71 results about "NKG2D" patented technology

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.

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 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 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.

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 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.

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.

Described herein are pharmaceutical formulations comprising multi-specific binding proteins that bind to NKG2D, CD16, and epidermal growth factor receptor (EGFR); and therapeutic uses of the multi-specific binding proteins and pharmaceutical formulations thereof for treating a disease, for example, cancer, in a patient in need thereof.

The invention discloses a biomarker for pregnancy infection detection and an application method of the biomarker, and belongs to the technical field of pregnancy infection detection.According to the biomarker and the application method, multi-dimensional evaluation of the maternal infection state is achieved by combining immune activation, neurodevelopment risk and placenta function markers. According to the comprehensive detection mode, potential threats can be recognized earlier, limitation of a single index is avoided, and more accurate early warning signals are provided for clinic. Meanwhile, based on a clear high-risk threshold definition, a doctor can quickly lock a high-risk pregnant woman group and start targeted intervention in time, so that early-stage protection on healthy development of a fetus is enhanced, comprehensiveness and reliability of infection risk detection during pregnancy are remarkably improved, immune abnormality of a maternal-fetal interface is directly adjusted by blocking an NKG2D signal channel in a targeted manner, and the risk of infection caused by infection during pregnancy is reduced. The occurrence risk of the neurodevelopment disorder of the offspring is remarkably reduced, the maternal and infant health guarantee level is practically improved, and the effectiveness and operability of intervention measures are further enhanced.

The invention discloses a chimeric antigen receptor NK cell overexpressing a GPR133 gene, and a preparation method and application thereof, and belongs to the technical field of biological medicine. The preparation method comprises the following steps: (1) respectively mixing a pLL3.7-GPR133 plasmid with a packaging plasmid psPAX2, mixing a pLL3.7-NKG2D-CAR plasmid with a packaging plasmid psPAX2, mixing a pLL3.7-NKG2D-CAR plasmid with a packaging plasmid pMD2. G with a proper amount of an Opti-MEM culture medium; (2) mixing PEI with a proper amount of Opti-MEM culture medium, and incubating at room temperature; (3) adding the PEI-Opti-MEM solution in the step (2) into the Opti-MEM culture medium containing the plasmids in the step (1), and standing at room temperature; (4) adding the mixture obtained in the step (3) into 293T cells, uniformly mixing, and incubating at 37 DEG C; (5) collecting viruses from the cell supernatant, so as to respectively obtain a pLL3.7-NKG2D-CAR virus and a pLL3.7-GPR133 virus; and (6) co-transfecting an NK92 cell with the pLL3.7-NKG2D-CAR virus and the pLL3.7-GPR133 virus, so as to obtain the chimeric antigen receptor NK cell of the over-expressed GPR133 gene. According to the present invention, the GPR133 gene is subjected to overexpression in the CAR-NK cell for the first time, such that the biological function of the CAR-NK cell is significantly enhanced so as to improve the tumor cell killing ability of the CAR-NK cell.

The application provides application of psoralen in activating NK cells. The psoralen provided by the application is a coumarin component extracted from traditional Chinese medicine psoralea; through in-vitro cell experiment verification, the psoralen can activate NK-92 to secrete IFN-gamma, Granzyme B and Perforion, and activate surface protein NKG2D of NK-92; meanwhile, the psoralen also has the same effect on a cell group containing NK cells; through in-vivo experiment verification, the psoralen helps to improve the total amount of NK cells in the mouse spleen, inhibit tumor growth, and improve the expression amount of IFN-gamma, Granzyme B, Perforion, NKG2D, NKP46 and CD107a in a tumor infiltration environment. In summary, the small-molecule compound psoralen has good effect in activating NK cells.

This invention belongs to the field of biotechnology, specifically relating to a high-affinity anti-tumor NK cell, its preparation method, and its application. This invention designs NK92 cells transfected with a haPD1 high-affinity chimeric conversion receptor, wherein the high-affinity chimeric conversion receptor includes the extracellular segment of haPD-1, the transmembrane segment of CD28, the intracellular segment of DAP10, and the intracellular segment of CD3ζ. This invention prepares haChR3-NK92 cells through the construction of a recombinant lentiviral vector, lentiviral packaging, and lentiviral transfection of NK92 cells. In this invention, haPD-1 serves as the extracellular recognition domain of the CAR structure, specifically binding to PD-L1 on tumor cells to achieve a stronger tumor-killing effect. The co-stimulatory molecule CD28 serves as the transmembrane region to transmit extracellular information into the cell, integrating the adaptor protein DAP10 of the NK cell surface activator receptor NKG2D into the cell, and further embedding the intracellular segment CD3ζ commonly used in CAR design to jointly promote NK cell activation. The preparation method of this invention can successfully construct haChR3-NK92 cells, achieve the expression of the target plasmid, and be applied in anti-tumor drug research.

