

Description

Signal transduction receptors and uses thereof
The present application claims priority to the invention patent application entitled "signal transduction receptor and uses thereof" filed 6/1/2020, china application No. 202010485006.8, the entire contents of which are incorporated herein by reference.
Technical Field
The invention relates to the field of biotechnology, in particular to a signal conversion receptor and application thereof.
Background
In Adoptive Cell Therapy (ACT) of tumors, the stability of the immune system is of crucial importance. Too low a response may cause severe infection, and too strong a response may cause allergic reactions. The immune system of human body develops a fine and complex bidirectional immune regulation mechanism to carry out positive and negative bidirectional regulation on immune response. Activating and enhancing immune response when foreign antigenic foreign bodies are eliminated; after the foreign antigenic substance is eliminated, the immune response can be weakened to stop. In tumor ACT, positive regulatory molecules of effector T cells include CD3/TCR complex, CD28, CD134 (OX 40), CD137 (4-1 BB), etc., and negative regulatory molecules include PD-1 (PDCD 1, CD 279), CTLA-4 (CD 152), LAG3 (CD 223), TIM3 (HAVCR 2), etc. However, in the Tumor Microenvironment (TME), there are many factors that can negatively regulate the T cell immune response, allowing tumor cells to escape the monitoring and clearance of the body's immune system and continue to proliferate, invade and metastasize.
TGF- β has an inhibitory effect on the systemic immune system and inhibits immune surveillance by the host. TGF-. Beta.plays an important role in the initiation and progression of tumors. TGF- β has a tumor promoting effect, is commonly produced in large amounts in many types of tumors and is known to have carcinogenic effects, and has a negative effect on tumor immunity and significantly inhibits tumor immune monitoring by the host. TGF- β significantly and directly inhibits the cytotoxic program (cytotoxic program) of Cytotoxic T Lymphocytes (CTLs) by inhibiting the transcription of genes encoding a number of key proteins including at least: perforin, granzyme A, granzyme B, fas ligand and g interferon and other cytotoxins. At the same time, TGF-beta also has a significant effect on the differentiation and function of CD4+ T cells.
Aiming at an inhibitory signal pathway negatively regulated by immune effector cells, a plurality of reports have been reported in recent years to connect an extracellular region of a corresponding receptor with a transmembrane region and an intracellular region of a forward costimulatory signal molecule to construct a new fusion receptor, the fusion receptor plays a role of a signal converter after being expressed in the immune effector cells, and can convert immunosuppressive signals in a tumor microenvironment into immune activation signals, namely, the extracellular polypeptide of the signal conversion molecule receives signals of the immunosuppressive molecules on the surface of tumor cells, the surface of tumor stromal cells or in the tumor microenvironment and transmits the signals to the cells, and a second signal of the immune cells is activated by the intracellular segment of the costimulatory signal molecule, so that the proliferation and cytokine secretion functions of the immune effector cells are enhanced, and the survival time of the activated immune effector cells is prolonged. CN103965361B discloses a chimeric molecular converter of T cell signaling, which comprises the extracellular region of PD1 molecule (or fusion protein of PD1 extracellular region and HERIN), CD28 transmembrane region and CD28 (or 4-1 BB) intracellular costimulatory domain. The Tumor Infiltrating Lymphocyte (TIL) expressing the T cell signal chimeric molecule converter has obvious improvement on the proliferation effect and the killing effect on tumor target cells. CN105452287a discloses an immunosuppressive TGF- β signaling converter that is a chimera comprising an extracellular domain of a TGF- β receptor and an intracellular domain from another molecule, such that binding of TGF- β to the extracellular domain results in the intracellular domain stimulating T cell activity.
In order to stimulate the proliferation of immune effector cells and improve the tumor immunity, there is still a need for more means including signal transducer molecules to modify and activate immune effector cells.
Disclosure of Invention
The present invention provides an isolated fusion protein comprising: an extracellular domain of a receptor for an immunosuppressive cytokine or a functional fragment or mutant thereof that retains a specific binding immunosuppressive cytokine; a transmembrane region or a mutant thereof; and the intracellular domain of the costimulatory signaling molecule or a functional fragment or mutant thereof that retains the biological function of the costimulatory signaling molecule to deliver a costimulatory signal to activate an immune cell.
In one or more embodiments, the immunosuppressive cytokines include tumor cell or tumor stromal cell membrane proteins and tumor cell or tumor stromal cell secreted proteins.
In one or more embodiments, the tumor cell or tumor stromal cell membrane proteins include PDL1/PDL2, fasL, B7-H4, crry, and HLA-G.
In one or more embodiments, the tumor cell or tumor stromal cell secreted protein comprises IL13, IL4, TGF- β, IL6, IL8, IL-10, CCL21, IDO, and VEGF.
In one or more embodiments, the receptors for TGF- β include TGF- β receptor 1, TGF- β receptor 2, and TGF- β receptor 3, with TGF- β receptor 1 and TGF- β receptor 2 being preferred.
In one or more embodiments, the co-stimulatory signaling molecule comprises one or more of CD28, CD134 (OX 40), CD137 (4-1 BB), LCK, ICOS, DAP10, siglec-9, siglec-10, siglec-15, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, IL-R2, IL-4R, IL-7R, IL-10R, IL-12R, IL-15R, IL-21R, CD, and CD40.
In one or more embodiments, the transmembrane region comprises any one or more of the transmembrane regions from CD28, CD8, CD134 (OX 40), CD137 (4-1 BB), LCK, ICOS, DAP10, siglec-9, siglec-10, siglec-15, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, IL-2Rbeta, IL-2Rgamma, IL-4Ralpha, IL-7Ralpha, IL-10R, IL-12R, IL-15R, IL-21R, CD and CD40.
In one or more embodiments, the transmembrane region is from an IL-7Ralpha transmembrane region or a mutant thereof.
In one or more embodiments, the amino acid sequence of the mutant of the IL-7Ralpha transmembrane region is as shown in any one of SEQ ID NOS: 29-60.
In one or more embodiments, the coding sequence of the mutant of the IL-7Ralpha transmembrane region is as shown in any one of SEQ ID NOS 61-94.
In one or more embodiments, the extracellular region of the fusion protein is an extracellular region of TGF- β receptor 1, the transmembrane region is from an IL-7Ralpha transmembrane region or a mutant thereof, and the intracellular domain of the co-stimulatory molecule of the fusion protein is any one selected from the group consisting of an IL-7Ralpha intracellular region, a CD28 intracellular region in tandem 4-1BB intracellular region, a CD28 intracellular region, a 4-1BB intracellular region, a CD28 intracellular region in tandem OX40 intracellular region, a 28-IL2RB-z (YXXQ) intracellular region, a 28-DeltaIL2RB-z (YXXQ) intracellular region, an IL2RB-z (YXXQ) intracellular region, and a DeltaIL2RB-z (YXXQ) intracellular region.
In one or more embodiments, the amino acid sequence of the fusion protein is set forth in any one of SEQ ID NOs 95-350.
In one or more embodiments, the extracellular region of the fusion protein is an extracellular region of TGF- β receptor 2, the transmembrane region being from an IL-7Ralpha transmembrane region or a mutant thereof, and the intracellular domain of the co-stimulatory molecule of the fusion protein is any one selected from the group consisting of an IL-7Ralpha intracellular region, a CD28 intracellular region in tandem 4-1BB intracellular region, a CD28 intracellular region, a 4-1BB intracellular region, a CD28 intracellular region in tandem OX40 intracellular region, a 28-IL2RB-z (YXXQ) intracellular region, a 28-DeltaIL2RB-z (YXXQ) intracellular region, an IL2RB-z (YXXQ) intracellular region, and a DeltaIL2RB-z (YXXQ) intracellular region.
In one or more embodiments, the fusion protein has an amino acid sequence as set forth in any one of SEQ ID NOS 351-606, 1647, 1649, 1651, 1653, 1655, 1657, 1659, 1661, 1663, 1665, 1667, 1671, 1673, and 1675.
In one or more embodiments, the extracellular region of the fusion protein is an extracellular region of TGF- β receptor 3, the transmembrane region being from an IL-7Ralpha transmembrane region or a mutant thereof, and the intracellular domain of the co-stimulatory molecule of the fusion protein is any one selected from the group consisting of an IL-7Ralpha intracellular region, a CD28 intracellular region in tandem 4-1BB intracellular region, a CD28 intracellular region, a 4-1BB intracellular region, a CD28 intracellular region in tandem OX40 intracellular region, a 28-IL2RB-z (YXXQ) intracellular region, a 28-DeltaIL2RB-z (YXXQ) intracellular region, an IL2RB-z (YXXQ) intracellular region, and a DeltaIL2RB-z (YXXQ) intracellular region.
In one or more embodiments, the amino acid sequence of the fusion protein is set forth in any one of SEQ ID NOS 607-862.
The invention also provides a polynucleotide molecule selected from the group consisting of: a polynucleotide molecule or a complementary sequence encoding a fusion protein according to any one of the embodiments of the present invention.
In one or more embodiments, the polynucleotide molecule is selected from any one of SEQ ID NO 863-1630, SEQ ID NO 1648, SEQ ID NO 1650, SEQ ID NO 1652, SEQ ID NO 1654, SEQ ID NO 1656, SEQ ID NO 1658, SEQ ID NO 1660, SEQ ID NO 1662, SEQ ID NO 1664, SEQ ID NO 1666, SEQ ID NO 1668, SEQ ID NO 1670, SEQ ID NO 1672, SEQ ID NO 1674, and SEQ ID NO 1676, or a complement of any one of the polynucleotide molecules shown.
The invention also provides a nucleic acid construct comprising a polynucleotide molecule according to any of the embodiments of the invention.
In one or more embodiments, the nucleic acid construct is a vector.
In one or more embodiments, the vector is an expression vector; preferably a viral vector.
The invention also provides a genetically engineered cell expressing a fusion protein according to any of the embodiments of the invention, and/or carrying a coding sequence for said fusion protein.
In one or more embodiments, the cell is an immune cell.
In one or more embodiments, the immune cell is a T cell.
In one or more embodiments, the cell further expresses a CAR, or carries a coding sequence for a CAR.
In one or more embodiments, the cell further expresses an exogenous TCR, or carries the coding sequence of an exogenous TCR.
The invention also provides the use of the fusion protein, the polynucleotide molecule, the nucleic acid construct and the genetically engineered cell according to any embodiment of the invention in the preparation of a medicament for treating or preventing cancer.
Drawings
FIG. 1: fluorescence micrographs of TCR-T cells electrically transformed with the TCR-EGFP fusion gene and SCR1-5 TCR-T cells co-transformed with the TCR-EGFP fusion gene plus the TGF-beta signal conversion receptor gene.
FIG. 2: in vitro killing effect graph with the effective target ratio of SCR1-5 TCR-T, RIID2 TCR-T and TCR-T to target cell A375 set as 1.5: 1.
FIG. 3: in vitro killing effect graph with the effective target ratio of SCR1-5 TCR-T, RIID TCR-T and TCR-T to target cell A375 set as 1.5: 1.
FIG. 4: in vitro killing effect graph with TCR-T, SCR5TCR-T, RIID TCR-T on target cell A375 at a ratio of 1: 1.
FIG. 5: in vitro killing effect graph with TCR-T, SCR5TCR-T, RIID TCR-T on target cell A375 at ratio of 1: 1.
FIG. 6: TCR-T, SCR6 TCR-T, SCR TCR-T, SCR TCR-T, SCR TCR-T and SCR10 TCR-T versus target cell A375 cell efficacy versus target was set as a plot of in vitro killing activity at 1.5: 1.
FIG. 7: in vitro killing activity plot of TCR-T, SCR10 TCR-T and RIID2 TCR-T versus target cell A375 at 0.5: 1.
FIG. 8: TCR-T, SCR11 TCR-T, SCR TCR-T, SCR TCR-T, SCR TCR-T and SCR15 TCR-T versus target cell A375 cell efficiency target ratio was set as 1.5: 1 in vitro killing activity plot.
FIG. 9: in vitro killing activity plot of TCR-T, SCR15 TCR-T and RIID4 TCR-T versus target cell A375 at 0.75: 1.
FIG. 10: the effect of TCR-T, RIID2 TCR-T, RIID TCR-T and SCR5TCR-T on tumor growth after tumorigenesis in mice of tumor cell line A375.
FIG. 11: the effect of the immune effector cells expressing the TGF-beta signal conversion receptor SCR5, such as TCR-T, on the inhibition of tumor proliferation in vivo is shown.
Detailed Description
It is to be understood that within the scope of the present invention, the above-described technical features of the present invention and the technical features specifically described below (e.g., examples) may be combined with each other to constitute a preferred embodiment.
The invention provides an immune cell for adoptive cell therapy of tumors, which can convert an inhibitory signal for inhibiting the proliferation of the immune cell into a stimulatory signal for enhancing the proliferation of the immune cell after being modified. In some embodiments, the stimulatory signals further comprise a stimulatory signal that enhances anti-tumor activity.
In the present invention, the immune cell has a meaning well known in the art, and refers to a cell involved in or associated with an immune response, including lymphocytes, dendritic cells, monocytes/macrophages, granulocytes, mast cells, and the like. Lymphocytes include T lymphocytes, tumor infiltrating lymphocytes, B lymphocytes, K lymphocytes, and NK lymphocytes. Immune cells suitable for use in the present invention include, inter alia, those typically used in adoptive cell therapy of tumors.
The immune cells of the invention express the signal transduction receptors of the invention, and/or contain coding sequences for the signal transduction receptors. The signal-transducing receptors of the present invention are designed to bind inhibitory molecules but transmit a positive signal rather than an inhibitory signal. That is, these cells convert the "brake" signal into an "acceleration" signal to improve the anti-tumor effect of each immune cell.
Definition of
The present invention uses the following terms. For terms not specifically defined herein, they have meanings well known in the art.
The term "expression cassette" refers to the complete elements required for expression of a gene, including the promoter, gene coding sequence, polyA tailing signal sequence.
The term "coding sequence" is defined herein as that portion of a nucleic acid sequence that directly determines the amino acid sequence of its protein product (e.g., signal-transducing receptor, CAR). The boundaries of the coding sequence are generally determined by a ribosome binding site immediately upstream of the 5 'open reading frame of the mRNA (for prokaryotic cells) and a transcription termination sequence immediately downstream of the 3' open reading frame of the mRNA. A coding sequence can include, but is not limited to, DNA, cDNA, and recombinant nucleic acid sequences.
The term "costimulatory signaling molecule" refers to a molecule present on the surface of an antigen-presenting cell that binds to a costimulatory signaling molecule receptor on a Th cell to produce a costimulatory signal. It can activate the second signal of immune cell, enhance the proliferation capacity of immune cell and the secretion function of cell factor, and prolong the survival time of activated immune cell. The proliferation of lymphocytes requires not only the binding of antigens but also the signaling of co-stimulatory molecules. The transmission of costimulatory signals to T cells is mainly through the expression of the costimulatory molecule CD80 on the surface of antigen presenting cells, with CD86 binding to the CD28 molecule on the surface of T cells. The B cells receive a costimulatory signal via a general pathogen component such as LPS, or via a complement component, or via activated antigen-specific CD40L on the surface of Th cells.
The term "linker" or hinge is a polypeptide fragment that links different proteins or polypeptides, with the purpose of maintaining the linked proteins or polypeptides in their respective spatial conformations, so as to maintain the function or activity of the proteins or polypeptides. Exemplary linkers include G and/or S containing linkers, and, for example, furin 2A peptides.
The term "pharmaceutically acceptable adjuvant" refers to carriers and/or excipients that are pharmacologically and/or physiologically compatible with the subject and active ingredient, which are well known in the art (see, e.g., remington's Pharmaceutical sciences. Edited by geno AR,19th ed. Pennsylvania: pH regulator, surfactant, adjuvant, and ionic strength enhancer. For example, pH adjusting agents include, but are not limited to, phosphate buffers; surfactants include, but are not limited to, cationic, anionic or nonionic surfactants, such as Tween-80; ionic strength enhancers include, but are not limited to, sodium chloride.
The term "effective amount" refers to a dose that achieves treatment, prevention, alleviation and/or amelioration of a disease or disorder described herein in a subject.
The term "disease and/or disorder" refers to a physical condition of the subject that is associated with the disease and/or disorder of the present invention.
The term "subject" can refer to a patient or other animal, particularly a mammal, e.g., a human, dog, monkey, cow, horse, etc., that receives a pharmaceutical composition of the invention to treat, prevent, ameliorate, and/or alleviate a disease or disorder described herein.
The term "tumor stromal cells" refers to some cells in the tumor microenvironment that support the malignant proliferation, anti-apoptosis, invasion, metastasis, escape from immune monitoring and other vital activities of tumor cells, and mainly includes fibroblasts, tumor-associated macrophages (TAMs), regulatory T cells (tregs), undifferentiated bone marrow cells, endothelial cells, pericytes and platelets, endothelial cells and the like.
The term "immunosuppressive cytokine" refers to a molecule produced by tumor cells or tumor stromal cells that acts to suppress immunity, allow tumor cells to escape from immune surveillance of the body, and induce immune tolerance. For example, tumor cell or tumor stromal cell membrane protein PDL1/PDL2 (PD-1 ligand), fasL (Fas ligand), B7-H4 (immune co-stimulatory protein B7-H4, also known as VTCN 1), crry (membrane-associated complement regulatory protein, also known as Cr1 l), HLA-G (non-classical MHC class I molecule), and tumor cell or tumor stromal cell secreted proteins IL13, IL4, TGF-. Beta., IL6, IL8, IL-10, CCL21, IDO (indoleamine 2,3-dioxygenase), VEGF, etc.
The term "extracellular region" refers to a segment of a membrane protein that is located outside of a cell.
The term "domain" refers to a region of a protein having a specific structure and independent function, the number of amino acid residues in a common domain is between 100 and 400, the smallest domain has only 40 to 50 amino acid residues, and the larger domain can exceed 400 amino acid residues.
Signal transduction receptors
The signal-transducing receptor of the present invention is a polypeptide which is a fusion protein comprising the extracellular domain (also referred to as "extracellular region") of a receptor of immunosuppressive cytokines fused to the intracellular domain (also referred to as "intracellular region") of an immunostimulatory molecule (also referred to as "costimulatory signal molecule"). More specifically, the signal transducing receptors of the present invention include the extracellular domain of a receptor for immunosuppressive cytokines, the transmembrane region, and the intracellular domain of a costimulatory signaling molecule. The signal transduction receptors of the present invention bind Th2 or immunosuppressive cytokines but induce immunostimulatory signals rather than immunosuppression, resulting in maintenance of Th1 phenotype, proliferation and cytotoxic characteristics.
In the present invention, the immunosuppressive cytokine may be an immunosuppressive cytokine secreted from a cancer cell or a surrounding tumor stroma, and includes a tumor cell or tumor stroma cell membrane protein and a tumor cell or tumor stroma cell secretory protein. The tumor cell or tumor stroma cell membrane protein includes but is not limited to PDL1/PDL2 (PD-1 ligand), fasL (Fas ligand), B7-H4 (immune co-stimulatory protein B7-H4, also known as VTCN 1), crry (membrane associated complement regulatory protein, also known as Cr1 l) and HLA-G (non-classical MHC class I molecule); the tumor cell or tumor stromal cell secreted protein includes but is not limited to IL13, IL4, TGF-beta, IL6, IL8, IL-10, CCL21, IDO (indoleamine 2,3-dioxygenase), VEGF and the like. The present invention relates to the construction of signal transduction receptors of the invention using the extracellular domains of these immunosuppressive cytokine receptors or functional fragments thereof or mutants that retain the ability to bind to the corresponding immunosuppressive cytokines.
In a particularly preferred embodiment, the receptor for an immunosuppressive cytokine in the signaling receptor of the invention is the TGF- β receptor. The extracellular domain of a TGF- β receptor of the present invention may comprise only the extracellular domain of a TGF- β receptor, or may be a chimeric TGF- β receptor comprising a TGF-binding fragment of a TGF- β receptor. The extracellular domains of any isoform of a native TGF-beta receptor may be used, including TGF-beta receptor 1, TGF-beta receptor 2, and TGF-beta receptor 3, with TGF-beta receptor 1 and TGF-beta receptor 2 being preferred. The amino acid sequence and polynucleotide sequence of the extracellular region of exemplary TGF-beta receptor 1 can be shown in SEQ ID NO 1 and 2, respectively. Exemplary nucleotide sequences encoding TGF-beta receptor 2 include sequences shown in Genbank accession nos. NM _001024847.2 or NM _003242.5, and amino acid sequences thereof are Genbank accession nos. NP _001020018 or NM _003242, respectively. In some embodiments, the extracellular domain of the invention comprises amino acids 23-166 from TGF-beta receptor 2, as shown in SEQ ID NO 3, and the coding sequence is shown in SEQ ID NO 4. The amino acid sequence and polynucleotide sequence of the extracellular region of exemplary TGF-beta receptor 3 may be shown in SEQ ID NOS: 5 and 6, respectively.
In other embodiments, the receptor for an immunosuppressive factor in a signal-transducing receptor of the invention is an IL13 receptor, and the extracellular domain is an extracellular domain of an IL13 receptor, or a chimeric IL13 receptor comprising an IL 13-binding fragment of an IL13 receptor.
In the present invention, co-stimulatory signaling molecules include CD28, CD134 (OX 40), CD137 (4-1 BB), LCK, ICOS, DAP10, siglec-9, siglec-10, siglec-15, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, IL-2R, IL-4R, IL-7R, IL-10R, IL-12R, IL-15R, IL-21R, CD and CD40. The signal transduction receptors of the present invention may be constructed using the intracellular domain (intracellular domain) of one or more of these costimulatory signaling molecules or functional fragments thereof or mutants that retain the biological function of the costimulatory signaling molecules to deliver costimulatory signals to activate immune cells. An exemplary IL-7R can be IL-7Ralpha, an exemplary amino acid sequence and corresponding coding sequence of which can be set forth in SEQ ID NOS: 7 and 8, respectively. The amino acid sequences of the exemplary intracellular region of CD28 and the corresponding coding sequence can be shown in SEQ ID NOS: 9 and 10, respectively. The amino acid sequences of the exemplary intracellular region of 4-1BB and the corresponding coding sequences can be shown in SEQ ID NOS: 11 and 12, respectively. The amino acid sequence of OX40 intracellular domain is shown as SEQ ID NO:1645, and the coding sequence is shown as SEQ ID NO: 1646.
In the present invention, the extracellular domain of the receptor for immunosuppressive cytokines is linked to the intracellular domain of the costimulatory signaling molecule through a transmembrane region (transmembrane domain). The transmembrane region may be of any origin. Transmembrane regions suitable for use in the present invention include, but are not limited to, any one or more of CD28, CD8, CD134 (OX 40), CD137 (4-1 BB), LCK, ICOS, DAP10, siglec-9, siglec-10, siglec-15, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, IL-2Rgamma, IL-2Rbeta, IL-4Ralpha, IL-7Ralpha, IL-10R, IL-12R, IL-15R, IL-21R, CD and a transmembrane region of CD40 or a mutant thereof that retains transmembrane function. The transmembrane region may be derived from the same molecule as the intracellular domain or from a different molecule. In the present invention, the preferred transmembrane region is derived from the IL-7Ralpha transmembrane region and mutants thereof. Exemplary amino acid sequences and coding sequences of the IL-7Ralpha transmembrane region are shown in SEQ ID NOS: 27 and 28, respectively. Exemplary amino acid sequences of mutants of the IL-7Ralpha transmembrane region may be selected from any one of SEQ ID NOs 29-60; preferably, the coding nucleic acid sequence is selected from any one of SEQ ID NO 61-94.
It is understood that "functional fragment" as used herein refers to a fragment that retains a desired biological function. For example, a functional fragment of an extracellular domain as described herein refers to a functional fragment that retains the ability to bind the corresponding immunosuppressive cytokine; a functional fragment of an intracellular domain refers to a fragment that retains the biological function of the costimulatory signaling molecule to deliver a costimulatory signal to activate an immune cell. For example, a functional fragment of the extracellular domain of TGF- β receptor 2 is a functional fragment from the extracellular domain that binds TGF- β receptor 2. Functional fragments of each extracellular domain as well as functional fragments of each intracellular domain suitable for use in the present invention can be readily determined by one skilled in the art in combination with prior art means of the art.
"mutant" as used herein includes mutants of each of the extracellular, transmembrane and intracellular domains, so long as the mutant retains the respective biological functions of the extracellular, transmembrane and intracellular domains. For example, suitable mutants of an extracellular domain for use in the present invention include mutants having at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to the extracellular domain in comparison; mutants of a transmembrane region suitable for use in the invention include mutants having at least 80%, at least 85%, at least 90%, at least 95%, at least 97% or at least 99% sequence identity to the transmembrane region as compared; mutants of an intracellular domain suitable for use in the present invention include mutants having at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to the intracellular domain in comparison. Alternatively, the mutants of the invention have one or more (e.g., within 20, within 15, within 10, within 8, within 5, or within 3, e.g., 1-20, 1-10, etc.) amino acid residue insertions, substitutions, or deletions as compared to the compared sequence.
The invention also includes mutants of the signal transduction receptors described hereinbefore, such as mutants having at least 80%, at least 85%, at least 90%, at least 95%, at least 97% or at least 99% sequence identity to the signal transduction receptor. More specifically, the present invention includes mutants having one or more (e.g., within 20, within 15, within 10, within 8, within 5, or within 3, such as 1-20, 1-10, etc.) amino acid residue insertions, substitutions, or deletions compared to the signal transducing receptors described above. Such mutants retain the biological functions of the signal transduction receptors of the present invention, including but not limited to, converting inhibitory signals that inhibit immune cell proliferation into stimulatory signals that enhance immune cell proliferation. Mutations can occur in any one, any two, or all three of the extracellular, transmembrane, and intracellular domains described herein.
