106 results about "NKG2D" patented technology

The invention discloses an E-CAR-NK cell for treating systemic lupus erythematosus and application of the E-CAR-NK cell, and belongs to the technical field of genetic engineering.A CD19 and BCMA double-target E-CAR is adopted for modifying an NK cell, and the E-CAR-NK cell is obtained; the CD19 and BCMA double-target E-CAR is composed of the following modules: a CD8 signal peptide, a CD19 scFv, a Linker, a BCMA scFv, a CD8 Hinge region, an NKG2D transmembrane region, a CD3 epsilon signal transduction region, a DAP12 costimulatory region, a 2B4 costimulatory region, and a CD3 zeta signal transduction region. Compared with a common CAR-NK cell, the E-CAR-NK cell disclosed by the invention has higher antigen sensitivity and better continuous killing ability, so that the treatment effect of SLE is improved, and the E-CAR-NK cell has lower side effects and higher safety.

According to the engineering immune cell for immunotherapy and the preparation method thereof, the engineering immune cell is more durable in a host organism, and the transplantation survival rate is high.

The invention discloses a culture medium composition for culturing GDT cells and a culture method, and belongs to the field of biological medicine. Aiming at the problems of low in-vitro amplification efficiency and poor function of the existing GDT cells, the invention provides the culture medium composition for culturing the GDT cells, the culture medium composition comprises an induction culture medium and a proliferation culture medium, the induction culture medium comprises a basic culture medium and an activating agent, and the activating agent comprises IL-7 and paclitaxel; the proliferation culture medium comprises a basic culture medium, IL-18 and a metabolism regulator. An STAT5 pathway is activated through IL-7, and amplification of the Vdelta2 subgroup is specifically promoted; on the other hand, through the cooperation of the IL-7 and the IL-18, the IL-18 activates the expression of the NKG2D, and the IL-18 and the IL-7 jointly enhance the cytotoxicity; efficient amplification and function enhancement of the GDT cells are achieved; the killing rate of PD-L1 + tumor cells is larger than 80%, the motility rate after cryopreservation recovery is larger than 90%, the function maintenance rate is larger than 85%, and the cell line is suitable for tumor immunotherapy, infectious disease treatment and cell drug development.

Multi-specific binding proteins that bind to and kill human cancer cells are described, as well as pharmaceutical compositions and therapeutic methods useful for the treatment of cancer. The cancer can be B-cell maturation antigen (BCMA)-expressing cancer. The multi-specific binding proteins provided herein exhibit high potency and maximum lysis of target cells compared to anti-BCMA monoclonal antibodies.

Several embodiments of the methods and compositions disclosed herein relate to immune cells that are gene edited (e.g., using Crispr / Cas) to modulate, reduce, or otherwise eliminate expression of one or more endogenous genes. In several embodiments, the edited cells are engineered to express chimeric antigen receptors that target tumor antigens (e.g., CD19, ligands of the NKG2D receptor, CD70, and / or BCMA, etc.). In several embodiments, the editing enhances one or more aspects of immune cell efficacy in cellular immunotherapy, including cytotoxicity (e.g., ADCC) and / or persistence.

The invention discloses a sequential anti-tumor strategy based on combination of improved tumor microenvironment (TME) and CAR T and application of the sequential anti-tumor strategy, belongs to the technical field of biological medicine, and particularly relates to a sequential anti-tumor strategy based on combination of improved tumor microenvironment (TME) and CAR T. According to the strategy, tumor fibrosis and an acidic microenvironment are improved at the same time by applying a tumor-associated fibroblast (CAF) targeted drug and a proton pump inhibitor, and then sequential injection of CAR T is performed for combined treatment of solid tumors. Wherein the CAF targeting drug is selected from all-trans retinoic acid, calcipotriol, losartan and minetil, the proton pump inhibitor is selected from lansoprazole, omeprazole, pantoprazole, rabeprazole and esomeprazole, and the CAR T receptor or ligand is selected from mesothelin, protein tyrosine kinase 7, NKG2D, CD47, B7-H3, MUC1 and HER2; the sequential administration strategy firstly destroys a compact physical barrier of a tumor microenvironment, improves acidity of a tumor site, ensures infiltration, survival and proliferation of CAR T at the tumor site, and then utilizes specificity of CAR T to inhibit growth of solid tumors in a targeted manner, especially triple negative breast cancer with a high malignancy degree.