The application discloses a sequential anti-tumor strategy based on improvement of tumor microenvironment (TME) combined with CAR T and application thereof, and belongs to the technical field of biological medicine, which simultaneously improves tumor fibrosis and acid microenvironment by applying tumor-related fibroblast (CAF) targeting drugs and proton pump inhibitors, and sequentially injects CAR T combined therapy for solid tumors, wherein the CAF targeting drugs are selected from tretinoin, calcipotriol, losartan and minnelide, the proton pump inhibitors are 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 first destroys the dense physical barrier of the tumor microenvironment, improves the acidity of the tumor site, ensures the infiltration, survival and proliferation of CAR T at the tumor site, and then targets and inhibits the growth of solid tumors, especially high-malignancy triple-negative breast cancer, by using the specificity of CAR T.

The present invention relates to the field of cell drugs, specifically to a CAR-T cell drug that simultaneously targets two tumor antigens and its application. The tumor is solid glioma, and the CAR-T cell drug can reduce the risk of tumor recurrence. Traditional single-target CAR-T cell therapy may result in unsatisfactory efficacy when treating certain types of glioblastoma due to tumor heterogeneity and antigen escape mechanisms. However, through the dual-target strategy of the present invention, CAR-T cells can simultaneously recognize and bind two different tumor-associated antigens, thereby reducing the possibility of tumor cell escape and improving the treatment effect. By simultaneously targeting interleukin-13 receptor alpha 2 and NKG2D ligands, the anti-tumor activity of CAR-T cells is significantly enhanced. The dual-target CAR-T strategy provides a novel and effective means for the treatment of glioblastoma and has broad clinical application potential. The dual-target strategy may provide new ideas in improving treatment effect and reducing off-target toxicity.

The present application relates to an index combination, method and system for predicting obesity-related metabolic syndrome, which comprises an index combination for predicting obesity-related metabolic syndrome, the index combination comprising the following indexes: NKG2C + Percentage of mitochondrial low membrane potential in NK cells (NKG2C + NK.MMP low Percentage), NKG2D + Percentage of mitochondrial low membrane potential in NKT cells (NKG2D + NKT.MMP low Percentage), NKG2C + Percentage of mitochondrial low membrane potential in NKT cells (NKG2C + NKT.MMP low Percentage), percentage of mitochondrial low membrane potential in monocytes (Mono.MMP low Percentage), NKG2A + Mitochondrial mass in lymphocytes (NKG2A + Lymph.MM), and mean platelet volume (MPV). The index combination obtained by screening has high biological interpretability and clinical relevance, and can effectively reflect obesity-related immune metabolic abnormalities.

The present invention is directed to a novel targeted heterodimeric Fc fusion proteins comprising an IL-15 / IL-15Rα Fc fusion protein and an NKG2D antigen binding domain Fc fusion proteins.

The invention provides an anti-MIC humanized single-chain antibody as well as a coding gene, an expression vector, a host, a conjugate or conjugate, an adsorbent and application thereof, and the humanized single-chain antibody provided by the invention can be specifically combined with an NKG2D ligand in human blood or plasma so as to relieve a tumor immunosuppression effect caused by abnormal increase of soluble MIC protein. Compared with the traditional antibody, the humanized single-chain antibody provided by the invention has the advantages of small molecular weight, strong stability, low immunogenicity, low cost, easy expression and high expression quantity, and is suitable for large-scale production. According to the invention, the humanized single-chain antibody is combined with the solid-phase carrier, so that the adsorption efficiency is high, the adsorption performance is stable, the non-specific adsorption capacity on other proteins in plasma is low, the specificity is good, the safety performance in clinical treatment is good, and the economic benefit is high. In addition, the humanized single-chain antibody provided by the invention reduces side effects caused by falling of adsorption ligands, and improves safety.