Exemplary signal transducing receptors of the present invention include, but are not limited to, signal transducing receptors comprising or consisting of the extracellular, transmembrane and intracellular domains shown in each row of table 1 below:
TABLE 1
Extracellular domain Transmembrane region Intracellular domains
TGF-beta receptor 1 CD28 OX40
TGF-beta receptor 1 CD28 4-1BB
TGF-beta receptor 1 CD28 CD8
TGF-beta receptor 1 CD28 OX40+4-1BB
TGF-beta receptor 1 CD28 CD8+4-1BB
TGF-beta receptor 1 CD28 CD8+OX40
TGF-beta receptor 1 CD28 OX40+CD3z
TGF-beta receptor 1 CD28 4-1BB+CD3z
TGF-beta receptor 1 CD28 CD8+CD3z
TGF-beta receptor 1 CD28 OX40+4-1BB+CD3z
TGF-beta receptor 1 CD28 CD8+4-1BB+CD3z
TGF-beta receptor 1 CD28 CD8+OX40+CD3z
TGF-beta receptor 1 CD8 OX40
TGF-beta receptor 1 CD8 4-1BB
TGF-beta receptor 1 CD8 CD28
TGF-beta receptor 1 CD8 OX40+4-1BB
TGF-beta receptor 1 CD8 CD28+4-1BB
TGF-beta receptor 1 CD8 CD28+OX40
TGF-beta receptor 1 CD8 OX40+CD3z
TGF-beta receptor 1 CD8 4-1BB+CD3z
TGF-beta receptor 1 CD8 CD28+CD3z
TGF-beta receptor 1 CD8 OX40+4-1BB+CD3z
TGF-beta receptor 1 CD8 CD28+4-1BB+CD3z
TGF-beta receptor 1 CD8 CD28+OX40+CD3z
TGF-beta receptor 1 4-1BB CD28
TGF-beta receptor 1 4-1BB OX40
TGF-beta receptor 1 4-1BB CD28+OX40
TGF-beta receptor 1 IL-2Rgamma 4-1BB
TGF-beta receptor 1 IL-2Rgamma CD28
TGF-beta receptor 1 IL-2Rgamma OX40
TGF-beta receptor 1 IL-2Rgamma CD28+4-1BB
TGF-beta receptor 1 IL-2Rgamma CD28+4-1BB+CD3z
TGF-beta receptor 1 IL-2Rgamma IL-2Rgamma
TGF-beta receptor 1 IL-2Rbeta 4-1BB
TGF-beta receptor 1 IL-2Rbeta CD28
TGF-beta receptor 1 IL-2Rbeta OX40
TGF-beta receptor 1 IL-2Rbeta CD28+4-1BB
TGF-beta receptor 1 IL-2Rbeta CD28+4-1BB+CD3z
TGF-beta receptor 1 IL-2Rbeta IL-2Rbeta
TGF-beta receptor 1 IL-4Ralpha CD28
TGF-beta receptor 1 IL-4Ralpha 4-1BB
TGF-beta receptor 1 IL-4Ralpha OX40
TGF-beta receptor 1 IL-4Ralpha CD28+4-1BB
TGF-beta receptor 1 IL-4Ralpha CD28+4-1BB+CD3z
TGF-beta receptor 1 IL-4Ralpha IL-4Ralpha
TGF-beta receptor 1 IL-7Ralpha CD28
TGF-beta receptor 1 IL-7Ralpha 4-1BB
TGF-beta receptor 1 IL-7Ralpha OX40
TGF-beta receptor 1 IL-7Ralpha CD28+4-1BB
TGF-beta receptor 1 IL-7Ralpha CD28+4-1BB+CD3z
TGF-beta receptor 1 IL-7Ralpha IL-7Ralpha
TGF-beta receptor 1 IL-10R CD28
TGF-beta receptor 1 IL-10R 4-1BB
TGF-beta receptor 1 IL-10R OX40
TGF-beta receptor 1 IL-10R CD28+4-1BB
TGF-beta receptor 1 IL-10R CD28+4-1BB+CD3z
TGF-beta receptor 1 IL-10R IL-10R
TGF-beta receptor 1 IL-12R CD28
TGF-beta receptor 1 IL-12R 4-1BB
TGF-beta receptor 1 IL-12R OX40
TGF-beta receptor 1 IL-12R CD28+4-1BB
TGF-beta receptor 1 IL-12R CD28+4-1BB+CD3z
TGF-beta receptor 1 IL-12R IL-12R
TGF-beta receptor 1 IL-15R CD28
TGF-beta receptor 1 IL-15R 4-1BB
TGF-beta receptor 1 IL-15R OX40
TGF-beta receptor 1 IL-15R CD28+4-1BB
TGF-beta receptor 1 IL-15R CD28+4-1BB+CD3z
TGF-beta receptor 1 IL-15R IL-15R
TGF-beta receptor 1 IL-21R CD28
TGF-beta receptor 1 IL-21R 4-1BB
TGF-beta receptor 1 IL-21R OX40
TGF-beta receptor 1 IL-21R CD28+4-1BB
TGF-beta receptor 1 IL-21R CD28+4-1BB+CD3z
TGF-beta receptor 1 IL-21R IL-21R
TGF-beta receptor 1 CD40 CD28
TGF-beta receptor 1 CD40 4-1BB
TGF-beta receptor 1 CD40 OX40
TGF-beta receptor 1 CD40 CD28+4-1BB
TGF-beta receptor 1 CD40 CD28+4-1BB+CD3z
TGF-beta receptor 1 CD40 CD40
TGF-beta receptor 1 DAP10 CD28
TGF-beta receptor 1 DAP10 4-1BB
TGF-beta receptor 1 DAP10 OX40
TGF-beta receptor 1 DAP10 CD28+4-1BB
TGF-beta receptor 1 DAP10 CD28+4-1BB+CD3z
TGF-beta receptor 1 DAP10 DAP10
TGF-beta receptor 1 ICOS CD28
TGF-beta receptor 1 ICOS 4-1BB
TGF-beta receptor 1 ICOS OX40
TGF-beta receptor 1 ICOS CD28+4-1BB
TGF-beta receptor 1 ICOS CD28+4-1BB+CD3z
TGF-beta receptor 1 ICOS ICOS
TGF-beta receptor 2 CD28 OX40
TGF-beta receptor 2 CD28 4-1BB
TGF-beta receptor 2 CD28 CD8
TGF-beta receptor 2 CD28 OX40+4-1BB
TGF-beta receptor 2 CD28 CD8+4-1BB
TGF-beta receptor 2 CD28 CD8+OX40
TGF-beta receptor 2 CD28 OX40+CD3z
TGF-beta receptor 2 CD28 4-1BB+CD3z
TGF-beta receptor 2 CD28 CD8+CD3z
TGF-beta receptor 2 CD28 OX40+4-1BB+CD3z
TGF-beta receptor 2 CD28 CD8+4-1BB+CD3z
TGF-beta receptor 2 CD28 CD8+OX40+CD3z
TGF-beta receptor 2 CD8 OX40
TGF-beta receptor 2 CD8 4-1BB
TGF-beta receptor 2 CD8 CD28
TGF-beta receptor 2 CD8 OX40+4-1BB
TGF-beta receptor 2 CD8 CD28+4-1BB
TGF-beta receptor 2 CD8 CD28+OX40
TGF-beta receptor 2 CD8 OX40+CD3z
TGF-beta receptor 2 CD8 4-1BB+CD3z
TGF-beta receptor 2 CD8 CD28+CD3z
TGF-beta receptor 2 CD8 OX40+4-1BB+CD3z
TGF-beta receptor 2 CD8 CD28+4-1BB+CD3z
TGF-beta receptor 2 CD8 CD28+OX40+CD3z
TGF-beta receptor 2 4-1BB CD28
TGF-beta receptor 2 4-1BB OX40
TGF-beta receptor 2 4-1BB CD28+OX40
TGF-beta receptor 2 IL-2Rgamma 4-1BB
TGF-beta receptor 2 IL-2Rgamma CD28
TGF-beta receptor 2 IL-2Rgamma OX40
TGF-beta receptor 2 IL-2Rgamma CD28+4-1BB
TGF-beta receptor 2 IL-2Rgamma CD28+4-1BB+CD3z
TGF-beta receptor 2 IL-2Rgamma IL-2Rgamma
TGF-beta receptor 2 IL-2Rbeta 4-1BB
TGF-beta receptor 2 IL-2Rbeta CD28
TGF-beta receptor 2 IL-2Rbeta OX40
TGF-beta receptor 2 IL-2Rbeta CD28+4-1BB
TGF-beta receptor 2 IL-2Rbeta CD28+4-1BB+CD3z
TGF-beta receptor 2 IL-2Rbeta IL-2Rbeta
TGF-beta receptor 2 IL-4Ralpha CD28
TGF-beta receptor 2 IL-4Ralpha 4-1BB
TGF-beta receptor 2 IL-4Ralpha OX40
TGF-beta receptor 2 IL-4Ralpha CD28+4-1BB
TGF-beta receptor 2 IL-4Ralpha CD28+4-1BB+CD3z
TGF-beta receptor 2 IL-4Ralpha IL-4Ralpha
TGF-beta receptor 2 IL-7Ralpha CD28
TGF-beta receptor 2 IL-7Ralpha 4-1BB
TGF-beta receptor 2 IL-7Ralpha OX40
TGF-beta receptor 2 IL-7Ralpha CD28+4-1BB
TGF-beta receptor 2 IL-7Ralpha CD28+4-1BB+CD3z
TGF-beta receptor 2 IL-7Ralpha IL-7Ralpha
TGF-beta receptor 2 IL-10R CD28
TGF-beta receptor 2 IL-10R 4-1BB
TGF-beta receptor 2 IL-10R OX40
TGF-beta receptor 2 IL-10R CD28+4-1BB
TGF-beta receptor 2 IL-10R CD28+4-1BB+CD3z
TGF-beta receptor 2 IL-10R IL-10R
TGF-beta receptor 2 IL-12R CD28
TGF-beta receptor 2 IL-12R 4-1BB
TGF-beta receptor 2 IL-12R OX40
TGF-beta receptor 2 IL-12R CD28+4-1BB
TGF-beta receptor 2 IL-12R CD28+4-1BB+CD3z
TGF-beta receptor 2 IL-12R IL-12R
TGF-beta receptor 2 IL-15R CD28
TGF-beta receptor 2 IL-15R 4-1BB
TGF-beta receptor 2 IL-15R OX40
TGF-beta receptor 2 IL-15R CD28+4-1BB
TGF-beta receptor 2 IL-15R CD28+4-1BB+CD3z
TGF-beta receptor 2 IL-15R IL-15R
TGF-beta receptor 2 IL-21R CD28
TGF-beta receptor 2 IL-21R 4-1BB
TGF-beta receptor 2 IL-21R OX40
TGF-beta receptor 2 IL-21R CD28+4-1BB
TGF-beta receptor 2 IL-21R CD28+4-1BB+CD3z
TGF-beta receptor 2 IL-21R IL-21R
TGF-beta receptor 2 CD40 CD28
TGF-beta receptor 2 CD40 4-1BB
TGF-beta receptor 2 CD40 OX40
TGF-beta receptor 2 CD40 CD28+4-1BB
TGF-beta receptor 2 CD40 CD28+4-1BB+CD3z
TGF-beta receptor 2 CD40 CD40
TGF-beta receptor 2 DAP10 CD28
TGF-beta receptor 2 DAP10 4-1BB
TGF-beta receptor 2 DAP10 OX40
TGF-beta receptor 2 DAP10 CD28+4-1BB
TGF-beta receptor 2 DAP10 CD28+4-1BB+CD3z
TGF-beta receptor 2 DAP10 DAP10
TGF-beta receptor 2 ICOS CD28
TGF-beta receptor 2 ICOS 4-1BB
TGF-beta receptor 2 ICOS OX40
TGF-beta receptor 2 ICOS CD28+4-1BB
TGF-beta receptor 2 ICOS CD28+4-1BB+CD3z
TGF-beta receptor 2 ICOS ICOS
TGF-beta receptor 3 CD28 OX40
TGF-beta receptor 3 CD28 4-1BB
TGF-beta receptor 3 CD28 CD8
TGF-beta receptor 3 CD28 OX40+4-1BB
TGF-beta receptor 3 CD28 CD8+4-1BB
TGF-beta receptor 3 CD28 CD8+OX40
TGF-beta receptor 3 CD28 OX40+CD3z
TGF-beta receptor 3 CD28 4-1BB+CD3z
TGF-beta receptor 3 CD28 CD8+CD3z
TGF-beta receptor 3 CD28 OX40+4-1BB+CD3z
TGF-beta receptor 3 CD28 CD8+4-1BB+CD3z
TGF-beta receptor 3 CD28 CD8+OX40+CD3z
TGF-beta receptor 3 CD8 OX40
TGF-beta receptor 3 CD8 4-1BB
TGF-beta receptor 3 CD8 CD28
TGF-beta receptor 3 CD8 OX40+4-1BB
TGF-beta receptor 3 CD8 CD28+4-1BB
TGF-beta receptor 3 CD8 CD28+OX40
TGF-beta receptor 3 CD8 OX40+CD3z
TGF-beta receptor 3 CD8 4-1BB+CD3z
TGF-beta receptor 3 CD8 CD28+CD3z
TGF-beta receptor 3 CD8 OX40+4-1BB+CD3z
TGF-beta receptor 3 CD8 CD28+4-1BB+CD3z
TGF-beta receptor 3 CD8 CD28+OX40+CD3z
TGF-beta receptor 3 4-1BB CD28
TGF-beta receptor 3 4-1BB OX40
TGF-beta receptor 3 4-1BB CD28+OX40
TGF-beta receptor 3 IL-2Rgamma 4-1BB
TGF-beta receptor 3 IL-2Rgamma CD28
TGF-beta receptor 3 IL-2Rgamma OX40
TGF-beta receptor 3 IL-2Rgamma CD28+4-1BB
TGF-beta receptor 3 IL-2Rgamma CD28+4-1BB+CD3z
TGF-beta receptor 3 IL-2Rgamma IL-2Rgamma
TGF-beta receptor 3 IL-2Rbeta 4-1BB
TGF-beta receptor 3 IL-2Rbeta CD28
TGF-beta receptor 3 IL-2Rbeta OX40
TGF-beta receptor 3 IL-2Rbeta CD28+4-1BB
TGF-beta receptor 3 IL-2Rbeta CD28+4-1BB+CD3z
TGF-beta receptor 3 IL-2Rbeta IL-2Rbeta
TGF-beta receptor 3 IL-4Ralpha CD28
TGF-beta receptor 3 IL-4Ralpha 4-1BB
TGF-beta receptor 3 IL-4Ralpha OX40
TGF-beta receptor 3 IL-4Ralpha CD28+4-1BB
TGF-beta receptor 3 IL-4Ralpha CD28+4-1BB+CD3z
TGF-beta receptor 3 IL-4Ralpha IL-4Ralpha
TGF-beta receptor 3 IL-7Ralpha CD28
TGF-beta receptor 3 IL-7Ralpha 4-1BB
TGF-beta receptor 3 IL-7Ralpha OX40
TGF-beta receptor 3 IL-7Ralpha CD28+4-1BB
TGF-beta receptor 3 IL-7Ralpha CD28+4-1BB+CD3z
TGF-beta receptor 3 IL-7Ralpha IL-7Ralpha
TGF-beta receptor 3 IL-10R CD28
TGF-beta receptor 3 IL-10R 4-1BB
TGF-beta receptor 3 IL-10R OX40
TGF-beta receptor 3 IL-10R CD28+4-1BB
TGF-beta receptor 3 IL-10R CD28+4-1BB+CD3z
TGF-beta receptor 3 IL-10R IL-10R
TGF-beta receptor 3 IL-12R CD28
TGF-beta receptor 3 IL-12R 4-1BB
TGF-beta receptor 3 IL-12R OX40
TGF-beta receptor 3 IL-12R CD28+4-1BB
TGF-beta receptor 3 IL-12R CD28+4-1BB+CD3z
TGF-beta receptor 3 IL-12R IL-12R
TGF-beta receptor 3 IL-15R CD28
TGF-beta receptor 3 IL-15R 4-1BB
TGF-beta receptor 3 IL-15R OX40
TGF-beta receptor 3 IL-15R CD28+4-1BB
TGF-beta receptor 3 IL-15R CD28+4-1BB+CD3z
TGF-beta receptor 3 IL-15R IL-15R
TGF-beta receptor 3 IL-21R CD28
TGF-beta receptor 3 IL-21R 4-1BB
TGF-beta receptor 3 IL-21R OX40
TGF-beta receptor 3 IL-21R CD28+4-1BB
TGF-beta receptor 3 IL-21R CD28+4-1BB+CD3z
TGF-beta receptor 3 IL-21R IL-21R
TGF-beta receptor 3 CD40 CD28
TGF-beta receptor 3 CD40 4-1BB
TGF-beta receptor 3 CD40 OX40
TGF-beta receptor 3 CD40 CD28+4-1BB
TGF-beta receptor 3 CD40 CD28+4-1BB+CD3z
TGF-beta receptor 3 CD40 CD40
TGF-beta receptor 3 DAP10 CD28
TGF-beta receptor 3 DAP10 4-1BB
TGF-beta receptor 3 DAP10 OX40
TGF-beta receptor 3 DAP10 CD28+4-1BB
TGF-beta receptor 3 DAP10 CD28+4-1BB+CD3z
TGF-beta receptor 3 DAP10 DAP10
TGF-beta receptor 3 ICOS CD28
TGF-beta receptor 3 ICOS 4-1BB
TGF-beta receptor 3 ICOS OX40
TGF-beta receptor 3 ICOS CD28+4-1BB
TGF-beta receptor 3 ICOS CD28+4-1BB+CD3z
TGF-beta receptor 3 ICOS ICOS
In the present invention, it may also comprise a mutated activated receptor transmembrane region sequence, preferably comprising a mutant of the mutated IL-7Ralpha transmembrane region. The mutant of the transmembrane region of the activated receptor can better lead the activated receptor to be aggregated, and further activate and amplify the intracellular signal on the basis of the wild type receptor. The sequences of IL-7Ralpha transmembrane domain mutant 1-32 (IL-7 Ralpha TM mut 1-32) are shown in tables 2 and 3 below:
table 2: IL-7Ralpha transmembrane region mutant 1-32 amino acid sequence
Name(s) Sequence of Serial number
IL-7Ralpha TM mut1 PILLPPCLTISILSFFSVALLVILACVLW SEQ ID NO:29
IL-7Ralpha TM mut2 PILLTISKCHILSFFSVALLVILACVLW SEQ ID NO:30
IL-7Ralpha TM mut3 PIFSCGPLLTISILSFFSVALLVILACVLW SEQ ID NO:31
IL-7Ralpha TM mut4 PILLTCHLISILSFFSVALLVILACVLW SEQ ID NO:32
IL-7Ralpha TM mut5 PILLTPPVCSVTISILSFFSVALLVILACVLW SEQ ID NO:33
IL-7Ralpha TM mut6 PILLFCRKDTISILSFFSVALLVILACVLW SEQ ID NO:34
IL-7Ralpha TM mut7 PILLRCTISILSFFSVALLVILACVLW SEQ ID NO:35
IL-7Ralpha TM mut8 PILPCPLTISILSFFSVALLVILACVLW SEQ ID NO:36
IL-7Ralpha TM mut9 PILLMPEQDCPTISILSFFSVALLVILACVLW SEQ ID NO:37
IL-7Ralpha TM mut10 PILCPPLTISILSFFSVALLVILACVLW SEQ ID NO:38
IL-7Ralpha TM mut11 PILRCPSTISILSFFSVALLVILACVLW SEQ ID NO:39
IL-7Ralpha TM mut12 PIFDCIGVLTISILSFFSVALLVILACVLW SEQ ID NO:40
IL-7Ralpha TM mut13 PILLTHRGCILSFFSVALLVILACVLW SEQ ID NO:41
IL-7Ralpha TM mut14 PILPLCSAISILSFFSVALLVILACVLW SEQ ID NO:42
IL-7Ralpha TM mut15 PILLTISILSFFSVGEAALLVILACVLW SEQ ID NO:43
IL-7Ralpha TM mut16 PILPIYRCVLISILSFFSVALLVILACVLW SEQ ID NO:44
IL-7Ralpha TM mut17 PIFECLTISILSFFSVALLVILACVLW SEQ ID NO:45
IL-7Ralpha TM mut18 PIFTCPSISILSFFSVALLVILACVLW SEQ ID NO:46
IL-7Ralpha TM mut19 PILCPSPTISILSFFSVALLVILACVLW SEQ ID NO:47
IL-7Ralpha TM mut20 PILLTISILSFFSVEKVLVILACVLW SEQ ID NO:48
IL-7Ralpha TM mut21 PILLTSHQPCILSFFSVALLVILACVLW SEQ ID NO:49
IL-7Ralpha TM mut22 PILLTISILSCSTISILSFFSVALLVILACVLW SEQ ID NO:50
IL-7Ralpha TM mut23 PILLTISICQSVALLVILACVLW SEQ ID NO:51
IL-7Ralpha TM mut24 PICGIREIISILSFFSVALLVILACVLW SEQ ID NO:52
IL-7Ralpha TM mut25 PILLTISILSFFSGALLVILACVLW SEQ ID NO:53
IL-7Ralpha TM mut26 PILLGCTISILSFFSVALLVILACVLW SEQ ID NO:54
IL-7Ralpha TM mut27 PCRPHLTISILSFFSVALLVILACVLW SEQ ID NO:55
IL-7Ralpha TM mut28 PILGCITISILSFFSVALLVILACVLW SEQ ID NO:56
IL-7Ralpha TM mut29 PILLTRPCGISILSFFSVALLVILACVLW SEQ ID NO:57
IL-7Ralpha TM mut30 PILLTLPCVTISILSFFSVALLVILACVLW SEQ ID NO:58
IL-7Ralpha TM mut31 PICWMKLTISILSFFSVALLVILACVLW SEQ ID NO:59
IL-7Ralpha TM mut32 PILLKKCTNISILSFFSVALLVILACVLW SEQ ID NO:60
Table 3: IL-7Ralpha transmembrane region mutant 1-32 nucleotide sequence
Figure PCTCN2021097361-APPB-000001
Figure PCTCN2021097361-APPB-000002
Figure PCTCN2021097361-APPB-000003
Exemplary signaling receptors of the present invention comprising IL-7Ralpha transmembrane domain mutants 1-32 include, but are not limited to, the signaling receptors shown in any of the rows in Table 4 below:
TABLE 4
Extracellular region Transmembrane region Intracellular region Serial number (protein) Sequence No. (nucleic acid)
TGF-beta receptor 1 IL-7Ralpha TM mut1 Intracellular domain of IL-7Ralpha SEQ ID NO:95 SEQ ID NO:863
TGF-beta receptor 1 IL-7Ralpha TM mut2 Intracellular domain of IL-7Ralpha SEQ ID NO:96 SEQ ID NO:864
TGF-beta receptor 1 IL-7Ralpha TM mut3 Intracellular domain of IL-7Ralpha SEQ ID NO:97 SEQ ID NO:865
TGF-beta receptor 1 IL-7Ralpha TM mut4 Intracellular domain of IL-7Ralpha SEQ ID NO:98 SEQ ID NO:866
TGF-beta receptor 1 IL-7Ralpha TM mut5 Intracellular domain of IL-7Ralpha SEQ ID NO:99 SEQ ID NO:867
TGF-beta receptor 1 IL-7Ralpha TM mut6 Intracellular domain of IL-7Ralpha SEQ ID NO:100 SEQ ID NO:868
TGF-beta receptor 1 IL-7Ralpha TM mut7 Intracellular domain of IL-7Ralpha SEQ ID NO:101 SEQ ID NO:869
TGF-beta receptor 1 IL-7Ralpha TM mut8 Intracellular domain of IL-7Ralpha SEQ ID NO:102 SEQ ID NO:870
TGF-beta receptor 1 IL-7Ralpha TM mut9 Intracellular domain of IL-7Ralpha SEQ ID NO:103 SEQ ID NO:871
TGF-beta receptor 1 IL-7Ralpha TM mut10 Intracellular domain of IL-7Ralpha SEQ ID NO:104 SEQ ID NO:872
TGF-beta receptor 1 IL-7Ralpha TM mut11 Intracellular domain of IL-7Ralpha SEQ ID NO:105 SEQ ID NO:873
TGF-beta receptor 1 IL-7Ralpha TM mut12 Intracellular domain of IL-7Ralpha SEQ ID NO:106 SEQ ID NO:874
TGF-beta receptor 1 IL-7Ralpha TM mut13 Intracellular domain of IL-7Ralpha SEQ ID NO:107 SEQ ID NO:875
TGF-beta receptor 1 IL-7Ralpha TM mut14 Intracellular domain of IL-7Ralpha SEQ ID NO:108 SEQ ID NO:876
TGF-beta receptor 1 IL-7Ralpha TM mut15 Intracellular domain of IL-7Ralpha SEQ ID NO:109 SEQ ID NO:877
TGF-beta receptor 1 IL-7Ralpha TM mut16 Intracellular domain of IL-7Ralpha SEQ ID NO:110 SEQ ID NO:878
TGF-beta receptor 1 IL-7Ralpha TM mut17 Intracellular domain of IL-7Ralpha SEQ ID NO:111 SEQ ID NO:879
TGF-beta receptor 1 IL-7Ralpha TM mut18 Intracellular domain of IL-7Ralpha SEQ ID NO:112 SEQ ID NO:880
TGF-beta receptor 1 IL-7Ralpha TM mut19 Intracellular domain of IL-7Ralpha SEQ ID NO:113 SEQ ID NO:881
TGF-beta receptor 1 IL-7Ralpha TM mut20 Intracellular domain of IL-7Ralpha SEQ ID NO:114 SEQ ID NO:882
TGF-beta receptor 1 IL-7Ralpha TM mut21 Intracellular domain of IL-7Ralpha SEQ ID NO:115 SEQ ID NO:883
TGF-beta receptor 1 IL-7Ralpha TM mut22 Intracellular domain of IL-7Ralpha SEQ ID NO:116 SEQ ID NO:884
TGF-beta receptor 1 IL-7Ralpha TM mut23 Intracellular domain of IL-7Ralpha SEQ ID NO:117 SEQ ID NO:885
TGF-beta receptor 1 IL-7Ralpha TM mut24 Intracellular domain of IL-7Ralpha SEQ ID NO:118 SEQ ID NO:886
TGF-beta receptor 1 IL-7Ralpha TM mut25 Intracellular domain of IL-7Ralpha SEQ ID NO:119 SEQ ID NO:887
TGF-beta receptor 1 IL-7Ralpha TM mut26 Intracellular domain of IL-7Ralpha SEQ ID NO:120 SEQ ID NO:888
TGF-beta receptor 1 IL-7Ralpha TM mut27 Intracellular domain of IL-7Ralpha SEQ ID NO:121 SEQ ID NO:889
TGF-beta receptor 1 IL-7Ralpha TM mut28 Intracellular domain of IL-7Ralpha SEQ ID NO:122 SEQ ID NO:890
TGF-beta receptor 1 IL-7Ralpha TM mut29 Intracellular domain of IL-7Ralpha SEQ ID NO:123 SEQ ID NO:891
TGF-beta receptor 1 IL-7Ralpha TM mut30 Intracellular domain of IL-7Ralpha SEQ ID NO:124 SEQ ID NO:892
TGF-beta receptor 1 IL-7Ralpha TM mut31 Intracellular domain of IL-7Ralpha SEQ ID NO:125 SEQ ID NO:893
TGF-beta receptor 1 IL-7Ralpha TM mut32 Intracellular domain of IL-7Ralpha SEQ ID NO:126 SEQ ID NO:894
TGF-beta receptor 1 IL-7Ralpha TM mut1 CD28 intracellular +4-1BB intracellular SEQ ID NO:127 SEQ ID NO:895
TGF-beta receptor 1 IL-7Ralpha TM mut2 CD28 intracellular +4-1BB intracellular SEQ ID NO:128 SEQ ID NO:896
TGF-beta receptor 1 IL-7Ralpha TM mut3 CD28 intracellular +4-1BB intracellular SEQ ID NO:129 SEQ ID NO:897
TGF-beta receptor 1 IL-7Ralpha TM mut4 CD28 intracellular +4-1BB intracellular SEQ ID NO:130 SEQ ID NO:898
TGF-beta receptor 1 IL-7Ralpha TM mut5 CD28 intracellular +4-1BB intracellular SEQ ID NO:131 SEQ ID NO:899
TGF-beta receptor 1 IL-7Ralpha TM mut6 CD28 intracellular +4-1BB intracellular SEQ ID NO:132 SEQ ID NO:900
TGF-beta receptor 1 IL-7Ralpha TM mut7 CD28 intracellular +4-1BB intracellular SEQ ID NO:133 SEQ ID NO:901
TGF-beta receptor 1 IL-7Ralpha TM mut8 CD28 intracellular +4-1BB intracellular SEQ ID NO:134 SEQ ID NO:902
TGF-beta receptor 1 IL-7Ralpha TM mut9 CD28 intracellular +4-1BB intracellular SEQ ID NO:135 SEQ ID NO:903
TGF-beta receptor 1 IL-7Ralpha TM mut10 CD28 intracellular +4-1BB intracellular SEQ ID NO:136 SEQ ID NO:904
TGF-beta receptor 1 IL-7Ralpha TM mut11 CD28 intracellular +4-1BB intracellular SEQ ID NO:137 SEQ ID NO:905
TGF-beta receptor 1 IL-7Ralpha TM mut12 CD28 intracellular +4-1BB intracellular SEQ ID NO:138 SEQ ID NO:906
TGF-beta receptor 1 IL-7Ralpha TM mut13 CD28 intracellular +4-1BB intracellular SEQ ID NO:139 SEQ ID NO:907
TGF-beta receptor 1 IL-7Ralpha TM mut14 CD28 intracellular +4-1BB intracellular SEQ ID NO:140 SEQ ID NO:908
TGF-beta receptor 1 IL-7Ralpha TM mut15 CD28 intracellular +4-1BB intracellular SEQ ID NO:141 SEQ ID NO:909
TGF-beta receptor 1 IL-7Ralpha TM mut16 CD28 intracellular +4-1BB intracellular SEQ ID NO:142 SEQ ID NO:910
TGF-beta receptor 1 IL-7Ralpha TM mut17 CD28 intracellular +4-1BB intracellular SEQ ID NO:143 SEQ ID NO:911
TGF-beta receptor 1 IL-7Ralpha TM mut18 CD28 intracellular +4-1BB intracellular SEQ ID NO:144 SEQ ID NO:912
TGF-beta receptor 1 IL-7Ralpha TM mut19 CD28 intracellular +4-1BB intracellular SEQ ID NO:145 SEQ ID NO:913
TGF-beta receptor 1 IL-7Ralpha TM mut20 CD28 intracellular +4-1BB intracellular SEQ ID NO:146 SEQ ID NO:914
TGF-beta receptor 1 IL-7Ralpha TM mut21 CD28 intracellular +4-1BB intracellular SEQ ID NO:147 SEQ ID NO:915
TGF-beta receptor 1 IL-7Ralpha TM mut22 CD28 intracellular +4-1BB intracellular SEQ ID NO:148 SEQ ID NO:916
TGF-beta receptor 1 IL-7Ralpha TM mut23 CD28 intracellular +4-1BB intracellular SEQ ID NO:149 SEQ ID NO:917
TGF-beta receptor 1 IL-7Ralpha TM mut24 CD28 intracellular +4-1BB intracellular SEQ ID NO:150 SEQ ID NO:918
TGF-beta receptor 1 IL-7Ralpha TM mut25 CD28 intracellular +4-1BB intracellular SEQ ID NO:151 SEQ ID NO:919
TGF-beta receptor 1 IL-7Ralpha TM mut26 CD28 intracellular +4-1BB intracellular SEQ ID NO:152 SEQ ID NO:920
TGF-beta receptor 1 IL-7Ralpha TM mut27 CD28 intracellular +4-1BB intracellular SEQ ID NO:153 SEQ ID NO:921
TGF-beta receptor 1 IL-7Ralpha TM mut28 CD28 intracellular +4-1BB intracellular SEQ ID NO:154 SEQ ID NO:922
TGF-beta receptor 1 IL-7Ralpha TM mut29 CD28 intracellular +4-1BB intracellular SEQ ID NO:155 SEQ ID NO:923
TGF-beta receptor 1 IL-7Ralpha TM mut30 CD28 intracellular +4-1BB intracellular SEQ ID NO:156 SEQ ID NO:924
TGF-beta receptor 1 IL-7Ralpha TM mut31 CD28 intracellular +4-1BB intracellular SEQ ID NO:157 SEQ ID NO:925
TGF-beta receptor 1 IL-7Ralpha TM mut32 CD28 intracellular +4-1BB intracellular SEQ ID NO:158 SEQ ID NO:926