Multi-specific binding proteins that bind to and kill human cancer cells expressing epidermal growth factor receptor 2 (HER2 or ErbB2), but does not kill non-cancerous healthy human cells expressing HER2 are described, as well as pharmaceutical compositions and therapeutic methods useful for the treatment of HER2 expressing cancer. The invention also relates to multi-specific binding proteins that trigger CD8+ T cell killing of tumor cells.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-NKG2D nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing NKG2D protein, immunizing alpaca to obtain immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the PBMC, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), obtaining an antibody gene segment through PCR (polymerase chain reaction), and constructing the antibody gene segment on a phage expression vector; the method comprises the following steps: carrying out electrotransformation on a vector with an antibody gene segment to an SS320 electrotransformation competent state to obtain an NKG2D immune antibody library, carrying out two rounds of NKG2D protein panning and Clone ELISA detection to obtain an antibody combined with NKG2D protein, carrying out sequencing analysis, selecting a unique sequence, and carrying out high-throughput expression through a mammalian cell high-throughput expression system to obtain an antibody protein. And carrying out FACS detection by using an NKG2D high-expression cell line to finally obtain the antibody combined with the NKG2D. The method disclosed by the invention is simple and quick to operate; a mammalian cell expression system induces high-efficiency expression of the antibody, processing modification can be performed after translation, and the activity of the antibody is closer to that of a natural antibody.

The present invention relates to a method for preparing immune response cells, immune response cells thereof, pharmaceutical compositions, kits, uses thereof and methods of treatment thereof. Specifically, the invention provides a method for preparing an immune response cell, and the method comprises the following steps: (a) carrying out gene modification on an immune cell to express an NKG2D polypeptide; wherein the immune cells have been activated by anti-CD3 and anti-CD28 antibodies or fragments thereof conjugated to a nanomatrix; or (b) activating an immune cell wherein the immune cell is genetically modified to express an NKG2D polypeptide, and wherein activation is performed by anti-CD3 and anti-CD28 antibodies or fragments thereof conjugated to a nanomatrix.

The application discloses a kind of nanobody specifically targeting human and murine NKG2D protein and its preparation method and application.The nanobody or antigen-binding fragment thereof of the application has three complementarity determining regions CDR1, CDR2 and CDR3;Wherein the amino acid sequences of CDR1, CDR2, CDR3 are as shown in SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4 respectively.The nanobody can simultaneously bind hNKG2D and mNKG2D with high affinity, and the bispecific nanobody E5 / 2-B12 based on the nanobody can specifically bind CEACAM5 positive tumor cells and NKG2D overexpression cells, and effectively activate NK cells, which can significantly inhibit tumor growth and prolong survival in various tumor models, and has good application prospect in tumor immunotherapy.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-NKG2D nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing NKG2D protein, immunizing alpaca to obtain immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the PBMC, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), obtaining an antibody gene segment through PCR (polymerase chain reaction), and constructing the antibody gene segment on a phage expression vector; the method comprises the following steps: carrying out electrotransformation on a vector with an antibody gene segment to an SS320 electrotransformation competent state to obtain an NKG2D immune antibody library, carrying out two rounds of NKG2D protein panning and Clone ELISA detection to obtain an antibody combined with NKG2D protein, carrying out sequencing analysis, selecting a unique sequence, and carrying out high-throughput expression through a mammalian cell high-throughput expression system to obtain an antibody protein. And carrying out FACS detection by using an NKG2D high-expression cell line to finally obtain the antibody combined with the NKG2D. The method disclosed by the invention is simple and quick to operate; a mammalian cell expression system induces high-efficiency expression of the antibody, processing modification can be performed after translation, and the activity of the antibody is closer to that of a natural antibody.

The invention belongs to the technical field of cell culture, and particularly relates to an umbilical cord blood NK cell culture dish and a use method thereof. The culture dish is reasonable in design, the bone marrow microenvironment is simulated through the three-dimensional micro-scaffold, and the adhesion and proliferation efficiency of cord blood NK precursor cells is remarkably improved by combining biocompatible nanofibers (PLGA) and extracellular matrix proteins (fibronectin and heparin sulfate), so that gt; the amplification multiple is 1 * 10 < 4 >. The dynamic microfluidic gradient channel continuously provides cell factors (such as IL-15 and IL-12) and nutrition, maintains stable glucose concentration (gt, 3mM) and low lactic acid level (lt, 15mM), and prevents metabolic waste from inhibiting amplification. According to the invention, an anti-CD16 antibody and an NKG2D ligand (ULBP2) are fixed on the surface, ADCC effect and tumor targeted killing ability of NK cells are enhanced through a costimulatory signal (CD137L), and cytotoxicity (K562 killing rate) gt is enhanced; 80%.