The invention relates to an index combination, a method and a system for predicting obesity-related metabolic syndrome, which comprise an index combination for predicting obesity-related metabolic syndrome. The index combination comprises the following indexes: the percentage of mitochondrial low membrane potential in NKG2C + NK cells (NKG2C + NK.MMPlow%), the percentage of mitochondrial low membrane potential in NKG2D + NKT cells (NKG2D + NKT.MMPlow%), the percentage of mitochondrial low membrane potential in NKG2C + NKT cells (NKG2C + NKT.MMPlow%), the percentage of mitochondrial low membrane potential in mononuclear cells (Mono.MMPlow%), the percentage of mitochondrial mass in NKG2A + lymphocyte (NKG2A + Lymp.MM) and the average platelet volume (MPV). The index combination obtained by screening has relatively high biological interpretation and clinical correlation, and can effectively reflect obesity-related immunometabolism abnormality.

This invention provides a method for screening peptides that specifically target T cells, comprising providing T cells having a subset selected from CD1, CD2, CD3, CD4, CD5, CD7, CD8, CD16, CD25, CD26, CD27, CD28, CD30, CD38, CD39, CD40L, CD44, CD45, CD62L, CD69, CD73, CD80, CD83, CD86, CD95, CD103, CD119, CD126, CD150, CD152 (CTLA-4), CD153, CD154 (CD40L) The T cells are labeled with at least one of the following surface antigen markers: CD161, CD183, CD223, CD254, CD275, CD45RA, CXCR3, CXCR5, FasL, IL18R1, CTLA-4, OX40, GITR, LAG3, ICOS, PD-1, leu-12, TCR, TLR1, TLR2, TLR3, TLR4, TLR6, NKG2D, CCR, CCR1, CCR2, CCR4, CCR6, and CCR7. The T cells are then contacted with a peptide display library, and target peptides that specifically bind to the T cells are screened therefrom.

The invention provides an application of in-vitro amplified NK cells in treatment of autoimmune diseases, the NK cells are obtained by performing in-vitro induced amplification on peripheral blood mononuclear cells, and the in-vitro induced amplification is performed in a culture medium containing interleukin and / or an antibody. Therefore, the NK cell disclosed by the invention has double advantages that on one hand, the NK cell shows phenotypic characteristics of a CD56light tissue resident type NK cell, has the capability of penetrating pathological tissues, and can be used for directly removing abnormal B cells and T cells in tissues of patients with autoimmune diseases; on the other hand, activated receptors such as NKG2D and CD16 and chemokine receptors such as CXCR3 can be stably and highly expressed, and the removal efficiency is further improved by enhancing the recognition and killing capacity on target cells.

The present invention relates to a pharmaceutical composition for preventing or treating cancer, the pharmaceutical composition containing CD4+ CTL as an active ingredient. The CD4+ CTL according to the present invention performs a cytotoxic function of directly attacking and killing myeloma cells, thereby extending the progression-free survival of transplant-ineligible myeloma patients, and thus can contribute to increasing the survival rate of the patients. In addition, by using NKG2D+ CD4+ CTL as a biomarker for predicting the prognosis of myeloma patients, it is possible to analyze the CD4+ CTL status of individual patients and thereby establish customized treatment strategies. In addition, by increasing the cytotoxicity of NKG2D+ CD4+ CTL through a specific cytokine treatment, it is possible to more effectively prevent or treat cancer.

Provided is a method for screening polypeptides that specifically target T cells, comprising: providing T cells having a surface antigen marker selected from at least one of CD1, CD2, CD3, CD4, CD5, CD7, CD8, CD16, CD25, CD26, CD27, CD28, CD30, CD38, CD39, CD40L, CD44, CD45, CD62L, CD69, CD73, CD80, CD83, CD86, CD95, CD103, CD119, CD126, CD150, CD152 (CTLA-4), CD153, CD154 (CD40L), CD161, CD183, CD223, CD254, CD275, CD45RA, CXCR3, CXCR5, FasL, IL18R1, CTLA-4, OX40, GITR, LAG3, ICOS, PD-1, leu-12, TCR, TLR1, TLR2, TLR3, TLR4, TLR6, NKG2D, CCR, CCR1, CCR2, CCR4, CCR6, and CCR7; and contacting the T cells with a polypeptide display library, and screening therefrom target polypeptides that specifically bind to the T cells.