TGF-beta receptor 1 IL-7Ralpha TM mut1 CD28 intracellular domain SEQ ID NO:159 SEQ ID NO:927
TGF-beta receptor 1 IL-7Ralpha TM mut2 CD28 intracellular domain SEQ ID NO:160 SEQ ID NO:928
TGF-beta receptor 1 IL-7Ralpha TM mut3 CD28 intracellular domain SEQ ID NO:161 SEQ ID NO:929
TGF-beta receptor 1 IL-7Ralpha TM mut4 CD28 intracellular domain SEQ ID NO:162 SEQ ID NO:930
TGF-beta receptor 1 IL-7Ralpha TM mut5 CD28 intracellular domain SEQ ID NO:163 SEQ ID NO:931
TGF-beta receptor 1 IL-7Ralpha TM mut6 CD28 intracellular domain SEQ ID NO:164 SEQ ID NO:932
TGF-beta receptor 1 IL-7Ralpha TM mut7 CD28 intracellular domain SEQ ID NO:165 SEQ ID NO:933
TGF-beta receptor 1 IL-7Ralpha TM mut8 CD28 intracellular domain SEQ ID NO:166 SEQ ID NO:934
TGF-beta receptor 1 IL-7Ralpha TM mut9 CD28 intracellular domain SEQ ID NO:167 SEQ ID NO:935
TGF-beta receptor 1 IL-7Ralpha TM mut10 CD28 intracellular domain SEQ ID NO:168 SEQ ID NO:936
TGF-beta receptor 1 IL-7Ralpha TM mut11 CD28 intracellular domain SEQ ID NO:169 SEQ ID NO:937
TGF-beta receptor 1 IL-7Ralpha TM mut12 CD28 intracellular domain SEQ ID NO:170 SEQ ID NO:938
TGF-beta receptor 1 IL-7Ralpha TM mut13 CD28 intracellular domain SEQ ID NO:171 SEQ ID NO:939
TGF-beta receptor 1 IL-7Ralpha TM mut14 CD28 intracellular domain SEQ ID NO:172 SEQ ID NO:940
TGF-beta receptor 1 IL-7Ralpha TM mut15 CD28 intracellular domain SEQ ID NO:173 SEQ ID NO:941
TGF-beta receptor 1 IL-7Ralpha TM mut16 CD28 intracellular domain SEQ ID NO:174 SEQ ID NO:942
TGF-beta receptor 1 IL-7Ralpha TM mut17 CD28 intracellular domain SEQ ID NO:175 SEQ ID NO:943
TGF-beta receptor 1 IL-7Ralpha TM mut18 CD28 intracellular domain SEQ ID NO:176 SEQ ID NO:944
TGF-beta receptor 1 IL-7Ralpha TM mut19 CD28 intracellular domain SEQ ID NO:177 SEQ ID NO:945
TGF-beta receptor 1 IL-7Ralpha TM mut20 CD28 intracellular domain SEQ ID NO:178 SEQ ID NO:946
TGF-beta receptor 1 IL-7Ralpha TM mut21 CD28 intracellular domain SEQ ID NO:179 SEQ ID NO:947
TGF-beta receptor 1 IL-7Ralpha TM mut22 CD28 intracellular domain SEQ ID NO:180 SEQ ID NO:948
TGF-beta receptor 1 IL-7Ralpha TM mut23 CD28 intracellular domain SEQ ID NO:181 SEQ ID NO:949
TGF-beta receptor 1 IL-7Ralpha TM mut24 CD28 intracellular domain SEQ ID NO:182 SEQ ID NO:950
TGF-beta receptor 1 IL-7Ralpha TM mut25 CD28 intracellular domain SEQ ID NO:183 SEQ ID NO:951
TGF-beta receptor 1 IL-7Ralpha TM mut26 CD28 intracellular domain SEQ ID NO:184 SEQ ID NO:952
TGF-beta receptor 1 IL-7Ralpha TM mut27 CD28 intracellular domain SEQ ID NO:185 SEQ ID NO:953
TGF-beta receptor 1 IL-7Ralpha TM mut28 CD28 intracellular domain SEQ ID NO:186 SEQ ID NO:954
TGF-beta receptor 1 IL-7Ralpha TM mut29 CD28 intracellular domain SEQ ID NO:187 SEQ ID NO:955
TGF-beta receptor 1 IL-7Ralpha TM mut30 CD28 intracellular domain SEQ ID NO:188 SEQ ID NO:956
TGF-beta receptor 1 IL-7Ralpha TM mut31 CD28 intracellular domain SEQ ID NO:189 SEQ ID NO:957
TGF-beta receptor 1 IL-7Ralpha TM mut32 CD28 intracellular domain SEQ ID NO:190 SEQ ID NO:958
TGF-beta receptor 1 IL-7Ralpha TM mut1 CD28 intracellular + OX40 intracellular SEQ ID NO:191 SEQ ID NO:959
TGF-beta receptor 1 IL-7Ralpha TM mut2 CD28 intracellular + OX40 intracellular SEQ ID NO:192 SEQ ID NO:960
TGF-beta receptor 1 IL-7Ralpha TM mut3 CD28 intracellular + OX40 intracellular SEQ ID NO:193 SEQ ID NO:961
TGF-beta receptor 1 IL-7Ralpha TM mut4 CD28 intracellular + OX40 intracellular SEQ ID NO:194 SEQ ID NO:962
TGF-beta receptor 1 IL-7Ralpha TM mut5 CD28 intracellular + OX40 intracellular SEQ ID NO:195 SEQ ID NO:963
TGF-beta receptor 1 IL-7Ralpha TM mut6 CD28 intracellular + OX40 intracellular SEQ ID NO:196 SEQ ID NO:964
TGF-beta receptor 1 IL-7Ralpha TM mut7 CD28 intracellular + OX40 intracellular SEQ ID NO:197 SEQ ID NO:965
TGF-beta receptor 1 IL-7Ralpha TM mut8 CD28 intracellular + OX40 intracellular SEQ ID NO:198 SEQ ID NO:966
TGF-beta receptor 1 IL-7Ralpha TM mut9 CD28 intracellular + OX40 intracellular SEQ ID NO:199 SEQ ID NO:967
TGF-beta receptor 1 IL-7Ralpha TM mut10 CD28 intracellular + OX40 intracellular SEQ ID NO:200 SEQ ID NO:968
TGF-beta receptor 1 IL-7Ralpha TM mut11 CD28 intracellular + OX40 intracellular SEQ ID NO:201 SEQ ID NO:969
TGF-beta receptor 1 IL-7Ralpha TM mut12 CD28 intracellular + OX40 intracellular SEQ ID NO:202 SEQ ID NO:970
TGF-beta receptor 1 IL-7Ralpha TM mut13 CD28 intracellular + OX40 intracellular SEQ ID NO:203 SEQ ID NO:971
TGF-beta receptor 1 IL-7Ralpha TM mut14 CD28 intracellular + OX40 intracellular SEQ ID NO:204 SEQ ID NO:972
TGF-beta receptor 1 IL-7Ralpha TM mut15 CD28 intracellular + OX40 intracellular SEQ ID NO:205 SEQ ID NO:973
TGF-beta receptor 1 IL-7Ralpha TM mut16 CD28 intracellular + OX40 intracellular SEQ ID NO:206 SEQ ID NO:974
TGF-beta receptor 1 IL-7Ralpha TM mut17 CD28 intracellular + OX40 intracellular SEQ ID NO:207 SEQ ID NO:975
TGF-beta receptor 1 IL-7Ralpha TM mut18 CD28 intracellular + OX40 intracellular SEQ ID NO:208 SEQ ID NO:976
TGF-beta receptor 1 IL-7Ralpha TM mut19 CD28 intracellular + OX40 intracellular SEQ ID NO:209 SEQ ID NO:977
TGF-beta receptor 1 IL-7Ralpha TM mut20 CD28 intracellular + OX40 intracellular SEQ ID NO:210 SEQ ID NO:978
TGF-beta receptor 1 IL-7Ralpha TM mut21 CD28 intracellular + OX40 intracellular SEQ ID NO:211 SEQ ID NO:979
TGF-beta receptor 1 IL-7Ralpha TM mut22 CD28 intracellular + OX40 intracellular SEQ ID NO:212 SEQ ID NO:980
TGF-beta receptor 1 IL-7Ralpha TM mut23 CD28 intracellular + OX40 intracellular SEQ ID NO:213 SEQ ID NO:981
TGF-beta receptor 1 IL-7Ralpha TM mut24 CD28 intracellular + OX40 intracellular SEQ ID NO:214 SEQ ID NO:982
TGF-beta receptor 1 IL-7Ralpha TM mut25 CD28 intracellular + OX40 intracellular SEQ ID NO:215 SEQ ID NO:983
TGF-beta receptor 1 IL-7Ralpha TM mut26 CD28 intracellular + OX40 intracellular SEQ ID NO:216 SEQ ID NO:984
TGF-beta receptor 1 IL-7Ralpha TM mut27 CD28 intracellular + OX40 intracellular SEQ ID NO:217 SEQ ID NO:985
TGF-beta receptor 1 IL-7Ralpha TM mut28 CD28 intracellular + OX40 intracellular SEQ ID NO:218 SEQ ID NO:986
TGF-beta receptor 1 IL-7Ralpha TM mut29 CD28 intracellular + OX40 intracellular SEQ ID NO:219 SEQ ID NO:987
TGF-beta receptor 1 IL-7Ralpha TM mut30 CD28 intracellular + OX40 intracellular SEQ ID NO:220 SEQ ID NO:988
TGF-beta receptor 1 IL-7Ralpha TM mut31 CD28 intracellular + OX40 intracellular SEQ ID NO:221 SEQ ID NO:989
TGF-beta receptor 1 IL-7Ralpha TM mut32 CD28 intracellular + OX40 intracellular SEQ ID NO:222 SEQ ID NO:990
TGF-beta receptor 1 IL-7Ralpha TM mut1 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:223 SEQ ID NO:991
TGF-beta receptor 1 IL-7Ralpha TM mut2 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:224 SEQ ID NO:992
TGF-beta receptor 1 IL-7Ralpha TM mut3 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:225 SEQ ID NO:993
TGF-beta receptor 1 IL-7Ralpha TM mut4 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:226 SEQ ID NO:994
TGF-beta receptor 1 IL-7Ralpha TM mut5 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:227 SEQ ID NO:995
TGF-beta receptor 1 IL-7Ralpha TM mut6 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:228 SEQ ID NO:996
TGF-beta receptor 1 IL-7Ralpha TM mut7 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:229 SEQ ID NO:997
TGF-beta receptor 1 IL-7Ralpha TM mut8 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:230 SEQ ID NO:998
TGF-beta receptor 1 IL-7Ralpha TM mut9 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:231 SEQ ID NO:999
TGF-beta receptor 1 IL-7Ralpha TM mut10 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:232 SEQ ID NO:1000
TGF-beta receptor 1 IL-7Ralpha TM mut11 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:233 SEQ ID NO:1001
TGF-beta receptor 1 IL-7Ralpha TM mut12 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:234 SEQ ID NO:1002
TGF-beta receptor 1 IL-7Ralpha TM mut13 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:235 SEQ ID NO:1003
TGF-beta receptor 1 IL-7Ralpha TM mut14 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:236 SEQ ID NO:1004
TGF-beta receptor 1 IL-7Ralpha TM mut15 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:237 SEQ ID NO:1005
TGF-beta receptor 1 IL-7Ralpha TM mut16 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:238 SEQ ID NO:1006
TGF-beta receptor 1 IL-7Ralpha TM mut17 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:239 SEQ ID NO:1007
TGF-beta receptor 1 IL-7Ralpha TM mut18 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:240 SEQ ID NO:1008
TGF-beta receptor 1 IL-7Ralpha TM mut19 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:241 SEQ ID NO:1009
TGF-beta receptor 1 IL-7Ralpha TM mut20 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:242 SEQ ID NO:1010
TGF-beta receptor 1 IL-7Ralpha TM mut21 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:243 SEQ ID NO:1011
TGF-beta receptor 1 IL-7Ralpha TM mut22 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:244 SEQ ID NO:1012
TGF-beta receptor 1 IL-7Ralpha TM mut23 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:245 SEQ ID NO:1013
TGF-beta receptor 1 IL-7Ralpha TM mut24 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:246 SEQ ID NO:1014
TGF-beta receptor 1 IL-7Ralpha TM mut25 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:247 SEQ ID NO:1015
TGF-beta receptor 1 IL-7Ralpha TM mut26 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:248 SEQ ID NO:1016
TGF-beta receptor 1 IL-7Ralpha TM mut27 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:249 SEQ ID NO:1017
TGF-beta receptor 1 IL-7Ralpha TM mut28 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:250 SEQ ID NO:1018
TGF-beta receptor 1 IL-7Ralpha TM mut29 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:251 SEQ ID NO:1019
TGF-beta receptor 1 IL-7Ralpha TM mut30 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:252 SEQ ID NO:1020
TGF-beta receptor 1 IL-7Ralpha TM mut31 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:253 SEQ ID NO:1021
TGF-beta receptor 1 IL-7Ralpha TM mut32 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:254 SEQ ID NO:1022
TGF-beta receptor 1 IL-7Ralpha TM mut1 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:255 SEQ ID NO:1023
TGF-beta receptor 1 IL-7Ralpha TM mut2 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:256 SEQ ID NO:1024
TGF-beta receptor 1 IL-7Ralpha TM mut3 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:257 SEQ ID NO:1025
TGF-beta receptor 1 IL-7Ralpha TM mut4 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:258 SEQ ID NO:1026
TGF-beta receptor 1 IL-7Ralpha TM mut5 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:259 SEQ ID NO:1027
TGF-beta receptor 1 IL-7Ralpha TM mut6 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:260 SEQ ID NO:1028
TGF-beta receptor 1 IL-7Ralpha TM mut7 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:261 SEQ ID NO:1029
TGF-beta receptor 1 IL-7Ralpha TM mut8 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:262 SEQ ID NO:1030
TGF-beta receptor 1 IL-7Ralpha TM mut9 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:263 SEQ ID NO:1031
TGF-beta receptor 1 IL-7Ralpha TM mut10 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:264 SEQ ID NO:1032
TGF-beta receptor 1 IL-7Ralpha TM mut11 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:265 SEQ ID NO:1033
TGF-beta receptor 1 IL-7Ralpha TM mut12 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:266 SEQ ID NO:1034
TGF-beta receptor 1 IL-7Ralpha TM mut13 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:267 SEQ ID NO:1035
TGF-beta receptor 1 IL-7Ralpha TM mut14 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:268 SEQ ID NO:1036
TGF-beta receptor 1 IL-7Ralpha TM mut15 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:269 SEQ ID NO:1037
TGF-beta receptor 1 IL-7Ralpha TM mut16 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:270 SEQ ID NO:1038
TGF-beta receptor 1 IL-7Ralpha TM mut17 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:271 SEQ ID NO:1039
TGF-beta receptor 1 IL-7Ralpha TM mut18 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:272 SEQ ID NO:1040
TGF-beta receptor 1 IL-7Ralpha TM mut19 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:273 SEQ ID NO:1041
TGF-beta receptor 1 IL-7Ralpha TM mut20 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:274 SEQ ID NO:1042
TGF-beta receptor 1 IL-7Ralpha TM mut21 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:275 SEQ ID NO:1043
TGF-beta receptor 1 IL-7Ralpha TM mut22 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:276 SEQ ID NO:1044
TGF-beta receptor 1 IL-7Ralpha TM mut23 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:277 SEQ ID NO:1045
TGF-beta receptor 1 IL-7Ralpha TM mut24 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:278 SEQ ID NO:1046
TGF-beta receptor 1 IL-7Ralpha TM mut25 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:279 SEQ ID NO:1047
TGF-beta receptor 1 IL-7Ralpha TM mut26 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:280 SEQ ID NO:1048
TGF-beta receptor 1 IL-7Ralpha TM mut27 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:281 SEQ ID NO:1049
TGF-beta receptor 1 IL-7Ralpha TM mut28 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:282 SEQ ID NO:1050
TGF-beta receptor 1 IL-7Ralpha TM mut29 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:283 SEQ ID NO:1051
TGF-beta receptor 1 IL-7Ralpha TM mut30 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:284 SEQ ID NO:1052
TGF-beta receptor 1 IL-7Ralpha TM mut31 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:285 SEQ ID NO:1053
TGF-beta receptor 1 IL-7Ralpha TM mut32 28-DThe intracellular region of eltaIL2RB-z (YXXQ) SEQ ID NO:286 SEQ ID NO:1054
TGF-beta receptor 1 IL-7Ralpha TM mut1 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:287 SEQ ID NO:1055
TGF-beta receptor 1 IL-7Ralpha TM mut2 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:288 SEQ ID NO:1056
TGF-beta receptor 1 IL-7Ralpha TM mut3 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:289 SEQ ID NO:1057
TGF-beta receptor 1 IL-7Ralpha TM mut4 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:290 SEQ ID NO:1058
TGF-beta receptor 1 IL-7Ralpha TM mut5 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:291 SEQ ID NO:1059
TGF-beta receptorBody 1 IL-7Ralpha TM mut6 IL2RB-z (YXXQ) intracellular region SEQ ID NO:292 SEQ ID NO:1060
TGF-beta receptor 1 IL-7Ralpha TM mut7 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:293 SEQ ID NO:1061
TGF-beta receptor 1 IL-7Ralpha TM mut8 IL2RB-z (YXXQ) intracellular region SEQ ID NO:294 SEQ ID NO:1062
TGF-beta receptor 1 IL-7Ralpha TM mut9 IL2RB-z (YXXQ) intracellular region SEQ ID NO:295 SEQ ID NO:1063
TGF-beta receptor 1 IL-7Ralpha TM mut10 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:296 SEQ ID NO:1064
TGF-beta receptor 1 IL-7Ralpha TM mut11 IL2RB-z (YXXQ) intracellular region SEQ ID NO:297 SEQ ID NO:1065
TGF-beta receptor 1 IL-7Ralpha TM mut12 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:298 SEQ ID NO:1066
TGF-beta receptor 1 IL-7Ralpha TM mut13 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:299 SEQ ID NO:1067
TGF-beta receptor 1 IL-7Ralpha TM mut14 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:300 SEQ ID NO:1068
TGF-beta receptor 1 IL-7Ralpha TM mut15 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:301 SEQ ID NO:1069
TGF-beta receptor 1 IL-7Ralpha TM mut16 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:302 SEQ ID NO:1070
TGF-beta receptor 1 IL-7Ralpha TM mut17 IL2RB-z(YXXQ) intracellular region SEQ ID NO:303 SEQ ID NO:1071
TGF-beta receptor 1 IL-7Ralpha TM mut18 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:304 SEQ ID NO:1072
TGF-beta receptor 1 IL-7Ralpha TM mut19 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:305 SEQ ID NO:1073
TGF-beta receptor 1 IL-7Ralpha TM mut20 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:306 SEQ ID NO:1074
TGF-beta receptor 1 IL-7Ralpha TM mut21 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:307 SEQ ID NO:1075
TGF-beta receptor 1 IL-7Ralpha TM mut22 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:308 SEQ ID NO:1076
TGF-beta receptor 1 IL-7Ralpha TM mut23 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:309 SEQ ID NO:1077
TGF-beta receptor 1 IL-7Ralpha TM mut24 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:310 SEQ ID NO:1078
TGF-beta receptor 1 IL-7Ralpha TM mut25 IL2RB-z (YXXQ) intracellular region SEQ ID NO:311 SEQ ID NO:1079
TGF-beta receptor 1 IL-7Ralpha TM mut26 IL2RB-z (YXXQ) intracellular region SEQ ID NO:312 SEQ ID NO:1080
TGF-beta receptor 1 IL-7Ralpha TM mut27 IL2RB-z (YXXQ) intracellular region SEQ ID NO:313 SEQ ID NO:1081
TGF-beta receptor 1 IL-7Ralpha TM mut28 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:314 SEQ ID NO:1082
TGF-beta receptor 1 IL-7Ralpha TM mut29 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:315 SEQ ID NO:1083
TGF-beta receptor 1 IL-7Ralpha TM mut30 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:316 SEQ ID NO:1084
TGF-beta receptor 1 IL-7Ralpha TM mut31 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:317 SEQ ID NO:1085
TGF-beta receptor 1 IL-7Ralpha TM mut32 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:318 SEQ ID NO:1086
TGF-beta receptor 1 IL-7Ralpha TM mut1 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:319 SEQ ID NO:1087
TGF-beta receptor 1 IL-7Ralpha TM mut2 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:320 SEQ ID NO:1088
TGF-beta receptor 1 IL-7Ralpha TM mut3 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:321 SEQ ID NO:1089
TGF-beta receptor 1 IL-7Ralpha TM mut4 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:322 SEQ ID NO:1090
TGF-beta receptor 1 IL-7Ralpha TM mut5 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:323 SEQ ID NO:1091
TGF-beta receptor 1 IL-7Ralpha TM mut6 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:324 SEQ ID NO:1092
TGF-beta receptor 1 IL-7Ralpha TM mut7 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:325 SEQ ID NO:1093
TGF-beta receptor 1 IL-7Ralpha TM mut8 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:326 SEQ ID NO:1094
TGF-beta receptor 1 IL-7Ralpha TM mut9 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:327 SEQ ID NO:1095
TGF-beta receptor 1 IL-7Ralpha TM mut10 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:328 SEQ ID NO:1096
TGF-beta receptor 1 IL-7Ralpha TM mut11 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:329 SEQ ID NO:1097
TGF-beta receptor 1 IL-7Ralpha TM mut12 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:330 SEQ ID NO:1098
TGF-beta receptor 1 IL-7Ralpha TM mut13 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:331 SEQ ID NO:1099
TGF-beta receptor 1 IL-7Ralpha TM mut14 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:332 SEQ ID NO:1100
TGF-beta receptor 1 IL-7Ralpha TM mut15 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:333 SEQ ID NO:1101
TGF-beta receptor 1 IL-7Ralpha TM mut16 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:334 SEQ ID NO:1102
TGF-beta receptor 1 IL-7Ralpha TM mut17 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:335 SEQ ID NO:1103
TGF-beta receptor 1 IL-7Ralpha TM mut18 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:336 SEQ ID NO:1104
TGF-beta receptor 1 IL-7Ralpha TM mut19 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:337 SEQ ID NO:1105
TGF-beta receptor 1 IL-7Ralpha TM mut20 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:338 SEQ ID NO:1106
TGF-beta receptor 1 IL-7Ralpha TM mut21 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:339 SEQ ID NO:1107
TGF-beta receptor 1 IL-7Ralpha TM mut22 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:340 SEQ ID NO:1108
TGF-beta receptor 1 IL-7Ralpha TM mut23 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:341 SEQ ID NO:1109
TGF-beta receptor 1 IL-7Ralpha TM mut24 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:342 SEQ ID NO:1110
TGF-beta receptor 1 IL-7Ralpha TM mut25 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:343 SEQ ID NO:1111
TGF-beta receptor 1 IL-7Ralpha TM mut26 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:344 SEQ ID NO:1112
