145 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.

The present disclosure relates to a ULBP6 binding protein that inhibits the interaction between ULBP6 and NKG2D, and a method of treating cancer with the ULBP6 binding protein.

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 relates to the technical field of tumor immunotherapy, in particular to a dual-targeting chimeric antigen receptor, a recombinant expression vector, an anti-tumor natural killer cell and application, and provides a PD1-DAP10-41BB-P2A-NKG2D-Ig4-CD28-41BB-CD3 zeta fusion receptor.The dual-targeting chimeric antigen receptor can target PD-L1 on the surface of a tumor cell, can also recognize NKG2DLs and can be used for preparing anti-tumor natural killer cells. The broad spectrum of the NK cells is greatly improved, and meanwhile, the immune escape of tumor cells is also reduced.

The invention discloses a quality control method and platform for NK cell preparation, and relates to the technical field of cell preparation.The method comprises the steps that first preset culture environment information, EZH2 inhibitor weight parts and IL-15 fusion protein weight parts are processed through an NKG2D expression quantity prediction model, NKG2D predicted expression quantity is generated, when the NKG2D predicted expression quantity is inconsistent with target NKG2D expression quantity, directional optimization is conducted, and the NKG2D predicted expression quantity is obtained; obtaining culture environment baseline information meeting the target NKG2D expression quantity; when the culture environment monitoring time sequence information is inconsistent with the culture environment baseline information, an NK cell preparation quality control early warning signal is generated. The technical problem that in the large-scale preparation process, due to the uncertainty of the culture environment, the immunocompetence of the NK cells possibly has large difference is solved, and the purposes of ensuring the stability and consistency of the environment in the preparation process through a real-time monitoring and automatic early warning mechanism and improving the immunocompetence of the NK cells are achieved. Therefore, the stability and the consistency of the immunocompetence of the NK cells are improved.

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.

Multi-specific binding proteins that bind to and kill human cancer cells expressing CD33 (Siglec-3) are described, as well as pharmaceutical compositions and therapeutic methods useful for the treatment of CD33 expressing cancer. The invention relates to multi-specific binding proteins that bind to human cancer cells expressing CD33 and exhibit high potency and maximum lysis of target cells compared to anti-CD33 monoclonal antibodies. The multi-specific binding proteins comprise a CD33-binding domain, an NKG2D-binding domain and a CD16-binding domain.

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 present invention provides a non-human animal expressing a human or chimeric (e.g. humanized) NKG2D protein and a method of using same.

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.

To provide truncated NKG2D chimeric receptors and uses thereof in natural killer cell immunotherapy.SOLUTION: A polynucleotide encoding a chimeric receptor comprises: (a) an extracellular receptor domain, comprising a peptide that binds native ligands of Natural Killer Group 2 member D (NKG2D), wherein the peptide that binds native ligands of NKG2D is a fragment of NKG2D, and wherein the fragment of NKG2D is encoded by a polynucleotide comprising SEQ ID NO. 2; and (b) an effector domain, comprising a transmembrane region and an intracellular signaling domain, wherein the intracellular signaling domain comprises CD3ζ, and wherein the CD3ζ is encoded by a polynucleotide comprising SEQ ID NO. 13.SELECTED DRAWING: None

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 present invention belongs to the technical field of antibody preparation, and specifically relates to an anti-NKG2D nano antibody and its preparation method and application. The method of the present invention is as follows: construct and express NKG2D protein, immunize alpaca, obtain immune alpaca PBMC cells, extract RNA of PBMC cells, reverse transcribe into cDNA, obtain antibody gene fragments by PCR, and construct them on phage expression vectors; the obtained vector with antibody gene fragments is electrotransformed into SS320 electrotransformation competent state to obtain NKG2D immune antibody library, through 2 rounds of NKG2D protein panning, through Clone ELISA detection, obtain antibodies bound to NKG2D protein, after sequencing analysis, select unique sequences, and perform high-throughput expression through mammalian cell high-throughput expression system to obtain antibody protein, perform FACS detection with NKG2D high-expression cell line, and finally obtain antibodies bound to NKG2D. The method of the present invention is simple and fast to operate; and the mammalian cell expression system induces efficient expression of antibodies, which can be processed and modified after translation, and its activity is closer to natural antibodies.

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.