The invention belongs to the field of biological medicine, and relates to a CAR-T cell targeting NKG2DL and secreting IL-15 and a preparation method thereof, the method comprises the step of infecting a T cell with a recombinant lentivirus, and the recombinant lentivirus contains a nucleic acid molecule of a chimeric antigen receptor encoding NKG2D and IL-15 or is prepared through an expression vector containing the nucleic acid molecule. The CAR-T cell expresses a chimeric antigen receptor targeting NKG2DL and can secrete a cell factor IL-15, the proliferation and effector functions of the CAR-T cell are promoted, and the killing activity on tumors is enhanced.

The invention includes a chimeric antigen receptor (CAR) specific for a natural-killer group 2, member D ligand (NKG2DL), compositions comprising the CAR, polynucleotides encoding the CAR, vectors comprising a polynucleotide encoding the CAR, and recombinant T cells comprising the CAR. The invention also includes methods of making a genetically modified T cell expressing a NKG2D-CAR wherein the expressed CAR comprises a NKG2D extracellular domain. The present invention also relates generally to the use of T cells engineered to express a CAR to treat cancer associated with NKG2D.

This disclosure provides compositions and methods of making cells, such as allogenic cells, engineered to evade host immune responses, particularly from T cells and natural killer cells, and populations thereof. The compositions and methods for making engineered cells include making genetic modifications that reduce or eliminate the expression of an NKG2D ligand or integrate sequences encoding HLA molecules, such as HLA-E or HLA-G. Such modifications can enhance the survival of the engineered cells within a host without impeding their ability to differentiate into functional cell types, such as cardiomyocytes.

The invention discloses application of a peripheral T lymphocyte index in preparation of a coronary heart disease risk assessment kit, the kit and a method. Prediction indexes of the kit comprise four T lymphocyte subgroups: CD3 + CD8 + CD28-, CD3 + CD8 + HLADR +, Tc1, CD3 + gamma delta + V delta 2 + NKG2D +. The invention finds that the four novel lymphocytes in the peripheral blood are risk factors of the coronary heart disease for the first time, and the four risk factors are combined as a combined prediction index. The kit has an optimal effect on evaluating the risk of the coronary heart disease, and has a wide application prospect in the aspect of predicting the risk of the coronary heart disease.

Provided are an engineered TIL cell having an improved tumor recognition and killing capability and a use thereof, relating to the technical field of genetic engineering and cell therapy. The surface of the engineered TIL cell comprises an exogenous membrane-bound DAP10-CD3ζ fusion protein, and the endoplasmic reticulum or Golgi apparatus of the engineered TIL cell comprises a retained fusion protein. Specifically, provided is the membrane-bound DAP10-CD3ζ fusion protein, so that the TIL cell can be activated by an activated fusion protein when recognizing a tumor cell by means of the NKG2D-NKG2DL pathway, so as to effectively kill the tumor cell. Further provided is the retained fusion protein, so that endogenous NKG2DL that is expressed when the TIL cell is activated can bind to the retained fusion protein so as to be retained in the cell, thereby avoiding mutual recognition and killing between the TIL cells and improving the viability of the TIL cells.

The invention provides application of CAR-NK (chimeric antigen receptor-natural killer) cells in preparation of drugs for treating autoimmune diseases, and the CAR-NK cells express a chimeric antigen receptor and a fusion protein of a targeted NKG2D ligand; the fusion protein comprises an IL-15R alpha and an IL-15, and the IL-15 and the IL-15R alpha are connected with each other. According to the CAR-NK cell disclosed by the invention, the specific recognition of MICA / B molecules on the surface of an over-activated T / B cell of an SLE patient is realized by highly expressing an NKG2D activated receptor, and the durability of an intracellular signal is enhanced so as to resist cell depletion; meanwhile, the super IL-15 fusion protein is expressed to prolong the in-vivo survival and function duration time, and the effect of treating the SLE is improved through the cooperation of the dual mechanisms.