TGF-beta receptor 1 IL-7Ralpha TM mut27 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:345 SEQ ID NO:1113
TGF-beta receptor 1 IL-7Ralpha TM mut28 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:346 SEQ ID NO:1114
TGF-beta receptor 1 IL-7Ralpha TM mut29 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:347 SEQ ID NO:1115
TGF-beta receptor 1 IL-7Ralpha TM mut30 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:348 SEQ ID NO:1116
TGF-beta receptor 1 IL-7Ralpha TM mut31 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:349 SEQ ID NO:1117
TGF-beta receptor 1 IL-7Ralpha TM mut32 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:350 SEQ ID NO:1118
TGF-beta receptor 2 IL-7Ralpha TM mut1 Intracellular domain of IL-7Ralpha SEQ ID NO:351 SEQ ID NO:1119
TGF-beta receptor 2 IL-7Ralpha TM mut2 Intracellular domain of IL-7Ralpha SEQ ID NO:352 SEQ ID NO:1120
TGF-beta receptor 2 IL-7Ralpha TM mut3 Intracellular domain of IL-7Ralpha SEQ ID NO:353 SEQ ID NO:1121
TGF-beta receptor 2 IL-7Ralpha TM mut4 Intracellular domain of IL-7Ralpha SEQ ID NO:354 SEQ ID NO:1122
TGF-beta receptor 2 IL-7Ralpha TM mut5 Intracellular domain of IL-7Ralpha SEQ ID NO:355 SEQ ID NO:1123
TGF-beta receptor 2 IL-7Ralpha TM mut6 Intracellular domain of IL-7Ralpha SEQ ID NO:356 SEQ ID NO:1124
TGF-beta receptor 2 IL-7Ralpha TM mut7 Intracellular domain of IL-7Ralpha SEQ ID NO:357 SEQ ID NO:1125
TGF-beta receptor 2 IL-7Ralpha TM mut8 Intracellular domain of IL-7Ralpha SEQ ID NO:358 SEQ ID NO:1126
TGF-beta receptor 2 IL-7Ralpha TM mut9 Intracellular domain of IL-7Ralpha SEQ ID NO:359 SEQ ID NO:1127
TGF-beta receptor 2 IL-7Ralpha TM mut10 Intracellular domain of IL-7Ralpha SEQ ID NO:360 SEQ ID NO:1128
TGF-beta receptor 2 IL-7Ralpha TM mut11 Intracellular domain of IL-7Ralpha SEQ ID NO:361 SEQ ID NO:1129
TGF-beta receptor 2 IL-7Ralpha TM mut12 Intracellular domain of IL-7Ralpha SEQ ID NO:362 SEQ ID NO:1130
TGF-beta receptor 2 IL-7Ralpha TM mut13 Intracellular domain of IL-7Ralpha SEQ ID NO:363 SEQ ID NO:1131
TGF-beta receptor 2 IL-7Ralpha TM mut14 Intracellular domain of IL-7Ralpha SEQ ID NO:364 SEQ ID NO:1132
TGF-beta receptor 2 IL-7Ralpha TM mut15 Intracellular domain of IL-7Ralpha SEQ ID NO:365 SEQ ID NO:1133
TGF-beta receptor 2 IL-7Ralpha TM mut16 Intracellular domain of IL-7Ralpha SEQ ID NO:366 SEQ ID NO:1134
TGF-beta receptor 2 IL-7Ralpha TM mut17 Intracellular domain of IL-7Ralpha SEQ ID NO:367 SEQ ID NO:1135
TGF-beta receptor 2 IL-7Ralpha TM mut18 Intracellular domain of IL-7Ralpha SEQ ID NO:368 SEQ ID NO:1136
TGF-beta receptor 2 IL-7Ralpha TM mut19 Intracellular domain of IL-7Ralpha SEQ ID NO:369 SEQ ID NO:1137
TGF-beta receptor 2 IL-7Ralpha TM mut20 Intracellular domain of IL-7Ralpha SEQ ID NO:370 SEQ ID NO:1138
TGF-beta receptor 2 IL-7Ralpha TM mut21 Intracellular domain of IL-7Ralpha SEQ ID NO:371 SEQ ID NO:1139
TGF-beta receptor 2 IL-7Ralpha TM mut22 Intracellular domain of IL-7Ralpha SEQ ID NO:372 SEQ ID NO:1140
TGF-beta receptor 2 IL-7Ralpha TM mut23 Intracellular domain of IL-7Ralpha SEQ ID NO:373 SEQ ID NO:1141
TGF-beta receptor 2 IL-7Ralpha TM mut24 Intracellular domain of IL-7Ralpha SEQ ID NO:374 SEQ ID NO:1142
TGF-beta receptor 2 IL-7Ralpha TM mut25 Intracellular domain of IL-7Ralpha SEQ ID NO:375 SEQ ID NO:1143
TGF-beta receptor 2 IL-7Ralpha TM mut26 Intracellular domain of IL-7Ralpha SEQ ID NO:376 SEQ ID NO:1144
TGF-beta receptor 2 IL-7Ralpha TM mut27 Intracellular domain of IL-7Ralpha SEQ ID NO:377 SEQ ID NO:1145
TGF-beta receptor 2 IL-7Ralpha TM mut28 Intracellular domain of IL-7Ralpha SEQ ID NO:378 SEQ ID NO:1146
TGF-beta receptor 2 IL-7Ralpha TM mut29 Intracellular domain of IL-7Ralpha SEQ ID NO:379 SEQ ID NO:1147
TGF-beta receptor 2 IL-7Ralpha TM mut30 Intracellular domain of IL-7Ralpha SEQ ID NO:380 SEQ ID NO:1148
TGF-beta receptor 2 IL-7Ralpha TM mut31 Intracellular domain of IL-7Ralpha SEQ ID NO:381 SEQ ID NO:1149
TGF-beta receptor 2 IL-7Ralpha TM mut32 Intracellular domain of IL-7Ralpha SEQ ID NO:382 SEQ ID NO:1150
TGF-beta receptor 2 IL-7Ralpha TM mut1 CD28 intracellular +4-1BB intracellular SEQ ID NO:383 SEQ ID NO:1151
TGF-beta receptor 2 IL-7Ralpha TM mut2 CD28 intracellular +4-1BB intracellular SEQ ID NO:384 SEQ ID NO:1152
TGF-beta receptor 2 IL-7Ralpha TM mut3 CD28 intracellular +4-1BB intracellular SEQ ID NO:385 SEQ ID NO:1153
TGF-beta receptor 2 IL-7Ralpha TM mut4 CD28 intracellular +4-1BB intracellular SEQ ID NO:386 SEQ ID NO:1154
TGF-beta receptor 2 IL-7Ralpha TM mut5 CD28 intracellular +4-1BB intracellular SEQ ID NO:387 SEQ ID NO:1155
TGF-beta receptor 2 IL-7Ralpha TM mut6 CD28 intracellular +4-1BB intracellular SEQ ID NO:388 SEQ ID NO:1156
TGF-beta receptor 2 IL-7Ralpha TM mut7 CD28 intracellular +4-1BB intracellular SEQ ID NO:389 SEQ ID NO:1157
TGF-beta receptor 2 IL-7Ralpha TM mut8 CD28 intracellular +4-1BB intracellular SEQ ID NO:390 SEQ ID NO:1158
TGF-beta receptor 2 IL-7Ralpha TM mut9 CD28 intracellular +4-1BB intracellular SEQ ID NO:391 SEQ ID NO:1159
TGF-beta receptor 2 IL-7Ralpha TM mut10 CD28 intracellular +4-1BB intracellular SEQ ID NO:392 SEQ ID NO:1160
TGF-beta receptor 2 IL-7Ralpha TM mut11 CD28 intracellular +4-1BB intracellular SEQ ID NO:393 SEQ ID NO:1161
TGF-beta receptor 2 IL-7Ralpha TM mut12 CD28 intracellular +4-1BB intracellular SEQ ID NO:394 SEQ ID NO:1162
TGF-beta receptor 2 IL-7Ralpha TM mut13 CD28 intracellular +4-1BB intracellular SEQ ID NO:395 SEQ ID NO:1163
TGF-beta receptor 2 IL-7Ralpha TM mut14 CD28 intracellular +4-1BB intracellular region SEQ ID NO:396 SEQ ID NO:1164
TGF-beta receptor 2 IL-7Ralpha TM mut15 CD28 intracellular +4-1BB intracellular SEQ ID NO:397 SEQ ID NO:1165
TGF-beta receptor 2 IL-7Ralpha TM mut16 CD28 intracellular +4-1BB intracellular SEQ ID NO:398 SEQ ID NO:1166
TGF-beta receptor 2 IL-7Ralpha TM mut17 CD28 intracellular +4-1BB intracellular SEQ ID NO:399 SEQ ID NO:1167
TGF-beta receptor 2 IL-7Ralpha TM mut18 CD28 intracellular +4-1BB intracellular SEQ ID NO:400 SEQ ID NO:1168
TGF-beta receptor 2 IL-7Ralpha TM mut19 CD28 intracellular +4-1BB intracellular SEQ ID NO:401 SEQ ID NO:1169
TGF-beta receptor 2 IL-7Ralpha TM mut20 CD28 intracellular +4-1BB intracellular SEQ ID NO:402 SEQ ID NO:1170
TGF-beta receptor 2 IL-7Ralpha TM mut21 CD28 intracellular +4-1BB intracellular SEQ ID NO:403 SEQ ID NO:1171
TGF-beta receptor 2 IL-7Ralpha TM mut22 CD28 intracellular +4-1BB intracellular SEQ ID NO:404 SEQ ID NO:1172
TGF-beta receptor 2 IL-7Ralpha TM mut23 CD28 intracellular +4-1BB intracellular SEQ ID NO:405 SEQ ID NO:1173
TGF-beta receptor 2 IL-7Ralpha TM mut24 CD28 intracellular +4-1BB intracellular SEQ ID NO:406 SEQ ID NO:1174
TGF-beta receptor 2 IL-7Ralpha TM mut25 CD28 intracellular +4-1BB intracellular SEQ ID NO:407 SEQ ID NO:1175
TGF-beta receptor 2 IL-7Ralpha TM mut26 CD28 intracellular +4-1BB intracellular SEQ ID NO:408 SEQ ID NO:1176
TGF-beta receptor 2 IL-7Ralpha TM mut27 CD28 intracellular +4-1BB intracellular SEQ ID NO:409 SEQ ID NO:1177
TGF-beta receptor 2 IL-7Ralpha TM mut28 CD28 intracellular +4-1BB intracellular SEQ ID NO:410 SEQ ID NO:1178
TGF-beta receptor 2 IL-7Ralpha TM mut29 CD28 intracellular +4-1BB intracellular SEQ ID NO:411 SEQ ID NO:1179
TGF-beta receptor 2 IL-7Ralpha TM mut30 CD28 intracellular +4-1BB intracellular SEQ ID NO:412 SEQ ID NO:1180
TGF-beta receptor 2 IL-7Ralpha TM mut31 CD28 intracellular +4-1BB intracellular SEQ ID NO:413 SEQ ID NO:1181
TGF-beta receptor 2 IL-7Ralpha TM mut32 CD28 intracellular +4-1BB intracellular SEQ ID NO:414 SEQ ID NO:1182
TGF-beta receptor 2 IL-7Ralpha TM mut1 CD28 intracellular domain SEQ ID NO:415 SEQ ID NO:1183
TGF-beta receptor 2 IL-7Ralpha TM mut2 CD28 intracellular domain SEQ ID NO:416 SEQ ID NO:1184
TGF-beta receptor 2 IL-7Ralpha TM mut3 CD28 intracellular domain SEQ ID NO:417 SEQ ID NO:1185
TGF-beta receptor 2 IL-7Ralpha TM mut4 CD28 intracellular domain SEQ ID NO:418 SEQ ID NO:1186
TGF-beta receptor 2 IL-7Ralpha TM mut5 CD28 intracellular domain SEQ ID NO:419 SEQ ID NO:1187
TGF-beta receptor 2 IL-7Ralpha TM mut6 CD28 intracellular domain SEQ ID NO:420 SEQ ID NO:1188
TGF-beta receptor 2 IL-7Ralpha TM mut7 CD28 intracellular domain SEQ ID NO:421 SEQ ID NO:1189
TGF-beta receptor 2 IL-7Ralpha TM mut8 CD28 intracellular domain SEQ ID NO:422 SEQ ID NO:1190
TGF-beta receptor 2 IL-7Ralpha TM mut9 CD28 intracellular domain SEQ ID NO:423 SEQ ID NO:1191
TGF-beta receptor 2 IL-7Ralpha TM mut10 CD28 intracellular domain SEQ ID NO:424 SEQ ID NO:1192
TGF-beta receptor 2 IL-7Ralpha TM mut11 CD28 intracellular domain SEQ ID NO:425 SEQ ID NO:1193
TGF-beta receptor 2 IL-7Ralpha TM mut12 CD28 intracellular domain SEQ ID NO:426 SEQ ID NO:1194
TGF-beta receptor 2 IL-7Ralpha TM mut13 CD28 intracellular domain SEQ ID NO:427 SEQ ID NO:1195
TGF-beta receptor 2 IL-7Ralpha TM mut14 CD28 intracellular domain SEQ ID NO:428 SEQ ID NO:1196
TGF-beta receptor 2 IL-7Ralpha TM mut15 CD28 intracellular domain SEQ ID NO:429 SEQ ID NO:1197
TGF-beta receptor 2 IL-7Ralpha TM mut16 CD28 intracellular domain SEQ ID NO:430 SEQ ID NO:1198
TGF-beta receptor 2 IL-7Ralpha TM mut17 CD28 intracellular domain SEQ ID NO:431 SEQ ID NO:1199
TGF-beta receptor 2 IL-7Ralpha TM mut18 CD28 intracellular domain SEQ ID NO:432 SEQ ID NO:1200
TGF-beta receptor 2 IL-7Ralpha TM mut19 CD28 intracellular domain SEQ ID NO:433 SEQ ID NO:1201
TGF-beta receptor 2 IL-7Ralpha TM mut20 CD28 intracellular domain SEQ ID NO:434 SEQ ID NO:1202
TGF-beta receptor 2 IL-7Ralpha TM mut21 CD28 intracellular domain SEQ ID NO:435 SEQ ID NO:1203
TGF-beta receptor 2 IL-7Ralpha TM mut22 CD28 intracellular domain SEQ ID NO:436 SEQ ID NO:1204
TGF-beta receptor 2 IL-7Ralpha TM mut23 CD28 intracellular domain SEQ ID NO:437 SEQ ID NO:1205
TGF-beta receptor 2 IL-7Ralpha TM mut24 CD28 intracellular domain SEQ ID NO:438 SEQ ID NO:1206
TGF-beta receptor 2 IL-7Ralpha TM mut25 CD28 intracellular domain SEQ ID NO:439 SEQ ID NO:1207
TGF-beta receptor 2 IL-7Ralpha TM mut26 CD28 intracellular domain SEQ ID NO:440 SEQ ID NO:1208
TGF-beta receptor 2 IL-7Ralpha TM mut27 CD28 intracellular domain SEQ ID NO:441 SEQ ID NO:1209
TGF-beta receptor 2 IL-7Ralpha TM mut28 CD28 intracellular domain SEQ ID NO:442 SEQ ID NO:1210
TGF-beta receptor 2 IL-7Ralpha TM mut29 CD28 intracellular domain SEQ ID NO:443 SEQ ID NO:1211
TGF-beta receptor 2 IL-7Ralpha TM mut30 CD28 intracellular domain SEQ ID NO:444 SEQ ID NO:1212
TGF-beta receptor 2 IL-7Ralpha TM mut31 CD28 intracellular domain SEQ ID NO:445 SEQ ID NO:1213
TGF-beta receptor 2 IL-7Ralpha TM mut32 CD28 intracellular domain SEQ ID NO:446 SEQ ID NO:1214
TGF-beta receptor 2 IL-7Ralpha TM mut1 CD28 intracellular + OX40 intracellular SEQ ID NO:447 SEQ ID NO:1215
TGF-beta receptor 2 IL-7Ralpha TM mut2 CD28 intracellular + OX40 intracellular SEQ ID NO:448 SEQ ID NO:1216
TGF-beta receptor 2 IL-7Ralpha TM mut3 CD28 intracellular + OX40 intracellular SEQ ID NO:449 SEQ ID NO:1217
TGF-beta receptor 2 IL-7Ralpha TM mut4 CD28 intracellular + OX40 intracellular SEQ ID NO:450 SEQ ID NO:1218
TGF-beta receptor 2 IL-7Ralpha TM mut5 CD28 intracellular + OX40 intracellular SEQ ID NO:451 SEQ ID NO:1219
TGF-beta receptor 2 IL-7Ralpha TM mut6 CD28 intracellular + OX40 intracellular SEQ ID NO:452 SEQ ID NO:1220
TGF-beta receptor 2 IL-7Ralpha TM mut7 CD28 intracellular + OX40 intracellular SEQ ID NO:453 SEQ ID NO:1221
TGF-beta receptor 2 IL-7Ralpha TM mut8 CD28 intracellular + OX40 intracellular SEQ ID NO:454 SEQ ID NO:1222
TGF-beta receptor 2 IL-7Ralpha TM mut9 CD28 intracellular + OX40 intracellular SEQ ID NO:455 SEQ ID NO:1223
TGF-beta receptor2 IL-7Ralpha TM mut10 CD28 intracellular + OX40 intracellular SEQ ID NO:456 SEQ ID NO:1224
TGF-beta receptor 2 IL-7Ralpha TM mut11 CD28 intracellular + OX40 intracellular SEQ ID NO:457 SEQ ID NO:1225
TGF-beta receptor 2 IL-7Ralpha TM mut12 CD28 intracellular + OX40 intracellular SEQ ID NO:458 SEQ ID NO:1226
TGF-beta receptor 2 IL-7Ralpha TM mut13 CD28 intracellular + OX40 intracellular SEQ ID NO:459 SEQ ID NO:1227
TGF-beta receptor 2 IL-7Ralpha TM mut14 CD28 intracellular + OX40 intracellular SEQ ID NO:460 SEQ ID NO:1228
TGF-beta receptor 2 IL-7Ralpha TM mut15 CD28 intracellular + OX40 intracellular SEQ ID NO:461 SEQ ID NO:1229
TGF-beta receptor 2 IL-7Ralpha TM mut16 CD28 intracellular + OX40 intracellular SEQ ID NO:462 SEQ ID NO:1230
TGF-beta receptor 2 IL-7Ralpha TM mut17 CD28 intracellular + OX40 intracellular SEQ ID NO:463 SEQ ID NO:1231
TGF-beta receptor 2 IL-7Ralpha TM mut18 CD28 intracellular + OX40 intracellular SEQ ID NO:464 SEQ ID NO:1232
TGF-beta receptor 2 IL-7Ralpha TM mut19 CD28 intracellular + OX40 intracellular SEQ ID NO:465 SEQ ID NO:1233
TGF-beta receptor 2 IL-7Ralpha TM mut20 CD28 intracellular + OX40 intracellular SEQ ID NO:466 SEQ ID NO:1234
TGF-beta receptor 2 IL-7Ralpha TM mut21 CD28 intracellular + OX40 intracellular SEQ ID NO:467 SEQ ID NO:1235
TGF-beta receptor 2 IL-7Ralpha TM mut22 CD28 intracellular + OX40 intracellular SEQ ID NO:468 SEQ ID NO:1236
TGF-beta receptor 2 IL-7Ralpha TM mut23 CD28 intracellular + OX40 intracellular SEQ ID NO:469 SEQ ID NO:1237
TGF-beta receptor 2 IL-7Ralpha TM mut24 CD28 intracellular + OX40 intracellular SEQ ID NO:470 SEQ ID NO:1238
TGF-beta receptor 2 IL-7Ralpha TM mut25 CD28 intracellular + OX40 intracellular SEQ ID NO:471 SEQ ID NO:1239
TGF-beta receptor 2 IL-7Ralpha TM mut26 CD28 intracellular + OX40 intracellular SEQ ID NO:472 SEQ ID NO:1240
TGF-beta receptor 2 IL-7Ralpha TM mut27 CD28 intracellular + OX40 intracellular SEQ ID NO:473 SEQ ID NO:1241
TGF-beta receptor 2 IL-7Ralpha TM mut28 CD28 intracellular + OX40 intracellular SEQ ID NO:474 SEQ ID NO:1242
TGF-beta receptor 2 IL-7Ralpha TM mut29 CD28 intracellular + OX40 intracellular SEQ ID NO:475 SEQ ID NO:1243
TGF-beta receptor 2 IL-7Ralpha TM mut30 CD28 intracellular + OX40 intracellular SEQ ID NO:476 SEQ ID NO:1244
TGF-beta receptor 2 IL-7Ralpha TM mut31 CD28 intracellular + OX40 intracellular SEQ ID NO:477 SEQ ID NO:1245
TGF-beta receptor 2 IL-7Ralpha TM mut32 CD28 intracellular + OX40 intracellular SEQ ID NO:478 SEQ ID NO:1246
TGF-beta receptor 2 IL-7Ralpha TM mut1 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:479 SEQ ID NO:1247
TGF-beta receptor 2 IL-7Ralpha TM mut2 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:480 SEQ ID NO:1248
TGF-beta receptor 2 IL-7Ralpha TM mut3 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:481 SEQ ID NO:1249
TGF-beta receptor 2 IL-7Ralpha TM mut4 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:482 SEQ ID NO:1250
TGF-beta receptor 2 IL-7Ralpha TM mut5 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:483 SEQ ID NO:1251
TGF-beta receptor 2 IL-7Ralpha TM mut6 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:484 SEQ ID NO:1252
TGF-beta receptor 2 IL-7Ralpha TM mut7 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:485 SEQ ID NO:1253
TGF-beta receptor 2 IL-7Ralpha TM mut8 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:486 SEQ ID NO:1254
TGF-beta receptor 2 IL-7Ralpha TM mut9 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:487 SEQ ID NO:1255
TGF-beta receptor 2 IL-7Ralpha TM mut10 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:488 SEQ ID NO:1256
TGF-beta receptor 2 IL-7Ralpha TM mut11 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:489 SEQ ID NO:1257
TGF-beta receptor 2 IL-7Ralpha TM mut12 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:490 SEQ ID NO:1258
TGF-beta receptor 2 IL-7Ralpha TM mut13 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:491 SEQ ID NO:1259
TGF-beta receptor 2 IL-7Ralpha TM mut14 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:492 SEQ ID NO:1260
TGF-beta receptor 2 IL-7Ralpha TM mut15 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:493 SEQ ID NO:1261
TGF-beta receptor 2 IL-7Ralpha TM mut16 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:494 SEQ ID NO:1262
TGF-beta receptor 2 IL-7Ralpha TM mut17 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:495 SEQ ID NO:1263
TGF-beta receptor 2 IL-7Ralpha TM mut18 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:496 SEQ ID NO:1264
TGF-beta receptor 2 IL-7Ralpha TM mut19 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:497 SEQ ID NO:1265
TGF-beta receptor 2 IL-7Ralpha TM mut20 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:498 SEQ ID NO:1266
TGF-beta receptor 2 IL-7Ralpha TM mut21 28-IL2RB-z (YXXQ) intracellularlyZone(s) SEQ ID NO:499 SEQ ID NO:1267
TGF-beta receptor 2 IL-7Ralpha TM mut22 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:500 SEQ ID NO:1268
TGF-beta receptor 2 IL-7Ralpha TM mut23 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:501 SEQ ID NO:1269
TGF-beta receptor 2 IL-7Ralpha TM mut24 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:502 SEQ ID NO:1270
TGF-beta receptor 2 IL-7Ralpha TM mut25 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:503 SEQ ID NO:1271
TGF-beta receptor 2 IL-7Ralpha TM mut26 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:504 SEQ ID NO:1272
TGF-beta receptor2 IL-7Ralpha TM mut27 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:505 SEQ ID NO:1273
TGF-beta receptor 2 IL-7Ralpha TM mut28 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:506 SEQ ID NO:1274
TGF-beta receptor 2 IL-7Ralpha TM mut29 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:507 SEQ ID NO:1275
TGF-beta receptor 2 IL-7Ralpha TM mut30 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:508 SEQ ID NO:1276
TGF-beta receptor 2 IL-7Ralpha TM mut31 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:509 SEQ ID NO:1277
TGF-beta receptor 2 IL-7Ralpha TM mut32 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:510 SEQ ID NO:1278
TGF-beta receptor 2 IL-7Ralpha TM mut1 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:511 SEQ ID NO:1279
TGF-beta receptor 2 IL-7Ralpha TM mut2 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:512 SEQ ID NO:1280
TGF-beta receptor 2 IL-7Ralpha TM mut3 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:513 SEQ ID NO:1281
TGF-beta receptor 2 IL-7Ralpha TM mut4 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:514 SEQ ID NO:1282
TGF-beta receptor 2 IL-7Ralpha TM mut5 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:515 SEQ ID NO:1283
TGF-beta receptor 2 IL-7Ralpha TM mut6 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:516 SEQ ID NO:1284
TGF-beta receptor 2 IL-7Ralpha TM mut7 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:517 SEQ ID NO:1285
TGF-beta receptor 2 IL-7Ralpha TM mut8 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:518 SEQ ID NO:1286
TGF-beta receptor 2 IL-7Ralpha TM mut9 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:519 SEQ ID NO:1287
TGF-beta receptor 2 IL-7Ralpha TM mut10 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:520 SEQ ID NO:1288
TGF-beta receptor 2 IL-7Ralpha TM mut11 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:521 SEQ ID NO:1289
TGF-beta receptor 2 IL-7Ralpha TM mut12 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:522 SEQ ID NO:1290
TGF-beta receptor 2 IL-7Ralpha TM mut13 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:523 SEQ ID NO:1291
TGF-beta receptor 2 IL-7Ralpha TM mut14 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:524 SEQ ID NO:1292
TGF-beta receptor 2 IL-7Ralpha TM mut15 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:525 SEQ ID NO:1293
TGF-beta receptor 2 IL-7Ralpha TM mut16 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:526 SEQ ID NO:1294
TGF-beta receptor 2 IL-7Ralpha TM mut17 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:527 SEQ ID NO:1295
TGF-beta receptor 2 IL-7Ralpha TM mut18 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:528 SEQ ID NO:1296
TGF-beta receptor 2 IL-7Ralpha TM mut19 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:529 SEQ ID NO:1297
TGF-beta receptor 2 IL-7Ralpha TM mut20 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:530 SEQ ID NO:1298
TGF-beta receptor 2 IL-7Ralpha TM mut21 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:531 SEQ ID NO:1299
TGF-beta receptor 2 IL-7Ralpha TM mut22 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:532 SEQ ID NO:1300
TGF-beta receptor 2 IL-7Ralpha TM mut23 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:533 SEQ ID NO:1301
TGF-beta receptor 2 IL-7Ralpha TM mut24 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:534 SEQ ID NO:1302
TGF-beta receptor 2 IL-7Ralpha TM mut25 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:535 SEQ ID NO:1303
TGF-beta receptor 2 IL-7Ralpha TM mut26 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:536 SEQ ID NO:1304
TGF-beta receptor 2 IL-7Ralpha TM mut27 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:537 SEQ ID NO:1305
TGF-beta receptor 2 IL-7Ralpha TM mut28 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:538 SEQ ID NO:1306
TGF-beta receptor 2 IL-7Ralpha TM mut29 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:539 SEQ ID NO:1307
TGF-beta receptor 2 IL-7Ralpha TM mut30 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:540 SEQ ID NO:1308
TGF-beta receptor 2 IL-7Ralpha TM mut31 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:541 SEQ ID NO:1309
TGF-beta receptor 2 IL-7Ralpha TM mut32 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:542 SEQ ID NO:1310
TGF-beta receptor 2 IL-7Ralpha TM mut1 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:543 SEQ ID NO:1311
TGF-beta receptor 2 IL-7Ralpha TM mut2 IL2RB-z (YXXQ) intracellular region SEQ ID NO:544 SEQ ID NO:1312
TGF-beta receptor 2 IL-7Ralpha TM mut3 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:545 SEQ ID NO:1313
TGF-beta receptor 2 IL-7Ralpha TM mut4 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:546 SEQ ID NO:1314
TGF-beta receptor 2 IL-7Ralpha TM mut5 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:547 SEQ ID NO:1315
TGF-beta receptor 2 IL-7Ralpha TM mut6 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:548 SEQ ID NO:1316
TGF-beta receptor 2 IL-7Ralpha TM mut7 IL2RB-z (YXXQ) intracellular region SEQ ID NO:549 SEQ ID NO:1317
TGF-beta receptor 2 IL-7Ralpha TM mut8 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:550 SEQ ID NO:1318
TGF-beta receptor 2 IL-7Ralpha TM mut9 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:551 SEQ ID NO:1319
TGF-beta receptor 2 IL-7Ralpha TM mut10 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:552 SEQ ID NO:1320
TGF-beta receptor 2 IL-7Ralpha TM mut11 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:553 SEQ ID NO:1321
TGF-beta receptor 2 IL-7Ralpha TM mut12 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:554 SEQ ID NO:1322
TGF-beta receptor 2 IL-7Ralpha TM mut13 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:555 SEQ ID NO:1323
TGF-beta receptor 2 IL-7Ralpha TM mut14 IL2RB-z (YXXQ) intracellular region SEQ ID NO:556 SEQ ID NO:1324
TGF-beta receptor 2 IL-7Ralpha TM mut15 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:557 SEQ ID NO:1325
TGF-beta receptor 2 IL-7Ralpha TM mut16 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:558 SEQ ID NO:1326
TGF-beta receptor 2 IL-7Ralpha TM mut17 IL2RB-z (YXXQ) intracellular region SEQ ID NO:559 SEQ ID NO:1327
TGF-beta receptor 2 IL-7Ralpha TM mut18 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:560 SEQ ID NO:1328
TGF-beta receptor 2 IL-7Ralpha TM mut19 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:561 SEQ ID NO:1329
TGF-beta receptor 2 IL-7Ralpha TM mut20 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:562 SEQ ID NO:1330
TGF-beta receptor 2 IL-7Ralpha TM mut21 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:563 SEQ ID NO:1331
TGF-beta receptor 2 IL-7Ralpha TM mut22 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:564 SEQ ID NO:1332
TGF-beta receptor 2 IL-7Ralpha TM mut23 IL2RB-z (YXXQ) intracellular region SEQ ID NO:565 SEQ ID NO:1333
TGF-beta receptor 2 IL-7Ralpha TM mut24 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:566 SEQ ID NO:1334
TGF-beta receptor 2 IL-7Ralpha TM mut25 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:567 SEQ ID NO:1335
TGF-beta receptor 2 IL-7Ralpha TM mut26 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:568 SEQ ID NO:1336
TGF-beta receptor 2 IL-7Ralpha TM mut27 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:569 SEQ ID NO:1337
TGF-beta receptor 2 IL-7Ralpha TM mut28 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:570 SEQ ID NO:1338
TGF-beta receptor 2 IL-7Ralpha TM mut29 IL2RB-z (YXXQ) intracellular region SEQ ID NO:571 SEQ ID NO:1339
TGF-beta receptor 2 IL-7Ralpha TM mut30 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:572 SEQ ID NO:1340
TGF-beta receptor 2 IL-7Ralpha TM mut31 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:573 SEQ ID NO:1341
TGF-beta receptor 2 IL-7Ralpha TM mut32 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:574 SEQ ID NO:1342
TGF-beta receptor 2 IL-7Ralpha TM mut1 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:575 SEQ ID NO:1343
TGF-beta receptor 2 IL-7Ralpha TM mut2 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:576 SEQ ID NO:1344
TGF-beta receptor 2 IL-7Ralpha TM mut3 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:577 SEQ ID NO:1345
TGF-beta receptor 2 IL-7Ralpha TM mut4 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:578 SEQ ID NO:1346
TGF-beta receptor 2 IL-7Ralpha TM mut5 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:579 SEQ ID NO:1347
TGF-beta receptor 2 IL-7Ralpha TM mut6 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:580 SEQ ID NO:1348
TGF-beta receptor 2 IL-7Ralpha TM mut7 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:581 SEQ ID NO:1349
TGF-beta receptor 2 IL-7Ralpha TM mut8 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:582 SEQ ID NO:1350
TGF-beta receptor 2 IL-7Ralpha TM mut9 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:583 SEQ ID NO:1351
TGF-beta receptor 2 IL-7Ralpha TM mut10 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:584 SEQ ID NO:1352
TGF-beta receptor 2 IL-7Ralpha TM mut11 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:585 SEQ ID NO:1353
TGF-beta receptor 2 IL-7Ralpha TM mut12 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:586 SEQ ID NO:1354
TGF-beta receptor 2 IL-7Ralpha TM mut13 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:587 SEQ ID NO:1355
TGF-beta receptor 2 IL-7Ralpha TM mut14 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:588 SEQ ID NO:1356
TGF-beta receptor 2 IL-7Ralpha TM mut15 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:589 SEQ ID NO:1357
TGF-beta receptor 2 IL-7Ralpha TM mut16 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:590 SEQ ID NO:1358
TGF-beta receptor 2 IL-7Ralpha TM mut17 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:591 SEQ ID NO:1359
TGF-beta receptor 2 IL-7Ralpha TM mut18 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:592 SEQ ID NO:1360
TGF-beta receptor 2 IL-7Ralpha TM mut19 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:593 SEQ ID NO:1361
TGF-beta receptor 2 IL-7Ralpha TM mut20 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:594 SEQ ID NO:1362
TGF-beta receptor 2 IL-7Ralpha TM mut21 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:595 SEQ ID NO:1363
TGF-beta receptor 2 IL-7Ralpha TM mut22 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:596 SEQ ID NO:1364
TGF-beta receptor 2 IL-7Ralpha TM mut23 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:597 SEQ ID NO:1365
TGF-beta receptor 2 IL-7Ralpha TM mut24 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:598 SEQ ID NO:1366
TGF-beta receptor 2 IL-7Ralpha TM mut25 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:599 SEQ ID NO:1367
TGF-beta receptor 2 IL-7Ralpha TM mut26 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:600 SEQ ID NO:1368
TGF-beta receptor 2 IL-7Ralpha TM mut27 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:601 SEQ ID NO:1369
TGF-beta receptor 2 IL-7Ralpha TM mut28 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:602 SEQ ID NO:1370
TGF-beta receptor 2 IL-7Ralpha TM mut29 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:603 SEQ ID NO:1371
TGF-beta receptor 2 IL-7Ralpha TM mut30 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:604 SEQ ID NO:1372
TGF-beta receptor 2 IL-7Ralpha TM mut31 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:605 SEQ ID NO:1373
TGF-beta receptor 2 IL-7Ralpha TM mut32 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:606 SEQ ID NO:1374
TGF-beta receptor 3 IL-7Ralpha TM mut1 Intracellular domain of IL-7Ralpha SEQ ID NO:607 SEQ ID NO:1375
TGF-beta receptor 3 IL-7Ralpha TM mut2 Intracellular domain of IL-7Ralpha SEQ ID NO:608 SEQ ID NO:1376
TGF-beta receptor 3 IL-7Ralpha TM mut3 Intracellular domain of IL-7Ralpha SEQ ID NO:609 SEQ ID NO:1377
TGF-beta receptor 3 IL-7Ralpha TM mut4 Intracellular domain of IL-7Ralpha SEQ ID NO:610 SEQ ID NO:1378
TGF-beta receptor 3 IL-7Ralpha TM mut5 Intracellular domain of IL-7Ralpha SEQ ID NO:611 SEQ ID NO:1379
TGF-beta receptor 3 IL-7Ralpha TM mut6 Intracellular domain of IL-7Ralpha SEQ ID NO:612 SEQ ID NO:1380
TGF-beta receptor 3 IL-7Ralpha TM mut7 Intracellular domain of IL-7Ralpha SEQ ID NO:613 SEQ ID NO:1381
TGF-beta receptor 3 IL-7Ralpha TM mut8 Intracellular domain of IL-7Ralpha SEQ ID NO:614 SEQ ID NO:1382
TGF-beta receptor 3 IL-7Ralpha TM mut9 Intracellular domain of IL-7Ralpha SEQ ID NO:615 SEQ ID NO:1383
TGF-beta receptor 3 IL-7Ralpha TM mut10 Intracellular domain of IL-7Ralpha SEQ ID NO:616 SEQ ID NO:1384
TGF-beta receptor 3 IL-7Ralpha TM mut11 Intracellular domain of IL-7Ralpha SEQ ID NO:617 SEQ ID NO:1385
TGF-beta receptor 3 IL-7Ralpha TM mut12 Intracellular domain of IL-7Ralpha SEQ ID NO:618 SEQ ID NO:1386
TGF-beta receptor 3 IL-7Ralpha TM mut13 Intracellular domain of IL-7Ralpha SEQ ID NO:619 SEQ ID NO:1387
TGF-beta receptor 3 IL-7Ralpha TM mut14 Intracellular domain of IL-7Ralpha SEQ ID NO:620 SEQ ID NO:1388
TGF-beta receptor 3 IL-7Ralpha TM mut15 Intracellular domain of IL-7Ralpha SEQ ID NO:621 SEQ ID NO:1389
TGF-beta receptor 3 IL-7Ralpha TM mut16 Intracellular domain of IL-7Ralpha SEQ ID NO:622 SEQ ID NO:1390
TGF-beta receptor 3 IL-7Ralpha TM mut17 Intracellular domain of IL-7Ralpha SEQ ID NO:623 SEQ ID NO:1391
TGF-beta receptor 3 IL-7Ralpha TM mut18 Intracellular domain of IL-7Ralpha SEQ ID NO:624 SEQ ID NO:1392
TGF-beta receptor 3 IL-7Ralpha TM mut19 Intracellular domain of IL-7Ralpha SEQ ID NO:625 SEQ ID NO:1393
TGF-beta receptor 3 IL-7Ralpha TM mut20 Intracellular domain of IL-7Ralpha SEQ ID NO:626 SEQ ID NO:1394
TGF-beta receptor 3 IL-7Ralpha TM mut21 Intracellular domain of IL-7Ralpha SEQ ID NO:627 SEQ ID NO:1395
TGF-beta receptor 3 IL-7Ralpha TM mut22 Intracellular domain of IL-7Ralpha SEQ ID NO:628 SEQ ID NO:1396
TGF-beta receptor 3 IL-7Ralpha TM mut23 Intracellular domain of IL-7Ralpha SEQ ID NO:629 SEQ ID NO:1397
TGF-beta receptor 3 IL-7Ralpha TM mut24 Intracellular domain of IL-7Ralpha SEQ ID NO:630 SEQ ID NO:1398
TGF-beta receptor 3 IL-7Ralpha TM mut25 Intracellular domain of IL-7Ralpha SEQ ID NO:631 SEQ ID NO:1399
TGF-beta receptor 3 IL-7Ralpha TM mut26 Intracellular domain of IL-7Ralpha SEQ ID NO:632 SEQ ID NO:1400
TGF-beta receptor 3 IL-7Ralpha TM mut27 Intracellular domain of IL-7Ralpha SEQ ID NO:633 SEQ ID NO:1401
TGF-beta receptor 3 IL-7Ralpha TM mut28 Intracellular domain of IL-7Ralpha SEQ ID NO:634 SEQ ID NO:1402
TGF-beta receptor 3 IL-7Ralpha TM mut29 Intracellular domain of IL-7Ralpha SEQ ID NO:635 SEQ ID NO:1403
TGF-beta receptor 3 IL-7Ralpha TM mut30 Intracellular domain of IL-7Ralpha SEQ ID NO:636 SEQ ID NO:1404
TGF-beta receptor 3 IL-7Ralpha TM mut31 Intracellular domain of IL-7Ralpha SEQ ID NO:637 SEQ ID NO:1405
TGF-beta receptor 3 IL-7Ralpha TM mut32 Intracellular domain of IL-7Ralpha SEQ ID NO:638 SEQ ID NO:1406
TGF-beta receptor 3 IL-7Ralpha TM mut1 CD28 intracellular +4-1BB intracellular SEQ ID NO:639 SEQ ID NO:1407
TGF-beta receptor 3 IL-7Ralpha TM mut2 CD28 intracellular +4-1BB intracellular SEQ ID NO:640 SEQ ID NO:1408
TGF-beta receptor 3 IL-7Ralpha TM mut3 CD28 intracellular +4-1BB intracellular SEQ ID NO:641 SEQ ID NO:1409
TGF-beta receptor 3 IL-7Ralpha TM mut4 CD28 intracellular +4-1BB intracellular SEQ ID NO:642 SEQ ID NO:1410
TGF-beta receptor 3 IL-7Ralpha TM mut5 CD28 intracellular +4-1BB intracellular SEQ ID NO:643 SEQ ID NO:1411
TGF-beta receptor 3 IL-7Ralpha TM mut6 CD28 intracellular +4-1BB intracellular SEQ ID NO:644 SEQ ID NO:1412
TGF-beta receptor 3 IL-7Ralpha TM mut7 CD28 intracellular +4-1BB intracellular SEQ ID NO:645 SEQ ID NO:1413
TGF-beta receptor 3 IL-7Ralpha TM mut8 CD28 intracellular +4-1BB intracellular SEQ ID NO:646 SEQ ID NO:1414
TGF-beta receptor 3 IL-7Ralpha TM mut9 CD28 intracellular +4-1BB intracellular SEQ ID NO:647 SEQ ID NO:1415
TGF-beta receptor 3 IL-7Ralpha TM mut10 CD28 intracellular +4-1BB intracellular SEQ ID NO:648 SEQ ID NO:1416
TGF-beta receptor 3 IL-7Ralpha TM mut11 CD28 intracellular +4-1BB intracellular SEQ ID NO:649 SEQ ID NO:1417
TGF-beta receptor 3 IL-7Ralpha TM mut12 CD28 intracellular +4-1BB intracellular SEQ ID NO:650 SEQ ID NO:1418
TGF-beta receptor 3 IL-7Ralpha TM mut13 CD28 intracellular +4-1BB intracellular SEQ ID NO:651 SEQ ID NO:1419
TGF-beta receptor 3 IL-7Ralpha TM mut14 CD28 intracellular domain +4-1BB intracellular region SEQ ID NO:652 SEQ ID NO:1420
TGF-beta receptor 3 IL-7Ralpha TM mut15 CD28 intracellular +4-1BB intracellular SEQ ID NO:653 SEQ ID NO:1421
TGF-beta receptor 3 IL-7Ralpha TM mut16 CD28 intracellular +4-1BB intracellular SEQ ID NO:654 SEQ ID NO:1422
TGF-beta receptor 3 IL-7Ralpha TM mut17 CD28 intracellular +4-1BB intracellular SEQ ID NO:655 SEQ ID NO:1423
TGF-beta receptor 3 IL-7Ralpha TM mut18 CD28 intracellular +4-1BB intracellular SEQ ID NO:656 SEQ ID NO:1424
TGF-beta receptor 3 IL-7Ralpha TM mut19 CD28 intracellular +4-1BB intracellular SEQ ID NO:657 SEQ ID NO:1425
TGF-beta receptor 3 IL-7Ralpha TM mut20 CD28 intracellular +4-1BB intracellular SEQ ID NO:658 SEQ ID NO:1426
TGF-beta receptor 3 IL-7Ralpha TM mut21 CD28 intracellular +4-1BB intracellular SEQ ID NO:659 SEQ ID NO:1427
TGF-beta receptor 3 IL-7Ralpha TM mut22 CD28 intracellular +4-1BB intracellular SEQ ID NO:660 SEQ ID NO:1428
TGF-beta receptor 3 IL-7Ralpha TM mut23 CD28 intracellular +4-1BB intracellular SEQ ID NO:661 SEQ ID NO:1429
TGF-beta receptorBody 3 IL-7Ralpha TM mut24 CD28 intracellular +4-1BB intracellular SEQ ID NO:662 SEQ ID NO:1430
TGF-beta receptor 3 IL-7Ralpha TM mut25 CD28 intracellular +4-1BB intracellular SEQ ID NO:663 SEQ ID NO:1431
TGF-beta receptor 3 IL-7Ralpha TM mut26 CD28 intracellular +4-1BB intracellular SEQ ID NO:664 SEQ ID NO:1432
TGF-beta receptor 3 IL-7Ralpha TM mut27 CD28 intracellular +4-1BB intracellular SEQ ID NO:665 SEQ ID NO:1433
TGF-beta receptor 3 IL-7Ralpha TM mut28 CD28 intracellular +4-1BB intracellular SEQ ID NO:666 SEQ ID NO:1434
TGF-beta receptor 3 IL-7Ralpha TM mut29 CD28 intracellular +4-1BB intracellular SEQ ID NO:667 SEQ ID NO:1435
TGF-beta receptor 3 IL-7Ralpha TM mut30 CD28 intracellular +4-1BB intracellular SEQ ID NO:668 SEQ ID NO:1436
TGF-beta receptor 3 IL-7Ralpha TM mut31 CD28 intracellular +4-1BB intracellular SEQ ID NO:669 SEQ ID NO:1437
TGF-beta receptor 3 IL-7Ralpha TM mut32 CD28 intracellular +4-1BB intracellular SEQ ID NO:670 SEQ ID NO:1438
TGF-beta receptor 3 IL-7Ralpha TM mut1 CD28 intracellular domain SEQ ID NO:671 SEQ ID NO:1439
TGF-beta receptor 3 IL-7Ralpha TM mut2 CD28 intracellular domain SEQ ID NO:672 SEQ ID NO:1440
TGF-beta receptor 3 IL-7Ralpha TM mut3 CD28 intracellular domain SEQ ID NO:673 SEQ ID NO:1441
TGF-beta receptor 3 IL-7Ralpha TM mut4 CD28 intracellular domain SEQ ID NO:674 SEQ ID NO:1442
TGF-beta receptor 3 IL-7Ralpha TM mut5 CD28 intracellular domain SEQ ID NO:675 SEQ ID NO:1443
TGF-beta receptor 3 IL-7Ralpha TM mut6 CD28 intracellular domain SEQ ID NO:676 SEQ ID NO:1444
TGF-beta receptor 3 IL-7Ralpha TM mut7 CD28 intracellular domain SEQ ID NO:677 SEQ ID NO:1445
TGF-beta receptor 3 IL-7Ralpha TM mut8 CD28 intracellular domain SEQ ID NO:678 SEQ ID NO:1446
TGF-beta receptor 3 IL-7Ralpha TM mut9 CD28 intracellular domain SEQ ID NO:679 SEQ ID NO:1447
TGF-beta receptor 3 IL-7Ralpha TM mut10 CD28 intracellular domain SEQ ID NO:680 SEQ ID NO:1448
TGF-beta receptor 3 IL-7Ralpha TM mut11 CD28 intracellular domain SEQ ID NO:681 SEQ ID NO:1449
TGF-beta receptor 3 IL-7Ralpha TM mut12 CD28 intracellular domain SEQ ID NO:682 SEQ ID NO:1450
TGF-beta receptor 3 IL-7Ralpha TM mut13 CD28 intracellular domain SEQ ID NO:683 SEQ ID NO:1451
TGF-beta receptor 3 IL-7Ralpha TM mut14 CD28 intracellular domain SEQ ID NO:684 SEQ ID NO:1452
TGF-beta receptor 3 IL-7Ralpha TM mut15 CD28 intracellular domain SEQ ID NO:685 SEQ ID NO:1453
TGF-beta receptor 3 IL-7Ralpha TM mut16 CD28 intracellular domain SEQ ID NO:686 SEQ ID NO:1454
TGF-beta receptor 3 IL-7Ralpha TM mut17 CD28 intracellular domain SEQ ID NO:687 SEQ ID NO:1455
TGF-beta receptor 3 IL-7Ralpha TM mut18 CD28 intracellular domain SEQ ID NO:688 SEQ ID NO:1456
TGF-beta receptor 3 IL-7Ralpha TM mut19 CD28 intracellular domain SEQ ID NO:689 SEQ ID NO:1457
TGF-beta receptor 3 IL-7Ralpha TM mut20 CD28 intracellular domain SEQ ID NO:690 SEQ ID NO:1458
TGF-beta receptor 3 IL-7Ralpha TM mut21 CD28 intracellular domain SEQ ID NO:691 SEQ ID NO:1459
TGF-beta receptor 3 IL-7Ralpha TM mut22 CD28 intracellular domain SEQ ID NO:692 SEQ ID NO:1460
TGF-beta receptor 3 IL-7Ralpha TM mut23 CD28 intracellular domain SEQ ID NO:693 SEQ ID NO:1461
TGF-beta receptor 3 IL-7Ralpha TM mut24 CD28 intracellular domain SEQ ID NO:694 SEQ ID NO:1462
TGF-beta receptor 3 IL-7Ralpha TM mut25 CD28 intracellular domain SEQ ID NO:695 SEQ ID NO:1463
TGF-beta receptor 3 IL-7Ralpha TM mut26 CD28 intracellular domain SEQ ID NO:696 SEQ ID NO:1464
TGF-beta receptor 3 IL-7Ralpha TM mut27 CD28 intracellular domain SEQ ID NO:697 SEQ ID NO:1465
TGF-beta receptor 3 IL-7Ralpha TM mut28 CD28 intracellular domain SEQ ID NO:698 SEQ ID NO:1466
TGF-beta receptor 3 IL-7Ralpha TM mut29 CD28 intracellular domain SEQ ID NO:699 SEQ ID NO:1467
TGF-beta receptor 3 IL-7Ralpha TM mut30 CD28 intracellular domain SEQ ID NO:700 SEQ ID NO:1468
TGF-beta receptor 3 IL-7Ralpha TM mut31 CD28 intracellular domain SEQ ID NO:701 SEQ ID NO:1469
TGF-beta receptor 3 IL-7Ralpha TM mut32 Intracellular CD28Zone(s) SEQ ID NO:702 SEQ ID NO:1470
TGF-beta receptor 3 IL-7Ralpha TM mut1 CD28 intracellular + OX40 intracellular SEQ ID NO:703 SEQ ID NO:1471
TGF-beta receptor 3 IL-7Ralpha TM mut2 CD28 intracellular + OX40 intracellular SEQ ID NO:704 SEQ ID NO:1472
TGF-beta receptor 3 IL-7Ralpha TM mut3 CD28 intracellular + OX40 intracellular SEQ ID NO:705 SEQ ID NO:1473
TGF-beta receptor 3 IL-7Ralpha TM mut4 CD28 intracellular + OX40 intracellular SEQ ID NO:706 SEQ ID NO:1474
TGF-beta receptor 3 IL-7Ralpha TM mut5 CD28 intracellular + OX40 intracellular SEQ ID NO:707 SEQ ID NO:1475
TGF-beta receptor 3 IL-7Ralpha TM mut6 CD28 intracellular + OX40 intracellular SEQ ID NO:708 SEQ ID NO:1476
TGF-beta receptor 3 IL-7Ralpha TM mut7 CD28 intracellular + OX40 intracellular SEQ ID NO:709 SEQ ID NO:1477
TGF-beta receptor 3 IL-7Ralpha TM mut8 CD28 intracellular + OX40 intracellular SEQ ID NO:710 SEQ ID NO:1478
TGF-beta receptor 3 IL-7Ralpha TM mut9 CD28 intracellular + OX40 intracellular SEQ ID NO:711 SEQ ID NO:1479
TGF-beta receptor 3 IL-7Ralpha TM mut10 CD28 intracellular + OX40 intracellular SEQ ID NO:712 SEQ ID NO:1480
TGF-beta receptor 3 IL-7Ralpha TM mut11 CD28 intracellular + OX40 intracellular SEQ ID NO:713 SEQ ID NO:1481
TGF-beta receptor 3 IL-7Ralpha TM mut12 CD28 intracellular + OX40 intracellular SEQ ID NO:714 SEQ ID NO:1482
TGF-beta receptor 3 IL-7Ralpha TM mut13 CD28 intracellular + OX40 intracellular SEQ ID NO:715 SEQ ID NO:1483
TGF-beta receptor 3 IL-7Ralpha TM mut14 CD28 intracellular + OX40 intracellular SEQ ID NO:716 SEQ ID NO:1484
TGF-beta receptor 3 IL-7Ralpha TM mut15 CD28 intracellular + OX40 intracellular SEQ ID NO:717 SEQ ID NO:1485
TGF-beta receptor 3 IL-7Ralpha TM mut16 CD28 intracellular + OX40 intracellular SEQ ID NO:718 SEQ ID NO:1486
TGF-beta receptor 3 IL-7Ralpha TM mut17 CD28 intracellular + OX40 intracellular SEQ ID NO:719 SEQ ID NO:1487
TGF-beta receptor 3 IL-7Ralpha TM mut18 CD28 intracellular + OX40 intracellular SEQ ID NO:720 SEQ ID NO:1488
TGF-beta receptor 3 IL-7Ralpha TM mut19 CD28 intracellular + OX40 intracellular SEQ ID NO:721 SEQ ID NO:1489
TGF-beta receptor 3 IL-7Ralpha TM mut20 CD28 intracellular + OX40 intracellular SEQ ID NO:722 SEQ ID NO:1490