Provided is the use of NKG2D-CD3 and NKG2D-CD16 cell adapter molecules in the removal of aging cells and the treatment of aging-cell-accumulation-related diseases, wherein the cell adapter molecules comprise an NKG2D extracellular domain, a connection segment, and a CD3 or CD16 binding domain. The death of aging cells having high expression of NKG2D ligand is efficiently induced by means of bridging aging cells and immune cells, and thus the cell adapter molecules of the present application can be used for removing aging cells expressing the NKG2D ligand and treating aging-related diseases.

The present invention relates to an enhanced chimeric antigen receptor and use thereof, including a nucleic acid molecule for encoding a chimeric antigen receptor containing an NKG2D transmembrane region, a corresponding chimeric antigen receptor, a carrier, an immune effector cell, a preparation method and a product thereof, a pharmaceutical composition, pharmaceutical use, and a tumor or cancer treatment method. The present invention has the advantages of high CAR expression efficiency, rapid NK cell proliferation, strong killing ability against tumor cells, high specificity, long duration of killing effect, and the like.

Disclosed herein are immune effector cells that are expanded and enriched for NKG2D expression and genetically modified to express chimeric antigen receptor (CAR) polypeptides that can be used with adoptive cell transfer to target and kill CD33-expressing and / or CD123-expressing cancers. In some embodiments, the immune effector cells are gamma-delta (γδ) T cells, Natural Killer (NK) cells, or a combination thereof.

The present invention relates to NK cells expressing dual-targeting chimeric antigen receptors for CD19 and CD22, and uses thereof. In the present invention, it was found that the delivery efficiency of a CAR expression vector and the production efficiency of CAR-NK cells are excellent when, among the cell surface proteins of PBMCs for producing CAR-NK cells (UCI-101), CD16 is expressed at 70% or less, natural killer group 2D (NKG2D) is expressed at less than 10%, CD57 is expressed at 30% or less, low-density lipoprotein receptor (LDLR) is expressed at 0.1% or more, and natural cytotoxicity triggering receptor 3 (NKp30) is expressed at less than 10%. In addition, optimal conditions for inducing NK cell differentiation and optimal conditions for transduction, under which the delivery efficiency of a CAR expression vector and the production efficiency of CAR-NK cells are excellent, were established, and CAR-NK cells produced by the method according to the present invention were found to exhibit an excellent antitumor effect in an animal model, and thus can be effectively used as a composition for preventing or treating diseases related to CD22 (or CD19) expression or diseases related to B cells.

Several embodiments of the methods and compositions disclosed herein relate to immune cells engineered to express cytotoxic chimeric receptors and various dosing regimens for administering such cells. In several embodiments, the immune cell expresses a chimeric receptor that targets a ligand of NKG2D on a tumor cell. In several embodiments, the cancer is a leukemia, e.g., acute myelogenous leukemia (e.g., recurrent / refractory acute myelogenous leukemia) or myelodysplastic syndrome. In several embodiments, the tumor is a solid tumor, such as breast cancer, cervical cancer, colorectal cancer, gastric cancer, head and neck cancer, hepatocellular cancer, lung cancer, melanoma, intrahepatic cholangiocarcinoma, or other liver tumor, such as a secondary metastatic tumor from colorectal cancer. In several embodiments, the immune cells are administered in combination with a therapeutic agent, such as an additional anti-cancer agent.

The application belongs to the technical field of biological medicine, and particularly relates to application of a KAT5 inhibitor in a medicine for enhancing the immunotherapy effect of natural killer cells, and a medicine for enhancing the immunotherapy effect of natural killer cells. The KAT5 inhibitor comprises at least one of Nu9056, TH1834 and MG149. The KAT5 inhibitor can inhibit the recruitment of a transcription factor SP1 to an ADAM10 promoter, reduce the transcriptional activity of ADAM10, reduce the expression of NKG2D ligands on the cell surface, and thus enhance the sensitivity of tumor cells to NK cell killing. The application first discloses a new mechanism in which KAT5 regulates an ADAM10-NKG2D signal axis through an epigenetic mechanism, and proves that the KAT5 inhibitor can significantly enhance the immunotherapy effect of NK cells in vitro and in vivo. The application provides a brand-new target and strategy for developing a new immunosensitizer and a combined cell therapy.