TGF-beta receptor 3 IL-7Ralpha TM mut21 CD28 intracellular + OX40 intracellular SEQ ID NO:723 SEQ ID NO:1491
TGF-beta receptor 3 IL-7Ralpha TM mut22 CD28 intracellular + OX40 intracellular SEQ ID NO:724 SEQ ID NO:1492
TGF-beta receptor 3 IL-7Ralpha TM mut23 CD28 intracellular + OX40 intracellular SEQ ID NO:725 SEQ ID NO:1493
TGF-beta receptor 3 IL-7Ralpha TM mut24 CD28 intracellular + OX40 intracellular SEQ ID NO:726 SEQ ID NO:1494
TGF-beta receptor 3 IL-7Ralpha TM mut25 CD28 intracellular + OX40 intracellular SEQ ID NO:727 SEQ ID NO:1495
TGF-beta receptor 3 IL-7Ralpha TM mut26 CD28 intracellular + OX40 intracellular SEQ ID NO:728 SEQ ID NO:1496
TGF-beta receptor 3 IL-7Ralpha TM mut27 CD28 intracellular + OX40 intracellular SEQ ID NO:729 SEQ ID NO:1497
TGF-beta receptor 3 IL-7Ralpha TM mut28 CD28 intracellular + OX40 intracellular SEQ ID NO:730 SEQ ID NO:1498
TGF-beta receptor 3 IL-7Ralpha TM mut29 CD28 intracellular + OX40 intracellular SEQ ID NO:731 SEQ ID NO:1499
TGF-beta receptor 3 IL-7Ralpha TM mut30 CD28 intracellular + OX40 intracellular SEQ ID NO:732 SEQ ID NO:1500
TGF-beta receptor 3 IL-7Ralpha TM mut31 CD28 intracellular + OX40 intracellular SEQ ID NO:733 SEQ ID NO:1501
TGF-beta receptor 3 IL-7Ralpha TM mut32 CD28 intracellular + OX40 intracellular SEQ ID NO:734 SEQ ID NO:1502
TGF-beta receptor 3 IL-7Ralpha TM mut1 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:735 SEQ ID NO:1503
TGF-beta receptor 3 IL-7Ralpha TM mut2 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:736 SEQ ID NO:1504
TGF-beta receptor 3 IL-7Ralpha TM mut3 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:737 SEQ ID NO:1505
TGF-beta receptor 3 IL-7Ralpha TM mut4 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:738 SEQ ID NO:1506
TGF-beta receptor 3 IL-7Ralpha TM mut5 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:739 SEQ ID NO:1507
TGF-beta receptor 3 IL-7Ralpha TM mut6 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:740 SEQ ID NO:1508
TGF-beta receptor 3 IL-7Ralpha TM mut7 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:741 SEQ ID NO:1509
TGF-beta receptor 3 IL-7Ralpha TM mut8 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:742 SEQ ID NO:1510
TGF-beta receptor 3 IL-7Ralpha TM mut9 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:743 SEQ ID NO:1511
TGF-beta receptor 3 IL-7Ralpha TM mut10 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:744 SEQ ID NO:1512
TGF-beta receptor 3 IL-7Ralpha TM mut11 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:745 SEQ ID NO:1513
TGF-beta receptor 3 IL-7Ralpha TM mut12 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:746 SEQ ID NO:1514
TGF-beta receptor 3 IL-7Ralpha TM mut13 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:747 SEQ ID NO:1515
TGF-beta receptor 3 IL-7Ralpha TM mut14 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:748 SEQ ID NO:1516
TGF-beta receptor 3 IL-7Ralpha TM mut15 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:749 SEQ ID NO:1517
TGF-beta receptor 3 IL-7Ralpha TM mut16 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:750 SEQ ID NO:1518
TGF-beta receptor 3 IL-7Ralpha TM mut17 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:751 SEQ ID NO:1519
TGF-beta receptor 3 IL-7Ralpha TM mut18 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:752 SEQ ID NO:1520
TGF-beta receptor 3 IL-7Ralpha TM mut19 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:753 SEQ ID NO:1521
TGF-beta receptor 3 IL-7Ralpha TM mut20 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:754 SEQ ID NO:1522
TGF-beta receptor 3 IL-7Ralpha TM mut21 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:755 SEQ ID NO:1523
TGF-beta receptor 3 IL-7Ralpha TM mut22 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:756 SEQ ID NO:1524
TGF-beta receptor 3 IL-7Ralpha TM mut23 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:757 SEQ ID NO:1525
TGF-beta receptor 3 IL-7Ralpha TM mut24 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:758 SEQ ID NO:1526
TGF-beta receptor 3 IL-7Ralpha TM mut25 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:759 SEQ ID NO:1527
TGF-beta receptor 3 IL-7Ralpha TM mut26 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:760 SEQ ID NO:1528
TGF-beta receptor 3 IL-7Ralpha TM mut27 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:761 SEQ ID NO:1529
TGF-beta receptor 3 IL-7Ralpha TM mut28 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:762 SEQ ID NO:1530
TGF-beta receptor 3 IL-7Ralpha TM mut29 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:763 SEQ ID NO:1531
TGF-beta receptor 3 IL-7Ralpha TM mut30 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:764 SEQ ID NO:1532
TGF-beta receptor 3 IL-7Ralpha TM mut31 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:765 SEQ ID NO:1533
TGF-beta receptor 3 IL-7Ralpha TM mut32 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:766 SEQ ID NO:1534
TGF-beta receptor 3 IL-7Ralpha TM mut1 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:767 SEQ ID NO:1535
TGF-beta receptor 3 IL-7Ralpha TM mut2 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:768 SEQ ID NO:1536
TGF-beta receptor 3 IL-7Ralpha TM mut3 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:769 SEQ ID NO:1537
TGF-beta receptor 3 IL-7Ralpha TM mut4 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:770 SEQ ID NO:1538
TGF-beta receptor 3 IL-7Ralpha TM mut5 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:771 SEQ ID NO:1539
TGF-beta receptor 3 IL-7Ralpha TM mut6 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:772 SEQ ID NO:1540
TGF-beta receptor 3 IL-7Ralpha TM mut7 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:773 SEQ ID NO:1541
TGF-beta receptor 3 IL-7Ralpha TM mut8 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:774 SEQ ID NO:1542
TGF-beta receptor 3 IL-7Ralpha TM mut9 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:775 SEQ ID NO:1543
TGF-beta receptor 3 IL-7Ralpha TM mut10 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:776 SEQ ID NO:1544
TGF-beta receptor 3 IL-7Ralpha TM mut11 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:777 SEQ ID NO:1545
TGF-beta receptor 3 IL-7Ralpha TM mut12 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:778 SEQ ID NO:1546
TGF-beta receptor 3 IL-7Ralpha TM mut13 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:779 SEQ ID NO:1547
TGF-beta receptor 3 IL-7Ralpha TM mut14 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:780 SEQ ID NO:1548
TGF-beta receptor 3 IL-7Ralpha TM mut15 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:781 SEQ ID NO:1549
TGF-beta receptor 3 IL-7Ralpha TM mut16 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:782 SEQ ID NO:1550
TGF-beta receptor 3 IL-7Ralpha TM mut17 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:783 SEQ ID NO:1551
TGF-beta receptor 3 IL-7Ralpha TM mut18 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:784 SEQ ID NO:1552
TGF-beta receptor 3 IL-7Ralpha TM mut19 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:785 SEQ ID NO:1553
TGF-beta receptor 3 IL-7Ralpha TM mut20 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:786 SEQ ID NO:1554
TGF-beta receptor 3 IL-7Ralpha TM mut21 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:787 SEQ ID NO:1555
TGF-beta receptor 3 IL-7Ralpha TM mut22 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:788 SEQ ID NO:1556
TGF-beta receptor 3 IL-7Ralpha TM mut23 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:789 SEQ ID NO:1557
TGF-beta receptor 3 IL-7Ralpha TM mut24 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:790 SEQ ID NO:1558
TGF-beta receptor 3 IL-7Ralpha TM mut25 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:791 SEQ ID NO:1559
TGF-beta receptor 3 IL-7Ralpha TM mut26 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:792 SEQ ID NO:1560
TGF-beta receptor 3 IL-7Ralpha TM mut27 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:793 SEQ ID NO:1561
TGF-beta receptor 3 IL-7Ralpha TM mut28 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:794 SEQ ID NO:1562
TGF-beta receptor 3 IL-7Ralpha TM mut29 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:795 SEQ ID NO:1563
TGF-beta receptor 3 IL-7Ralpha TM mut30 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:796 SEQ ID NO:1564
TGF-beta receptor 3 IL-7Ralpha TM mut31 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:797 SEQ ID NO:1565
TGF-beta receptor 3 IL-7Ralpha TM mut32 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:798 SEQ ID NO:1566
TGF-beta receptor 3 IL-7Ralpha TM mut1 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:799 SEQ ID NO:1567
TGF-beta receptor 3 IL-7Ralpha TM mut2 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:800 SEQ ID NO:1568
TGF-beta receptor 3 IL-7Ralpha TM mut3 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:801 SEQ ID NO:1569
TGF-beta receptor 3 IL-7Ralpha TM mut4 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:802 SEQ ID NO:1570
TGF-beta receptor 3 IL-7Ralpha TM mut5 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:803 SEQ ID NO:1571
TGF-beta receptor 3 IL-7Ralpha TM mut6 IL2RB-z (YXXQ) intracellular region SEQ ID NO:804 SEQ ID NO:1572
TGF-beta receptor 3 IL-7Ralpha TM mut7 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:805 SEQ ID NO:1573
TGF-beta receptor 3 IL-7Ralpha TM mut8 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:806 SEQ ID NO:1574
TGF-beta receptor 3 IL-7Ralpha TM mut9 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:807 SEQ ID NO:1575
TGF-beta receptor 3 IL-7Ralpha TM mut10 IL2RB-z (YXXQ) intracellular region SEQ ID NO:808 SEQ ID NO:1576
TGF-beta receptor 3 IL-7Ralpha TM mut11 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:809 SEQ ID NO:1577
TGF-beta receptor 3 IL-7Ralpha TM mut12 IL2RB-z (YXXQ) intracellular region SEQ ID NO:810 SEQ ID NO:1578
TGF-beta receptor 3 IL-7Ralpha TM mut13 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:811 SEQ ID NO:1579
TGF-beta receptor 3 IL-7Ralpha TM mut14 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:812 SEQ ID NO:1580
TGF-beta receptor 3 IL-7Ralpha TM mut15 IL2RB-z (YXXQ) intracellular region SEQ ID NO:813 SEQ ID NO:1581
TGF-beta receptor 3 IL-7Ralpha TM mut16 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:814 SEQ ID NO:1582
TGF-beta receptor 3 IL-7Ralpha TM mut17 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:815 SEQ ID NO:1583
TGF-beta receptor 3 IL-7Ralpha TM mut18 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:816 SEQ ID NO:1584
TGF-beta receptor 3 IL-7Ralpha TM mut19 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:817 SEQ ID NO:1585
TGF-beta receptor 3 IL-7Ralpha TM mut20 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:818 SEQ ID NO:1586
TGF-beta receptor 3 IL-7Ralpha TM mut21 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:819 SEQ ID NO:1587
TGF-beta receptor 3 IL-7Ralpha TM mut22 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:820 SEQ ID NO:1588
TGF-beta receptor 3 IL-7Ralpha TM mut23 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:821 SEQ ID NO:1589
TGF-beta receptor 3 IL-7Ralpha TM mut24 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:822 SEQ ID NO:1590
TGF-beta receptor 3 IL-7Ralpha TM mut25 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:823 SEQ ID NO:1591
TGF-beta receptor 3 IL-7Ralpha TM mut26 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:824 SEQ ID NO:1592
TGF-beta receptor 3 IL-7Ralpha TM mut27 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:825 SEQ ID NO:1593
TGF-beta receptor 3 IL-7Ralpha TM mut28 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:826 SEQ ID NO:1594
TGF-beta receptor 3 IL-7Ralpha TM mut29 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:827 SEQ ID NO:1595
TGF-beta receptor 3 IL-7Ralpha TM mut30 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:828 SEQ ID NO:1596
TGF-beta receptor 3 IL-7Ralpha TM mut31 IL2RB-z (YXXQ) intracellular region SEQ ID NO:829 SEQ ID NO:1597
TGF-beta receptor 3 IL-7Ralpha TM mut32 Intracellular region of IL2RB-z (YXXQ) SEQ ID NO:830 SEQ ID NO:1598
TGF-beta receptor 3 IL-7Ralpha TM mut1 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:831 SEQ ID NO:1599
TGF-beta receptor 3 IL-7Ralpha TM mut2 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:832 SEQ ID NO:1600
TGF-beta receptor 3 IL-7Ralpha TM mut3 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:833 SEQ ID NO:1601
TGF-beta receptor 3 IL-7Ralpha TM mut4 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:834 SEQ ID NO:1602
TGF-beta receptor 3 IL-7Ralpha TM mut5 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:835 SEQ ID NO:1603
TGF-beta receptor 3 IL-7Ralpha TM mut6 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:836 SEQ ID NO:1604
TGF-beta receptor 3 IL-7Ralpha TM mut7 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:837 SEQ ID NO:1605
TGF-beta receptor 3 IL-7Ralpha TM mut8 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:838 SEQ ID NO:1606
TGF-beta receptor 3 IL-7Ralpha TM mut9 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:839 SEQ ID NO:1607
TGF-beta receptor 3 IL-7Ralpha TM mut10 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:840 SEQ ID NO:1608
TGF-beta receptor 3 IL-7Ralpha TM mut11 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:841 SEQ ID NO:1609
TGF-beta receptor 3 IL-7Ralpha TM mut12 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:842 SEQ ID NO:1610
TGF-beta receptor 3 IL-7Ralpha TM mut13 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:843 SEQ ID NO:1611
TGF-beta receptor 3 IL-7Ralpha TM mut14 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:844 SEQ ID NO:1612
TGF-beta receptor 3 IL-7Ralpha TM mut15 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:845 SEQ ID NO:1613
TGF-beta receptor 3 IL-7Ralpha TM mut16 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:846 SEQ ID NO:1614
TGF-beta receptor 3 IL-7Ralpha TM mut17 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:847 SEQ ID NO:1615
TGF-beta receptor 3 IL-7Ralpha TM mut18 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:848 SEQ ID NO:1616
TGF-beta receptor 3 IL-7Ralpha TM mut19 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:849 SEQ ID NO:1617
TGF-beta receptor 3 IL-7Ralpha TM mut20 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:850 SEQ ID NO:1618
TGF-beta receptor 3 IL-7Ralpha TM mut21 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:851 SEQ ID NO:1619
TGF-beta receptor 3 IL-7Ralpha TM mut22 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:852 SEQ ID NO:1620
TGF-beta receptor 3 IL-7Ralpha TM mut23 DeltaIL2RB-z (YXXQ) intracellularlyZone(s) SEQ ID NO:853 SEQ ID NO:1621
TGF-beta receptor 3 IL-7Ralpha TM mut24 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:854 SEQ ID NO:1622
TGF-beta receptor 3 IL-7Ralpha TM mut25 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:855 SEQ ID NO:1623
TGF-beta receptor 3 IL-7Ralpha TM mut26 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:856 SEQ ID NO:1624
TGF-beta receptor 3 IL-7Ralpha TM mut27 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:857 SEQ ID NO:1625
TGF-beta receptor 3 IL-7Ralpha TM mut28 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:858 SEQ ID NO:1626
TGF-beta receptor 3 IL-7Ralpha TM mut29 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:859 SEQ ID NO:1627
TGF-beta receptor 3 IL-7Ralpha TM mut30 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:860 SEQ ID NO:1628
TGF-beta receptor 3 IL-7Ralpha TM mut31 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:861 SEQ ID NO:1629
TGF-beta receptor 3 IL-7Ralpha TM mut32 DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:862 SEQ ID NO:1630
Also included in the invention are signal transducing receptors comprising a CD28 transmembrane region and a 28-DeltaIL2RB-z (YXXQ) intracellular region or a 28-IL2RB-z (YXXQ) intracellular region, exemplary signal transducing receptors include, but are not limited to, the signal transducing receptors shown in Table 5:
TABLE 5
Extracellular region Transmembrane region Intracellular region Sequence number (protein) Sequence No. (nucleic acid)
TGF-beta receptor 1 CD28 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:1633 SEQ ID NO:1639
TGF-beta receptor 1 CD28 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:1634 SEQ ID NO:1640
TGF-beta receptor 2 CD28 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:1635 SEQ ID NO:1641
TGF-beta receptor 2 CD28 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:1636 SEQ ID NO:1642
TGF-beta receptor 3 CD28 28-DeltaIL2RB-z (YXXQ) intracellular region SEQ ID NO:1637 SEQ ID NO:1643
TGF-beta receptor 3 CD28 28-IL2RB-z (YXXQ) intracellular region SEQ ID NO:1638 SEQ ID NO:1644
IL-7Ralpha TM mut1-32 is disclosed in Shocha C et al, "Gain-of-function criteria in interfacial mechanism-7 receptor-alpha (IL 7R) in children's acid lysine legal letters, J Exp Med.2011, 5/9, 208 (5): 901-8, the entire contents of which are incorporated herein by reference.
28-IL2RB-z (YXXQ), 28-DeltaIL2RB-z (YXXQ), 28-IL2RB-z (YXXQ) intracellular domains and 28-DeltaIL2RB-z (YXXQ) intracellular domains are disclosed in Yuki Kagoya et al, "A Novel Chimeric antibody Receptor containment a JAK-STAT signalling Domain media Superior antibody Effects", nat Med, 3.2018, 24 (3): 352-359, which are herein incorporated by reference in their entirety, with exemplary amino acid sequences and corresponding coding sequences as shown in SEQ ID NOS: 13 and 14, SEQ ID NOS: 15 and 16, SEQ ID NOS: 17 and 18, and SEQ ID NOS: 19 and 20, respectively. IL2RB-z (YXXQ) intracellular regions (exemplary amino acid sequence and coding sequence are shown in SEQ ID NOS: 21 and 22, respectively) and DeltaIL2RB-z (YXXQ) intracellular regions (exemplary amino acid sequence and coding sequence are shown in SEQ ID NOS: 23 and 24, respectively) are 28-IL2RB-z (YXXQ) intracellular regions and 28-DeltaIL2RB-z (YXXQ) intracellular regions, respectively, do not contain portions of the intracellular portion of CD 28.
Preferably, the signal transduction receptor further comprises a signal peptide. Preferably, the signal peptide is located at the N-terminus of the signal transduction receptor. The signal peptide may be any signal peptide conventional in the art capable of directing the polypeptide out of the nucleus, including but not limited to CD8, CD4, CD28, CD137, EGFR, TGFBRI, TGFBRII, TGFBRIII, and light chain signal peptides.
It will be appreciated that, if desired, the extracellular domain and transmembrane region, and/or transmembrane region and intracellular domain described herein may be linked by a linker sequence. Linker sequences well known in the art, such as G and S containing linker sequences, e.g. (GSSS) n Or (GSSSS) n Wherein n is an integer of 1 to 8.
In some embodiments, the signal transducing receptors of the present invention may further comprise a signal peptide sequence. Typically, the signal peptide sequence is linked to the extracellular domain. Exemplary signal peptide sequences include the signal peptide of the TGF-beta receptor. In some embodiments, the TGF- β receptor signal peptide is amino acids 1-22 of TGF- β receptor 2 (SEQ ID NO: 1631), and the coding sequence is shown in SEQ ID NO: 1632.
In some embodiments, the signal transduction receptor of the present invention may further comprise a single-chain antibody or native peptide fragment that recognizes and binds to a tumor-specific antigen or a tumor-associated antigen, such that immune activation is further enhanced upon contacting a T cell expressing the molecular converter with a tumor cell. The tumor specific or tumor associated antigens include CD19, CD20, CEA, GD2, FR, PSMA, gp100, CA9, CD171/L1-CAM, IL-13R α 2, MART-1, ERBB2, NY-ESO-1, MAGE family protein, BAGE family protein, GAGE family protein, AFP, MUC1, CD22, CD23, CD30, CD33, CD44v7/8, CD70, VEGFR1, VEGFR2, IL-11R α, EGP-2, EGP-40, FBP, GD3, PSCA, FSA, PSA, TAG-72, h5T4, fetal acetylcholine receptor, leY, epCAM, MSLN, IGFR1, EGFRvIII, ERBB3, AFERBB 4 CA, AFCA 15-3, GF 19-9 CA72-4, CA242, CA50, CY 21-1, POSCC, FRAV, TSFVCA, TSAV, GRV-2, GRSF 2, and PROSF 2.
Chimeric Antigen Receptor (CAR)
The immune cells of the invention can further express a CAR, or contain a coding sequence for a CAR. The CAR described herein can be any CAR known in the art.
The CAR may comprise, in order, a polypeptide that binds to a tumor cell membrane antigen (e.g., scFv), a hinge region, a transmembrane region, and an intracellular signal region. The CAR of the invention may be constructed using hinge, transmembrane and intracellular signal regions well known in the art for the construction of CARs. In general, a polypeptide that binds to a tumor cell membrane antigen, which is capable of binding with moderate affinity to a membrane antigen that is widely expressed by tumor cells, is typically inserted with an antigenic epitope at any 1,2, or 3 of the following 3 positions: the N-terminus of the polypeptide, between the polypeptide and the hinge region, and within the polypeptide. The polypeptide combined with the tumor cell membrane antigen is natural polypeptide or artificially synthesized polypeptide; preferably, the artificially synthesized polypeptide is a single chain antibody or a Fab fragment.
The chimeric antigen receptor of the present invention may be directed against one or more of the following antigens: CD19, CD20, CEA, GD2 (also known as B4GALNT1, β β GAL acetyl-galactosyltransferase 1), FR (Flavin reductase), PSMA (prostate specific membrane antigen), PMEL pre-melanosome protein), CA9 (carbonic anhydrase IX), CD171/L1-CAM, IL-13RL1, MART-1 (also known as mucin-A), ERBB2, NY-ESO-1 (also known as CTAG1B, cancer/testis antigen 1B), MAGE (melanoma associated antigen E1) family protein, BAGE (B melanoma antigen family) family protein, GAGE (growth hormone releasing factor) family protein, AFP (α fetoprotein), MUC1 (mucin 1, cell surface associated), CD22, CD23, CD30, CD33, CD44v7/8, CD70, VEGFR1, VEGFR2, IL-11R/, EGP-2, EGP-40, FBP, FBST 8 (also known as GD 1A 1), ST8SIA1 acetyl-ceramide alpha Via amide alpha A sialic acid transferase 1), PSCA (prostate Stem cell antigen), FSA (also known as KIAA 1109), PSA (also known as KLK3, kallikrein associated peptidase 3), HMGA2, fetal acetylcholine receptor, leY (also known as FUT 3), epCAM, MSLN (mesothelin), IGFR1, EGFR, EGFRvIII, ERBB3, ERBB4, CA125 (also known as MUC16, mucin 16, cell surface associated), CA15-3, CA19-9, CA72-4, CA242, CA50, CYFRA21-1, SCC (also known as SERPINB 3), AFU (also known as FUCA 1), EBV-VCA, POA (also known as VDR, vitamin D (8978 zx8978-dihydrovitamin D3) receptor, beta body), G (beta), microglobulin), and PROGRP (GRP gastrin-releasing peptide).
A single cell may express multiple CARs, including CARs targeted to different tumor antigens.
T Cell Receptor (TCR) -T
The immune cells of the invention may further express exogenous TCR or contain a coding sequence that expresses an exogenous TCR gene. The TCRs of the invention may be any of a variety of TCRs known in the art, for example, those that are HLA-typing compatible, of known sequence and structure, and that bind antigenic peptide sequences that are also known.
The exogenous TCRs described herein comprise α β double chains that can form a complete TCR complex with double chain structures of γ ∈, δ ∈, and ξ ξ expressed endogenously by immune effector cells such as T cells. The exogenous gene encoding exogenous TCR of the invention comprises an alpha beta double-chain gene, wherein alpha chain and beta chain coding sequences are covalently linked through a linker sequence which can be cut off in vivo, such as a coding DNA sequence of a P2A, T A or F2A sequence, and can also be covalently linked through a DNA fragment encoding an IRES sequence. In addition to the α β double strand encoding the exogenous TCR, the gene encoding the exogenous TCR of the present invention may further comprise a tag protein gene, such as EGFP, RFP, YFP gene, etc., which is expressed by fusion with the α β gene. The tag protein gene may be covalently linked to the α β double stranded gene by a linker sequence that can be cleaved in vivo, such as a 2A sequence, e.g., a DNA sequence encoding P2A, T a or F2A, or by a DNA sequence encoding an IRES sequence. The tag protein, such as EGFP, RFP, YFP gene and the like, is expressed together with the TCR alpha beta double chain and can be used as an identification index for detecting exogenous TCR expression.
The TCR-ts of the invention may be directed against one or more of the following antigens: CD19, CD20, CEA, GD2 (also known as B4GALNT1, β β GAL acetyl-aminogalactosyltransferase 1), FR (Flavin reductase), PSMA (prostate specific membrane antigen), PMEL pre-melanosome protein), CA9 (carbonic anhydrase IX), CD171/L1-CAM, IL-13RL1, MART-1 (also known as mucin-A), ERBB2, NY-ESO-1 (also known as CTAG1B, cancer/testis antigen 1B), MAGE (melanoma associated antigen E1) family protein, BAGE (B melanoma antigen family) family protein, GAGE (growth hormone releasing factor) family protein, AFP (α -fetoprotein), MUC1 (mucin 1, cell surface associated), CD22, CD23, CD30, CD33, CD44v7/8, CD70, VEGFR1, VEGFR2, EGIL-11R/, EGP-2, EGP-40, FBP, FBST 3 (also known as GD 1A 1), ST8SIA1 acetyl-ceramide alpha Via amide alpha A sialic acid transferase 1), PSCA (prostate stem cell antigen), FSA (also known as KIAA 1109), PSA (also known as KLK3, kallikrein-related peptidase 3), HMGA2, fetal acetylcholine receptor, leY (also known as FUT 3), epCAM, MSLN (mesothelin), IGFR1, EGFR, EGFRvIII, ERBB3, ERBB4, CA125 (also known as MUC16, mucin 16, cell surface-related), CA15-3, CA19-9, CA72-4, CA242, CA50, CYFRA21-1, SCC (also known as SERPINB 3), AFU (also known as FUCA 1), EBV-VCA, POA (also known as VDR, vitamin D (1,25-dihydrovitamin D3) receptor), and, betasomes), G (β), microglobulin), and PROGRP (GRP gastrin releasing peptide).
A single cell can express multiple exogenous TCRs, including exogenous TCRs that target different tumor antigens.
Polynucleotide molecules
The present invention provides polynucleotide molecules encoding the signal transducing receptors described herein. The invention also provides the complementary sequence of the coding sequence of the signal transduction receptor. The polynucleotide molecule may be a recombinant nucleic acid molecule or may be synthetic; it may comprise DNA, RNA and PNA (peptide nucleic acid) and may be a hybrid thereof.
Also provided is an expression cassette for a signal transduction receptor of the present invention, which is a nucleic acid construct comprising a promoter, a signal transduction receptor coding sequence, and a PolyA-tailed signal sequence. Other elements required for expression may also be included in the nucleic acid construct, including but not limited to enhancers and the like.
Also provided is a vector comprising a polynucleotide molecule, expression cassette or nucleic acid construct as described herein. The vector may be a plasmid, a cosmid, a virus, or a phage. The vector may be a cloning vector or an expression vector. The expression vector may be a transposon vector. In certain embodiments, the expression vector is one or more of the following transposon vectors: piggybac, sleeping beauty, frog prince, tn5, and Ty. In addition to the polynucleotide molecules of the invention, expression vectors typically contain other elements that are typically included in the vector, such as multiple cloning sites, resistance genes, replication initiation sites, and the like. In certain embodiments, the recombinant expression vector employs as a backbone a pUC18, pUC19, pMD18-T, pMD-T, pGM-T vector, pUC57, pMAX, or pDC315 series vector. In other embodiments, the recombinant expression vector uses a pCDNA3 series vector, a pCDNA4 series vector, a pCDNA5 series vector, a pCDNA6 series vector, a pRL series vector, a pUC57 vector, a pMAX vector, or a pDC315 series vector as a backbone. In certain embodiments, the present invention uses a pSN vector constructed in CN 201510638974.7.
The CAR of the invention may also be expressed in an immune cell according to the invention by a conventional vector. The vector may be a conventional vector for expressing the CAR, including, but not limited to, the various transposon vectors and recombinant expression vectors described above.
In some embodiments, the same vector encodes both the signal converting receptor of the invention and the CAR. The vector may be a dicistronic vector. The coding sequence for the CAR can be positioned 5 'or 3' to the signaling receptor coding sequence. Expression of the CAR and the signaling receptor may be under the direction of the same or different regulatory sequences.
In the case of known polynucleotide sequences, the polynucleotide molecules can be prepared and the corresponding vectors constructed using methods conventional in the art. Recombinant vectors can be constructed using methods well known to those skilled in the art, see, e.g., techniques described in Sambrook et al, ausubel (1989), or other standard texts. Alternatively, the nucleic acid molecule and vector can be reconstituted into liposomes for delivery to target cells. The vectors containing the nucleic acid molecules of the invention can be transferred into host cells by well-known methods, which vary depending on the type of cellular host. For example, calcium chloride transfection is commonly used for prokaryotic cells, while calcium phosphate treatment or electroporation may be used for other cellular hosts, see Sambrook.
Host cell
Herein, when expressing a heterologous nucleic acid sequence, a "host cell" refers to a eukaryotic cell that is capable of replicating a vector and/or expressing a heterologous gene encoded by the vector. The host cell may serve as an acceptor for the vector. A host cell may be "transfected" or "transformed," which refers to the process of transfection or transduction of an exogenous nucleic acid into a host cell. Transformed cells include primary subject cells and their progeny. The terms "engineered" and "recombinant" cells or host cells as used herein often refer to cells into which an exogenous nucleic acid sequence, such as a vector, has been introduced. Thus, a recombinant cell can be distinguished from a naturally occurring cell that does not contain an introduced recombinant nucleic acid.
Herein, host cells include cells that carry the polynucleotide molecules and/or polypeptides described herein. In particular, the invention provides cells carrying the signal transduction receptors of the invention and/or their coding sequences. The cells of the invention are preferably immune cells and can be used for adoptive cell therapy of tumors. Such cells of the invention are also referred to as signal-transducing receptor modified cells of the invention.
More specifically, the cells of the invention are preferably immune effector cells, including T cells, such as cytotoxic T cells (also known as TC, cytotoxic T lymphocytes, CTL, T killer cells, cytolytic T cells, CD8+ T cells, or killer T cells); an NK cell; NKT cells; and other immune cells that can elicit effector functions.
Herein, the cells may be autologous cells, syngeneic cells, allogeneic cells, and even in some cases, xenograft cells, relative to the individual receiving them.
The nucleic acid constructs/recombinant expression vectors of the invention can be transferred into a cell of interest. The method for transferring into the gene is conventional in the art, and includes but is not limited to: viral transduction, microinjection, particle bombardment, biolistic transformation, electrotransformation, and the like. In certain embodiments, the nucleic acid construct or recombinant expression vector is electroporated.
In addition to carrying a signaling receptor and/or coding sequence thereof as described herein, a cell of the invention may also have one or more additional properties useful in cellular immunotherapy (e.g., adoptive cell therapy of tumors). Such other properties may be inherent to the cell or may be part of the cell after genetic manipulation by a human. For example, the cells of the invention may carry chimeric antigen receptors, α β T cell receptors, and/or antigen-specific receptors, such as tumor-specific receptors, or coding sequences thereof.
Pharmaceutical composition
Herein, "pharmaceutical composition" refers to a composition for administration to an individual and encompasses a composition of cells for immunotherapy. The pharmaceutical compositions of the present invention may also comprise a pharmaceutically acceptable carrier. Examples of suitable pharmaceutical carriers are known in the art and include phosphate buffered saline solutions, water, emulsions, such as oil/water emulsions, various types of wetting agents, sterile solutions, and the like. Compositions comprising such carriers can be formulated by well-known conventional methods. These pharmaceutical compositions may be administered to a subject at a suitable dosage.
The dosage regimen may be determined by the attending physician and clinical factors. As is well known in the medical arts, the dosage for any one patient depends on a variety of factors including the patient's size, body surface area, age, the particular compound to be administered, sex, time and route of administration, general health, and other drugs being administered concurrently.
The compositions of the present invention may be administered locally or systemically. In certain embodiments, the compositions provided herein (e.g., cells expressing a signal transducing receptor according to the present invention) may be administered parenterally, e.g., intravenously, intraarterially, intrathecally, subdermally, or intramuscularly. In certain other embodiments, DNA encoding a construct provided herein can be administered directly to a target site, e.g., delivered to an internal or external target site by a gene gun or to an intra-arterial site by a catheter. In a preferred embodiment, the pharmaceutical composition is administered subcutaneously, and in a more preferred embodiment, intravenously. Parenteral formulations include sterile aqueous or nonaqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, ringer's dextrose, dextrose and sodium chloride, lactated ringer's solution, or fixed oils. Intravenous carriers include liquid and nutritional supplements, electrolyte supplements (such as those based on ringer's dextrose), and the like. Preservatives and other additives may also be present, such as, for example, antimicrobials, antioxidants, chelating agents, and inert gases and the like. Furthermore, the pharmaceutical composition of the invention may comprise a proteinaceous carrier, such as serum albumin or immunoglobulin, preferably of human origin. In addition to the proteinaceous chimeric cytokine receptor construct or the nucleic acid molecule or vector encoding it, it is contemplated that the pharmaceutical composition of the present invention may further comprise a biologically active agent, depending on the intended use of the pharmaceutical composition.
Method and use
The signal conversion receptor, the polynucleotide molecule, the vector, the host cell and the pharmaceutical composition containing the substances can be used for preventing, treating or relieving cancers, particularly cancers with corresponding tumor antigens expressed on the surfaces of cancer cells, or used for preparing medicaments for preventing, treating or relieving the cancers.
As used herein, "treating" or "treatment" includes any beneficial or desired effect on the symptoms or pathology of a disease or pathological condition, and may include even a small reduction in one or more measurable markers of the disease or disorder (e.g., cancer) under treatment. Treatment may optionally include reduction or alleviation of the symptoms of the disease or disorder, or delay in the progression of the disease or disorder. "treatment" does not necessarily mean complete eradication or cure of the disease or disorder or symptoms associated therewith.
As used herein, "prevention" refers to a method for preventing, inhibiting, or reducing the likelihood of occurrence or recurrence of a disease or disorder (e.g., cancer). It also refers to delaying the onset or recurrence of a disease or disorder or delaying the onset or recurrence of symptoms of a disease or disorder. As used herein, "preventing" also includes reducing the intensity, impact, symptoms, and/or burden of a disease or disorder before it occurs or recurs.
The invention encompasses the administration of cells, polynucleotide molecules and vectors, alone or in any combination, optionally together with pharmaceutically acceptable carriers or excipients, using standard vectors and/or gene delivery systems. In certain embodiments, the polynucleotide molecule or vector may be stably integrated into the genome of the subject following administration.
In particular embodiments, viral vectors that are specific for and persist in certain cells or tissues may be used. Suitable pharmaceutical carriers and excipients are well known in the art. The compositions prepared according to the invention may be used for the prevention or treatment or delay of progression of the above identified diseases.
Further, the present invention provides a method for preventing, treating or ameliorating cancer, comprising the steps of: administering to a subject in need thereof an effective amount of cells carrying a signal-transducing receptor, polynucleotide molecule and/or vector as described herein and/or generated by a method described herein.
Herein, cancers include, but are not limited to, breast, prostate, lung and colon or epithelial cancers, such as breast, colon, prostate, head and neck, skin cancers; genito-urinary tract cancers, such as ovarian cancer, endometrial cancer, cervical cancer; kidney cancer, lung cancer, stomach cancer, small intestine cancer, liver cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, esophageal cancer, salivary gland cancer, thyroid cancer, etc. Administration of the compositions of the invention can be used at all stages and types of cancer, including for example, minimal residual disease, early stage cancer, late stage cancer, and/or metastatic cancer and/or cancer that is refractory to treatment.
By way of example, a cancer patient or a patient susceptible to cancer or a patient suspected of having cancer is treated as follows. Cells modified as described herein can be administered to an individual and left for an extended period of time. The subject may receive one or more administrations of the cells, and the interval between administrations may be days, weeks, months or years. In particular embodiments, multiple administrations may occur over several weeks or months, e.g., 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more weeks or months. In some embodiments, the genetically modified cells are encapsulated to inhibit immune recognition and are located at a tumor site. In the case where cells are provided to an individual after a tumor has recurred after initial treatment with cells of the invention, these cells can be altered to recognize different target tumor antigens. For example, where an initial round includes cells carrying a signaling receptor of the invention and another receptor specific for a particular antigen, receptors for different particular antigens may be used after subsequent rounds, including after tumor recurrence.
In some embodiments, an effective amount of a therapeutic cell that carries or expresses a signaling receptor according to any embodiment of the invention and optionally a CAR or exogenous transgenic TCR is provided to an individual in need thereof. These cells may be delivered at the same time or at a different time as one or more other cancer treatments. These cells and other cancer therapeutics can be delivered in the same or separate formulations. The cells and other cancer therapeutic agents can be provided to the subject in separate delivery routes. Cells and/or other cancer therapeutics can be delivered by, for example, injection at the tumor site or intravenous or oral administration. Conventional routes of delivery for such compositions are known in the art.
The number of cells employed will depend on a variety of circumstances, such as the purpose of introduction, the lifespan of the cells, the protocol to be used, the number of administrations, the capacity of the cells to multiply, the stability of the recombinant construct, and the like.
Cells may be administered as desired. In some embodiments, multiple recipes may be used to adjust recipe parameters. In particular embodiments, the route or number or timing of administration, the lifespan of the cells, and/or the number of cells present may be varied. The number of administrations may depend, for example, at least in part on the factors described above.
Reagent kit
Any of the compositions described herein can be included in a kit. In one non-limiting example, cells expressing a signal transduction receptor according to any one of the embodiments of the present invention and/or reagents for generating one or more cells for cell therapy comprising a recombinant expression vector may be included in a kit. The kit components are provided in suitable containers.
Some of the components of these kits may be packaged in an aqueous matrix or in lyophilized form. The container means of these kits generally comprise at least one vial, test tube, flask, bottle, syringe or other container means in which the components may be placed and preferably suitably dispensed. Where more than one component is present in a kit, the kit will typically also contain a second, third or other container in which the other components can be separately placed. However, various combinations of components may be included in the vial. The kits of the invention will also typically comprise means for containing the component in a commercially available, closed confinement form. Such containers may include injection or blow molded plastic containers in which the desired vials are retained.
When the components of the kit are provided in one and/or more liquid solutions, the liquid solution is an aqueous solution, with a sterile aqueous solution being particularly preferred. In some cases, the receptacle means itself may be a syringe, pipette, and/or other such device.
The components of the kit may also be provided in dry powder form. When the reagents and/or components are provided as dry powders, the powders may be reconstituted by the addition of a suitable solvent. Thus, the kit may further comprise a second container means comprising a sterile, pharmaceutically acceptable buffer and/or other diluent.
In a specific embodiment of the invention, the cells to be used in the cell therapy described herein are provided in a kit. In some embodiments, the cell is substantially the only component of the kit. The kit may contain reagents and materials for preparing the desired cells. In particular embodiments, the reagents and materials comprise primers, nucleotides, suitable buffers or buffering reagents, salts, and the like for amplifying a desired sequence, and in some cases, the reagents comprise DNA and/or vectors encoding a signal transduction receptor and/or a regulatory element thereof as described in any embodiment herein.
Effects of the invention
The invention creatively connects the polypeptide, the transmembrane domain peptide segment and the co-stimulation signal molecule intracellular peptide segment which are efficiently combined with the tumor cell surface and/or tumor stroma cell surface immunosuppressive molecules to form the fusion protein with the function of a T signal molecule converter, and the T cell modified by the fusion protein has the immunosuppressive effect after contacting the tumor cell surface and/or tumor stroma cell surface, is not negatively influenced by the immunosuppressive molecules, but has the immune activation effect, enhances the proliferation capacity of the T cell and the secretion function of cytokines, prolongs the survival time of the activated immune cell, overcomes the side effect of a tumor immunosuppressive microenvironment on tumor adoptive cell treatment effector cells, and effectively improves the tumor inhibition and killing effects of the immune effector cells in vivo and in vitro.
Embodiments of the present invention will be described in detail with reference to examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples do not show the specific techniques or conditions, and the techniques or conditions are described in the literature in the art (for example, refer to J. SammBruk et al, molecular cloning, A laboratory Manual, third edition, science Press, translated by Huangpetang et al), the corresponding references, or the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
EXAMPLE 1 construction of Signal transduction receptor expression vectors
The pNB vector was constructed according to the method described in example 1, page 15 of the specification of Chinese patent CN 105154473B. A pNB vector containing an expression cassette for the TGF- β signaling receptor 2-1-TGF- β signaling receptor 2-15 (TGFBR 2-SCR1-TGFBR2-SCR 15) was constructed according to the method described in CN105154473B, page 15, example 2.
Chinese patent application CN105452287a describes a signal transduction receptor comprising the extracellular domain of TGF- β receptor 2, and example 1 on page 21 of the specification describes the structures of two signal transduction receptors, RIID2 and RIID4, comprising TGF- β signal transduction receptor 2 signal peptide, TGF- β signal transduction receptor 2 extracellular domain, TLR2 transmembrane domain, TLR2 intracellular domain signal transduction receptor and TGF- β signal transduction receptor 2 signal peptide, TGF- β signal transduction receptor 2 extracellular domain, TLR4 transmembrane domain, TLR4 intracellular domain, respectively. According to the above-described structure, signal-converting receptors SCR-RIID2 and SCR-RIID4 including the above-described element were synthesized, and neither contained IRES and either orange fluorescent protein (mOrange) or Green Fluorescent Protein (GFP).
The structures corresponding to TGFBR2-SCR1-TGFBR2-SCR15, SCR-RIID2 and SCR-RIID4 are shown in the following table 6:
TABLE 6
Figure PCTCN2021097361-APPB-000004
Figure PCTCN2021097361-APPB-000005
Specifically, the company was entrusted to synthesize SEQ ID NO:1648, SEQ ID NO:1650, SEQ ID NO:1652, SEQ ID NO:1654, SEQ ID NO:1656, SEQ ID NO:1658, SEQ ID NO:1660, SEQ ID NO:1662, SEQ ID NO:1664, SEQ ID NO:1666, SEQ ID NO:1668, SEQ ID NO:1670, SEQ ID NO:1672, SEQ ID NO:1674, SEQ ID NO:1676, SEQ ID NO:1678, and SEQ ID NO:1680, wherein the sequences are cloned into prepared pNB vectors according to the method described in example 2 at page 15 of the CN105154473B description and named pNB328-1648, pNB328-1650, pNB328-1652, pNB328-1654, pNB328-1656, pNB328-1658, pNB328-1660, pNB328-1662, pNB328-1664, pNB 328-328, pNB328-1658, pNB328-1660, pNB328-1668, pNB328-1670, pNB328-1668, pNB328-1662, pNB328-1668, pNB 328-328, pNB-328-1670 and pNB-328-1670.
Coli (DH 5 c), and after sequencing was correct, the plasmids were extracted and purified using a plasmid purification kit from Qiagen, to obtain high-quality plasmids of each recombinant expression vector.
Example 2 preparation of NY-ESO-1 TCR expression vectors
Synthesizing the gene DNA sequences of alpha chain and beta chain of TCR for recognizing NY-ESO-1 antigen peptide SLLMWITQC (HLA-02). And then the 3' end of SEQ ID NO. 1681 is connected with a DNA sequence for coding EGFP by a DNA for coding a P2A peptide segment, and the obtained sequence is shown as SEQ ID NO. 1682. The company was entrusted with the synthesis of SEQ ID NO:1682, and the clone of SEQ ID NO:1682 into the prepared pNB vector was designated pNB328-1682 according to the method described in example 2 at page 15 of the CN105154473B instructions. And deleting a PB transposase coding region and a CMV promoter in the pNB vector to obtain a vector pNC without a PB transposase expression frame. The clone of SEQ ID NO 1682 into the prepared pNC vector was designated pNC328-1682 according to the method described in CN105154473B, page 15, example 2. Coli (DH 5 c), and after sequencing was correct, the plasmids were extracted and purified using a plasmid purification kit from Qiagen, to obtain high-quality plasmids for each recombinant expression vector.
EXAMPLE 3 preparation of TCR-T
Preparation of human activated T cells:
coating six-well plates with a coating solution containing 5. Mu.g/ml of an anti-CD 3 antibody and 5. Mu.g/ml of an anti-CD 28 antibody at room temperature for 2 to 4 hours, after blotting the coating solution, washing the plates 1 to 3 times with physiological saline, and adding 2% FBS-containing AIM-V medium for use; resuscitating human peripheral blood PBMC (HLA-02, 01, purchased from ALLCELLS) in a37 ℃ water bath, culturing the PBMC in an adherent manner for 2-4h, wherein nonadherent suspension cells are initial T cells, collecting the suspension cells into a15 ml centrifuge tube, centrifuging for 3min at 1200rmp, discarding supernatant, adding physiological saline, centrifuging for 3min at 1200rmp, discarding physiological saline, and repeating the steps; the washed primary T cells are then transferred to antibody-coated wells containing the medium to be used, incubated at 37 ℃ 5% in CO2 for 3-4 days before carrying out the subsequent experiments.