The invention discloses an anti-NKG2D nano antibody, a fusion type nano antibody, a nucleic acid molecule, an expression vector, a recombinant cell and application thereof, and relates to the technical field of biological medicine. The anti-NKG2D nano antibody provided by the invention can specifically recognize and bind to NKG2D, can effectively competitively bind to a natural ligand (NKG2DL) of the NKG2D, and has relatively high affinity with the NKG2D. In-vitro experiments show that the provided antibody can effectively bind and activate NK cells so as to generate a remarkable killing effect on tumor cells. Therefore, the nano antibody provided by the invention can be applied to preparation of drugs for preventing or treating human tumors. In addition, the invention also provides a corresponding expression vector, a recombinant cell and an NKG2D detection product, an NKG2D enriched product and an NKG2D purified product related to the anti-NKG2D nano antibody, and the NKG2D detection product, the NKG2D enriched product and the NKG2D purified product have a wide application prospect.

The present disclosure relates to the treatment of cancer. In particular, the present disclosure provides novel therapeutic antibodies that target MHC Class I Chain-Related Protein A (MICA) in a manner that reduces the ability of a tumor cell to escape immune clearance mediated by the natural killer group 2D (NKG2D) receptor.

The invention relates to the technical field of biological medicine, and particularly discloses a chimeric antigen receptor of a targeted NKG2D ligand, a nucleic acid molecule, an expression vector, a macrophage and a preparation method and application of the chimeric antigen receptor, the nucleic acid molecule, the expression vector and the macrophage, and an extracellular antigen binding domain is an NKG2D extracellular structural domain; the hinge region is a CD28 hinge region or a CD8a hinge region; the transmembrane / intracellular signal transduction structural domain is an FCGR1 transmembrane / intracellular signal transduction structural domain or a CD3z transmembrane / intracellular signal transduction structural domain. The chimeric antigen receptor provided by the invention specifically recognizes and is combined with NKG2DLs highly expressed in NKG2DL positive malignant tumors, and macrophages expressing the chimeric antigen receptor can directly swallow tumor cells, promote M1 type polarization, reprogram tumor microenvironment and induce recruitment and activation of T cells, so that effective treatment of the malignant tumors is realized.

Several embodiments disclosed herein relate to the compositions comprising engineered Natural Killer (NK) cells that express a chimeric receptor, the chimeric receptor imparting to the NK cells an enhanced ability to target specific cells, such as cancerous cells or those affected by an infectious disease. Several embodiments relate to NK cells that target cells expressing natural ligands of NKG2D, where the NK cells comprise transmembrane and / or signaling domains that lead to cytotoxic and / or cytolytic effects when the NK cells bind a target cell. Uses of NK cell compositions to treat diseases are also provided for in several embodiments.

A nucleic acid sequence is provided that encodes a chimeric protein comprising a ligand that comprises a naturally occurring or modified follicle stimulating hormone sequence, e.g., an FSHβ sequence, or fragment thereof, which ligand binds to human follicle stimulating hormone (FSH) receptor, linked to either (a) a nucleic acid sequence that encodes an extracellular hinge domain, a transmembrane domain, a co-stimulatory signaling region, and a signaling endodomain; or (b) a nucleic acid sequence that encodes a ligand that binds to NKG2D. The vector containing the nucleic acid sequence, the chimeric proteins so encoded, and modified T cells expressing the chimeric protein, as well as method of using these compositions for the treatment of FSHR-expressing cancers or tumor cells are also provided.

To provide pharmaceutical formulations and administration regimens for treating cancer comprising a multi-specific binding protein that binds to HER2, NKG2D and CD16.SOLUTION: The disclosure relates to pharmaceutical formulations for multispecific binding proteins having an scFv that binds to epidermal growth factor receptor 2 (ErbB2 or HER2), a Fab that binds to NKG2D, and an antibody Fc domain. Dosage regimens for such multispecific binding proteins and pharmaceutical formulations for the treatment of cancer, such as locally advanced or metastatic solid tumors, are also provided. The disclosure provides pharmaceutical formulations comprising multispecific binding proteins comprising a HER2-binding scFv, a Fab that binds to NKG2D and an antibody Fc domain, of which ingredients are optimized for the stability of the multispecific protein.SELECTED DRAWING: None