Electrically converting to prepare TCR-T cells expressing NY-ESO-1 TCR and NY-ESO-1 TCR + TGF-beta signal conversion receptors:
1) AIM-V medium was previously added to 19 wells of a 12-well plate at 2mL per well, followed by transferring into a cell incubator at 37 ℃ 5% for preheating in CO2 for 1 hour;
2) The electrotransfer formulation for a single dose per well was carried out according to table 7 below for a total of 19 wells:
TABLE 7
100μL Nucleocuvette TM Strip(μL)
Volume of Nucleofector TM solution 82
Electrotransformation make-up solution 18
3) Taking the obtained activated T cells into 19 EP tubes, adding 5X 106 cells into each EP tube, centrifuging for 5min at 1200rpm, discarding the supernatant, then resuspending the cells with 500. Mu.L of physiological saline, and repeating the centrifugation step to wash the cell pellet;
4) The electrotransfer liquid prepared in the step 2) is respectively added with plasmids pNB328-1682 ug, pNC328-1682 ug + pNB328-1648 ug, pNC328-1682 ug + pNB328-1650 3 ug, pNC328-1682 ug + pNB328-1652 ug, pNC328-1682 ug + pNB328-1654 ug, pNC328-1682 ug + pNB328-1656 ug, pNC328-1682 ug + pNB328-1658 ug, pNC328-1682 ug + pNB328-1660 ug, pNC328-1682 ug + pNB328-1662 ug pNC328-1682 μ g + pNB328-1664 μ g, pNC328-1682 μ g + pNB328-1666 μ g, pNC328-1682 μ g + pNB328-1668 3 μ g, pNC328-1682 3 μ g + pNB328-1670 μ g, pNC328-1682 3 μ g + pNB328-1672 μ g + pNB328-1674 μ g, pNC328-1682 3 μ g + pNB328-1676 μ g, pNC328-1682 3 μ g + pNB328-1678 μ g, pNC328-1682 3 μ g + NB328-1680 μ g, and NC empty vector 3 μ g + pNB empty vector 3 μ g, then standing at room temperature for less than 30 min;
5) Resuspending 19 tubes of the plasmid-containing electrotransfer prepared in 4) into activated T cells, 100. Mu.L per tube, carefully pipetting the cell resuspension into a LONZA 100. Mu.L electrotransfer cup, and placing the electrotransfer cup into a LONZA Nucleofector TM 2b, starting an electrotransfer program in the electrotransfer tank, wherein the electrotransfer program selects T-020;
6) After the electrotransfer was completed, the electric rotor was carefully removed, the cell suspension was aspirated and transferred to an EP tube, 200. Mu.L of preheated AIM-V medium was added to each tube, and then transferred to a 12-well plate of 1) containing preheated AIM-V medium, and cultured at 37 ℃ and 5% CO2, after 1 hour of culture, compound G150 (purchased from MedChemexpress) was added to a final concentration of 5. Mu.M, followed by further culture for 13 days during which passage was performed according to cell proliferation, and after 13 days, the cell number and cell survival rate of each electrotransfer sample were measured.
The cells expressing exogenous genes prepared by the method are named as TCR-T, SCR TCR-T, SCR2 TCR-T, SCR3 TCR-T, SCR TCR-T, SCR5TCR-T, SCR TCR-T, SCR7 TCR-T, SCR8 TCR-T, SCR TCR-T, SCR10 TCR-T, SCR TCR-T, SCR12 TCR-T, SCR TCR-T, SCR TCR-T, SCR TCR-T, RIID TCR 562 TCR-6262 zxft 624 TCR-T and Mock TCR-T, respectively. The obtained above-mentioned cytometric flow cytometer (Beckman Cytoflex) was used to detect the EGFP-positive cell fraction. For TCR-T, EGFP-positive cells can be considered as cells positive for expression of the NY-ESO-1 TCR gene. For cells co-transfected with TGF- β signaling receptor and TCR, EGFP-positive indicates that the PB transposase on which successful integration expression of TCR-EGFP depends is from the pNB vector expressing the gene encoding the TGF- β signaling receptor because the pNC vector does not contain a PB transposase expression cassette, and thus in cells co-transfected with TGF- β signaling receptor and TCR EGFP-positive cells can be considered to be dual integration expression of the TGF- β signaling receptor and the NY-ESO-1 TCR. The results are shown in FIG. 1 and Table 8:
TABLE 8
Group of cells EGFP Positive cell proportion (%)
Mock-TCR-T 0
TCR-T 44.22
SCR1 TCR-T 17.04
SCR2 TCR-T 15.10
SCR3 TCR-T 20.68
SCR4 TCR-T 31.69
SCR5 TCR-T 20.82
SCR6 TCR-T 18.83
SCR7 TCR-T 41.17
SCR8 TCR-T 27.13
SCR9 TCR-T 19.42
SCR10 TCR-T 21.39
SCR11 TCR-T 18.07
SCR12 TCR-T 16.65
SCR13 TCR-T 23.76
SCR14 TCR-T 29.58
SCR15 TCR-T 31.24
RIID2 TCR-T 23.31
RIID4 TCR-T 21.75
FIG. 1 shows fluorescence micrographs of TCR-T cells with the TCR-EGFP fusion gene transfected and SCR1-5 TCR-T cells with the TCR-EGFP fusion gene + TGF- β signal transduction receptor gene co-transfected. EGFP fluorescence brightness of TCR-T is brighter than that of SCR1-5 TCR-T, probably because TCR-T is prepared by single plasmid, and the single plasmid simultaneously comprises exogenous TCR-EGFP fusion gene and PB transposase expression frame; SCR1-5 TCR-T needs two plasmids to be prepared by cotransformation, wherein exogenous TCR-EGFP fusion genes and PB transposase expression frames need two vectors of pNC and pNB to be provided respectively.
Example 4 killing Effect of TCR-T expressing TGF- β Signaling receptor SCR1-5 on target cells
HLA-02 of NY-ESO-1 expression positive type A375 (all purchased from American type culture Collection ATCC) is selected as a target cell, and real-time unmarked cell function analyzer (RTCA) of the company Aisen is used for detecting the in vitro killing activity of the TCR-T, SCR TCR-T, SCR TCR-T, SCR3 TCR-T, SCR TCR-T, SCR TCR-5 TCR-T, RIID2 TCR-T and RIID4 TCR-T cells prepared in example 3, and the specific steps are as follows:
(1) Zero setting: adding 50 mul DMEM or 1640 culture solution into each well, putting into an instrument, selecting step 1, and zeroing;
(2) Target cell plating: the melanoma cells A375 are paved in a plate containing a detection electrode according to the proportion of 104 cells/50 mu l per hole, are placed for a plurality of minutes, are placed in an instrument after the cells are stabilized, and step 2 is started to culture the cells;
(3) Adding effector cells: after the target cells were cultured for 8-48h until the Cell Index (Cell Index) reached 1.0, step 2 was suspended, effector cells were added in 50. Mu.l/well, the number of effector cells was added at an effective target ratio of 1.5: 1 in the proportion of positive cells in Table 1, step 3 was started, and the Cell proliferation curve was observed after further co-culture for more than 120 h.
The results are shown in FIGS. 2 and 3. FIGS. 2 and 3 show the killing effect of SCR1-5 TCR-T, RIID2 TCR-T and TCR-T on target cells and the killing effect of SCR1-5 TCR-T, RIID TCR-T and TCR-T on target cells, respectively. The killing effect is observed for a long time by setting the effective target ratio to be 1.5: 1. Both FIGS. 2 and 3 show that SCR4 TCR-T and SCR5TCR-T have shown significantly better target cell killing than TCR-T, SCR1/2/3 TCR-T, RIID TCR-T and RIID4 TCR-T shortly after the addition of the effector cell time point (24 h). After the killing time reaches 120h, the killing effect of SCR1-5 TCR-T on the target cell A375 is still more obvious compared with TCR-T, wherein the killing effect of SCR1/2/3 TCR-T is close, the killing effect of SCR4 TCR-T is better than that of SCR1/2/3 TCR-T, and the killing effect of SCR5-TCR-T is obviously better than that of SCR1-4 TCR-T and TCR-T, and is optimal in a plurality of effector cells. FIG. 2 shows that the killing effect of RIID2 TCR-T is close to that of TCR-T, and is slightly lower than that of TCR-T; FIG. 3 shows that the killing effect of RIID4 TCR-T is slightly better than TCR-T, but lower than SCR1/2/3 TCR-T, and also significantly lower than SCR4 TCR-T and SCR5TCR-T. The above results show that the signal conversion receptor SCR1-5 of the present invention can still significantly increase the killing level of effector T cells to tumor target cells under the condition of low target ratio, and the enhancing effect still exists when the killing time is long.
Example 5 specific killing of TCR-T on target cells at low potency targets by expression of TGF- β Signaling receptor SCR5 Fruit (A. A. B. D. B
Referring to example 4, the same procedure as in example 4 was followed, using A375 as target cells, selecting TCR-T, SCR5TCR-T, RIID2 TCR-T and RIID4 TCR-T as effector cells, further reducing the effector-target ratio to 1: 1, and comparing the killing level of A375 cells by several effector cells. The 1: 1 low-efficiency target ratio long-time observation aims at simulating a scene of killing tumors by T cells in a real environment in vivo.
The results are shown in FIGS. 4 and 5. FIG. 4 shows that shortly after the time point of effector cell addition (just after 40 h), the killing effect of RIID2 TCR-T has been shown to be superior to TCR-T, while the killing effect of SCR5TCR-T is significantly superior to the former two. After the killing detection time length exceeds 120h, the killing effect of RIID2 TCR-T is still better than that of TCR-T, and the killing effect of SCR5TCR-T is still better than that of the former two. Similarly, FIG. 5 shows that the killing effect of RIID4 TCR-T has been shown to be superior to TCR-T shortly after the time point of effector cell addition (just after 40 h), while the killing effect of SCR5TCR-T is significantly superior to the first two. When the killing detection time length exceeds 120h, the killing effect of RIID4 TCR-T is still better than that of TCR-T, and the killing effect of SCR5TCR-T is also better than that of the former two. The above results show that even under the conditions of extremely low effective target ratio and long killing time, the signal conversion receptor SCR5 of the invention can still effectively improve the killing level of effector T cells to tumor target cells.
Example 6 killing Effect of TCR-T expressing TGF- β Signaling receptor SCR6-10 on target cells
The in vitro killing activity of the TCR-T, SCR6 TCR-T, SCR7 TCR-T, SCR TCR-T, SCR TCR-T and SCR10 TCR-T prepared in example 3 on A375 cells was detected, the effective-to-target ratio was set to be 1.5: 1 according to the EGFP positive cell ratio, and the specific detection procedure was the same as that in example 4. The results are shown in FIG. 6. Shortly after the time point (24 h) when the effector cells were added, SCR10 TCR-T had shown a significantly better target cell killing effect than TCR-T, SCR6/7/8/9 TCR-T. After the killing time reaches 90h, the killing effect of SCR6-10 TCR-T on the target cell A375 is still more obvious compared with TCR-T, wherein the killing effect of SCR6/7/8 TCR-T is close, the killing effect of SCR9 TCR-T and SCR10 TCR-T is better than that of SCR6/7/8 TCR-T, and the killing effect of SCR10 TCR-T is optimal in a plurality of effector cells. The above results show that the signal conversion receptor SCR6-10 of the present invention can still significantly increase the killing level of effector T cells to tumor target cells under the condition of low target ratio, and the enhancing effect still exists when the killing time is longer.
Example 7 killing of target cells by TCR-T expressing TGF- β Signaling receptor SCR10 at low target ratios Effect
Referring to example 4, the same procedure as in example 4 was followed using A375 as target cells, selecting TCR-T, SCR10 TCR-T and RIID2 TCR-T as effector cells, further reducing the effector-target ratio to 0.5: 1, and comparing the level of killing of A375 by several effector cells. The 0.5: 1 inefficient target ratio long-term observation aims at simulating a scene that T cells kill tumors under a real environment in vivo.
The results are shown in FIG. 7. FIG. 7 shows that after 80h, the killing effect of RIID2 TCR-T has been shown to be superior to TCR-T, while the killing effect of SCR10 TCR-T is significantly superior to the former two. After the killing detection time is 140 hours, the killing effect of the RIID2 TCR-T is still better than that of the TCR-T, and the killing effect of the SCR10 TCR-T is obviously better than that of the TCR-T. The results show that the signal conversion receptor SCR10 can still effectively improve the killing level of effector T cells on tumor target cells even under the conditions of the effective target ratio as low as 0.5: 1 and longer killing time.
Example 8 killing Effect of TCR-T expressing TGF- β Signaling receptor SCR11-15 on target cells
The in vitro killing activity of the TCR-T, SCR-15 TCR-T prepared in example 3 on A375 cells was determined, the efficacy-to-target ratio was set at 1.5: 1 according to the EGFP positive cell ratio, and the specific detection procedure was the same as that of example 4. The results are shown in FIG. 8. Shortly after the time point (24 h) when the effector cells were added, SCR11/12/14/15 TCR-T had shown a significantly better target cell killing effect than TCR-T. After the killing time reaches 90h, compared with TCR-T, SCR11-15 TCR-T has more obvious killing effect on target cells A375, wherein the killing effect of SCR11/12 TCR-T is close to and better than that of SCR13 TCR-T; the killing effect of SCR14/15 TCR-T is better than that of SCR11/12 TCR-T, and the killing effect of SCR15 TCR-T is the best among several effector cells. The above results show that the signal conversion receptor SCR11-15 of the present invention can still significantly increase the killing level of the effector T cells to the tumor target cells under the condition of low target ratio, and the enhancing effect still exists when the killing time is longer.
Example 9 killing of target cells by TCR-T expressing TGF- β Signaling receptor SCR15 at low target ratios Effect
Referring to example 4, A375 was selected as the target cells, TCR-T, SCR TCR-T and RIID4 TCR-T were selected as the effector cells, the effector-target ratio was further reduced to 0.75: 1, and the killing level of A375 cells by several effector cells was compared, which was performed as in example 4. The 0.75: 1 inefficient target ratio was observed over a long period of time in a scenario intended to mimic the killing of tumors by T cells in real in vivo environments.
The results are shown in FIG. 9. FIG. 9 shows that shortly after addition of effector cells, the killing effect of RIID4 TCR-T has been shown to be superior to TCR-T, while the killing effect of SCR15 TCR-T started before 60h to be significantly superior to the first two. After the killing detection time is 100 hours, the killing effect of the RIID4 TCR-T is still better than that of the TCR-T, and the killing effect of the SCR15 TCR-T is obviously better than that of the TCR-T. The results show that the signal conversion receptor SCR15 can still effectively improve the killing level of effector T cells on tumor target cells even under the conditions of the effective target ratio as low as 0.75: 1 and longer killing time.
Example 10: TGF-beta signal transduction receptor for increasing secretion level of TCR-T cytokine
TGF-. Beta.1 factor (purchased from R & D SYSTEMS, cat. No.: 240-B-002/CF) was added to the TCR-T and SCR1-5 TCRT cell culture SYSTEMS prepared in example 3 to a final concentration of 2.0ng/mL, 3 parallel sample wells were provided for each group of cells, and another group of TCR-T was not treated with TGF-. Beta.1 factor. After 24 hours of treatment, IFN-. Gamma.secretion levels were measured using HTRF IFN-. Gamma.detection kit (Cisbio Human IFN gamma kit cat # 62 HIFGPET) according to the protocol. The results are shown in FIG. 10.
FIG. 10 shows that IFN- γ secretion levels were significantly reduced in the TCR-T group treated with TGF-. Beta.1 as compared to the TCR-T group not treated with TGF-. Beta.1, indicating that TGF-. Beta.receptor naturally present on the surface of T cells produces an inhibitory effect on T cell activation upon binding of TGF-. Beta.1 factor. After SCR1-5 TCR-T is treated by TGF-beta 1 factor, the secretion level of IFN-gamma is obviously improved compared with the TCR-T group treated by TGF-beta 1 or compared with the TCR-T group not treated by TGF-beta 1. This indicates that immune effector cells, such as TCR-T cells, upon introduction of the signal transduction receptors of the invention, are effective in increasing the level of T cell activation upon binding of the corresponding ligand, such as TGF-. Beta.1 factor, and that this increase can counteract and negate the inhibitory effect that the corresponding ligand, such as TGF-. Beta.1, has on the cells following binding to its own natural inhibitory receptors on their surfaces.
Example 11: TGF-beta signal conversion receptor promotes in vivo tumor suppression levels of TCR-T
Detection of TCR-T, RIID2 TCR-T, RIID TCR-T and SCR5TCR-T in immunodeficient mouse model for tumor growth following tumor cell line A375 in vivo tumor formationInhibiting effect. Immunodeficient mouse NSG (purchased from The Jackson Laboratory) was selected for in vivo tumorigenesis of The A375 cell line. NSG mice are bred in a standard SPF animal room for two weeks to adapt to the environment before being inoculated with tumor cells, 12h circadian rhythm alternation control is provided, the environment temperature is 20-24 ℃, and the humidity is 45-65%. A375 cells were cultured in DMEM medium and 10% fetal calf serum to 80-90% confluence, counted and calculated for survival, and 100 μ L of cells containing 1X 10 was subcutaneously injected to the left side of each mouse after isoflurane anesthesia 7 PBS cell suspension of a375 live cells to minimize pain sensation in mice. After the mice had developed tumors, tumor-bearing mice were randomly grouped into 5 groups in total, n =6, into a control group (a 375), a TCR-T group, a RIID2 group, a RIID4 group, and an SCR5 group. The cells prepared in example 3 were used for reinfusion. After the cells were screened through a 70 μm sieve, they were resuspended in PBS. According to the EGFP-positive rate reported in Table 8, the TCR-T was 44.22%, which was approximately 2-fold higher than the EGFP-positive cell ratio in RIID2 TCR-T, RIID TCR-T and SCR5TCR-T, and the cell dose of the TCR-T group was 1X 10 7 The cell dose of 2X 10 cells resuspended in 200. Mu.L PBS, RIID2 TCR-T, RIID TCR-T and SCR5TCR-T groups 7 Cells, resuspended in 200 μ Ι _ PBS; control groups were given 200 μ L PBS each. The drug was administered by tail vein injection at three time points D0, D1 and D2, respectively. Mice were observed for a trend of tumor size over time.
The results are shown in FIG. 11. Tumor proliferation was slowed in TCR-T tumor-bearing mice compared to the control group without cell reinfusion, tumor proliferation was further inhibited in rid 2 and rid 4 groups compared to TCR-T, and tumor proliferation was slightly slower in rid 4 group than in rid 2 group. The inhibition of tumor proliferation in the SCR5 group was most pronounced in several groups of mice, with tumor proliferation rates further inhibited than in the rid 2 and rid 4 groups. The results show that the immune effector cells expressing the TGF-beta signal conversion receptor SCR5, such as TCR-T, can have obviously improved inhibition effect on tumor proliferation in vivo.
Although specific embodiments of the invention have been described in detail, those skilled in the art will appreciate that the invention has many modifications and variations. Many modifications and variations of those details may be made in light of the overall teachings of the disclosure, and such variations are within the scope of the invention. The full scope of the invention is given by the appended claims and any equivalents thereof.

Claims (14)

  1. An isolated fusion protein comprising:
    an extracellular domain of a receptor for an immunosuppressive cytokine or a functional fragment or mutant thereof that retains a specific binding to an immunosuppressive cytokine;
    a transmembrane region or a mutant thereof; and
    the intracellular domain of the costimulatory signaling molecule or a functional fragment or mutant thereof that retains the biological function of the costimulatory signaling molecule to deliver a costimulatory signal to activate an immune cell.
  2. The fusion protein of claim 1, wherein the immunosuppressive cytokine comprises a tumor cell or tumor stromal cell membrane protein and a tumor cell or tumor stromal cell secreted protein.
  3. The fusion protein of claim 2,
    the tumor cell or tumor stroma cell membrane protein comprises PD-L1/PD-L2, fasL, B7-H4, crry and HLA-G;
    the tumor cell or tumor stromal cell secreted protein comprises IL-13, IL-4, TGF-beta, IL-6, IL-8, IL-10, CCL21, IDO and VEGF.
  4. The fusion protein of claim 3, wherein the receptor for TGF- β comprises TGF- β receptor 1, TGF- β receptor 2, and TGF- β receptor 3, preferably TGF- β receptor 1 and TGF- β receptor 2.
  5. The fusion protein of any one of claims 1-4, wherein the costimulatory signal molecule comprises one or more of CD28, CD134 (OX 40), CD137 (4-1 BB), LCK, ICOS, DAP10, siglec-9, siglec-10, siglec-15, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, IL-R2, IL-4R, IL-7R, IL-10R, IL-12R, IL-15R, IL-21R, CD and CD40.
  6. The fusion protein of any one of claims 1-5, wherein the transmembrane region comprises any one or more of the transmembrane regions from CD28, CD8, CD134 (OX 40), CD137 (4-1 BB), LCK, ICOS, DAP10, siglec-9, siglec-10, siglec-15, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, IL-2Rbeta, IL-2Rgamma, IL-4Ralpha, IL-7Ralpha, IL-10R, IL-12R, IL-15R, IL-21R, CD and CD40.
  7. The fusion protein of claim 6, wherein the transmembrane region is from an IL-7Ralpha transmembrane region or mutant thereof;
    preferably, the amino acid sequence of the IL-7Ralpha transmembrane region is shown as SEQ ID NO. 27; more preferably, the coding sequence of the IL-7Ralpha transmembrane region is shown in SEQ ID NO 28;
    preferably, the amino acid sequence of the mutant of the IL-7Ralpha transmembrane region is shown as any one of SEQ ID NO. 29-60; more preferably, the coding sequence of said mutant of the IL-7Ralpha transmembrane domain is as shown in any one of SEQ ID NO 61-94.
  8. The fusion protein of claim 7, wherein the extracellular region of the fusion protein is an extracellular region of TGF- β receptor 1, and the intracellular domain of the co-stimulatory molecule of the fusion protein is any one selected from the group consisting of an IL-7Ralpha intracellular region, a CD28 intracellular region in tandem with a 4-1BB intracellular region, a CD28 intracellular region, a 4-1BB intracellular region, a CD28 intracellular region in tandem with an OX40 intracellular region, a 28-IL2RB-z (YXXQ) intracellular region, a 28-DeltaIL2RB-z (YXXQ) intracellular region, an IL2RB-z (YXXQ) intracellular region, and a DeltaIL2RB-z (YXXQ) intracellular region; preferably, the amino acid sequence of the fusion protein is shown in any one of SEQ ID NO 95-350.
  9. The fusion protein of claim 7, wherein the extracellular region of the fusion protein is an extracellular region of TGF- β receptor 2, and the intracellular domain of the costimulatory molecule of the fusion protein is any one selected from the group consisting of an IL-7Ralpha intracellular region, a CD28 intracellular region in tandem with a 4-1BB intracellular region, a CD28 intracellular region, a 4-1BB intracellular region, a CD28 intracellular region in tandem with an OX40 intracellular region, a 28-IL2RB-z (YXXQ) intracellular region, a 28-DeltaIL2RB-z (YXXQ) intracellular region, an IL2RB-z (YXXQ) intracellular region, and a DeltaIL2RB-z (YXXQ) intracellular region; preferably, the amino acid sequence of the fusion protein is any one of SEQ ID NO 351-606, SEQ ID NO 1647, SEQ ID NO 1649, SEQ ID NO 1651, SEQ ID NO 1653, SEQ ID NO 1655, SEQ ID NO 1657, SEQ ID NO 1659, SEQ ID NO 1661, SEQ ID NO 1663, SEQ ID NO 1665, SEQ ID NO 1667, SEQ ID NO 1669, SEQ ID NO 1671, SEQ ID NO 1673 and SEQ ID NO 1675.
  10. The fusion protein of claim 7, wherein the extracellular region of the fusion protein is an extracellular region of TGF- β receptor 3, and the intracellular domain of the costimulatory molecule of the fusion protein is any one selected from the group consisting of an IL-7Ralpha intracellular region, a CD28 intracellular region in tandem with a 4-1BB intracellular region, a CD28 intracellular region, a 4-1BB intracellular region, a CD28 intracellular region in tandem with an OX40 intracellular region, a 28-IL2RB-z (YXXQ) intracellular region, a 28-DeltaIL2RB-z (YXXQ) intracellular region, an IL2RB-z (YXXQ) intracellular region, and a DeltaIL2RB-z (YXXQ) intracellular region; preferably, the amino acid sequence of the fusion protein is shown in any one of SEQ ID NO 607-862.
  11. A polynucleotide molecule selected from the group consisting of:
    (1) A polynucleotide molecule encoding the fusion protein of any one of claims 1-10; and
    (2) (1) the complement of the polynucleotide molecule;
    preferably, the polynucleotide molecule comprises a sequence selected from any one of SEQ ID NO 863-1630, SEQ ID NO 1648, SEQ ID NO 1650, SEQ ID NO 1652, SEQ ID NO 1654, SEQ ID NO 1656, SEQ ID NO 1658, SEQ ID NO 1660, SEQ ID NO 1662, SEQ ID NO 1664, SEQ ID NO 1666, SEQ ID NO 1668, SEQ ID NO 1670, SEQ ID NO 1672, SEQ ID NO 1674 and SEQ ID NO 1676, or is a complement of any one of the polynucleotide molecules shown.
  12. A nucleic acid construct comprising the polynucleotide molecule of claim 11; preferably, the nucleic acid construct is a vector, more preferably an expression vector.
  13. A genetically engineered cell expressing the fusion protein of any one of claims 1-10, and/or carrying a coding sequence for the fusion protein;
    preferably, the cell is an immune cell; more preferably, T cells;
    preferably, the cell further expresses a CAR, or carries a coding sequence for a CAR;
    preferably, the cell also expresses an exogenous TCR, or carries the coding sequence of an exogenous TCR.
  14. Use of the fusion protein of any one of claims 1-10, the polynucleotide molecule of claim 11, the nucleic acid construct of claim 12, and the genetically engineered cell of claim 13 in the preparation of a medicament for treating or preventing cancer.
CN202180039676.XA 2020-06-01 2021-05-31 Signal transduction receptors and uses thereof Pending CN115698038A (en)

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