135 results about "NKG2D" patented technology

The invention relates to isolated fully human monoclonal antibodies having specificity for human NKG2D and compositions thereof. The invention further relates to methods for using such antibodies in treating diseases or conditions such as cancer, autoimmune disease, or infectious disease.

The invention relates to isolated fully human monoclonal antibodies having specificity for human NKG2D and compositions thereof. The invention further relates to methods for using such antibodies in treating diseases or conditions such as cancer, autoimmune disease, or infectious disease.

The present invention provides prognostic and diagnostic methods for cancer, as well as methods for monitoring or staging cancer. Methods involve assaying for tumor-derived soluble MIC polypeptide—either MICA or MICB or both—in a sample from a subject. Assays can be implemented with a MIC polypeptide binding agent such as a MIC polypeptide antibody or recombinant NKG2D. An ELISA sandwich assay is employed in some embodiments of the invention to identify a soluble MIC polypeptide. In additional embodiments, a sample is assayed for tumor cell-surface bound MIC in addition to assaying for soluble MIC. The invention also provides methods of cancer therapy involving detecting cancer in the subject by assaying for soluble MIC polypeptide and then administering a cancer therapy.

The invention provides a kit and a method for comprehensively evaluating functions of immune cells in human peripheral blood. The kit for comprehensively evaluating the functions of the immune cells in the human peripheral blood comprises an anti-human CD3, CD4, CD8, CD19, CD21, CD24, CD25, CD27, CD28, CD38, CD56, CD57, CD94, CD127, CD45RA, CXCR3, CXCR5, CCR4, CCR6, CCR7, HLA-DR, PD-1, P30, P46, NKG2D, KIR (NKB1), gamma delta, v delta 2, IgD and IgM antibodies, wherein the antibodies carry fluorescein labels. The kit can be used for comprehensively evaluating the functions of the immune cellsin the human peripheral blood, and is convenient and safe to use.

The invention provides an NKG2D-CAR-T (Chimeric Antigen Receptor-T) cell of a co-expression cell factor IL-7 and application thereof to tumor killing, and particularly provides a CAR of a specific targeted NKG2D ligand protein and a CAR-T cell thereof. The CAR comprises an antigen binding domain and / or IL-7. The CAR-T cell provided by the invention has a good tumor killing effect.

The invention belongs to the technical field of genetically engineered antibody, in particular relates to a fusion protein of tumor vascular endothelial growth factor receptor 2 (VEGFR2 / KDR3) antibody and MICA as well as a preparation method and use thereof. In the invention, a full-length antibody of VEGFR2 is linked with a ligand MHC I molecule associated protein A (MICA) of activated receptor of NK cell (NKG2D) through a flexible linker and expressed by CHO cells by utilizing the genetic engineering technology, and the formed fusion protein can act on VEGFR on the surface of a tumor cell to inhibit or kill tumor and inhibit or destroy tumor angiogenesis, and on the other hand, the formed fusion protein can increase the level of MICA on the surface of the tumor cell and stimulate the NK cells to kill tumor cells through identification of NKG2D on the surfaces of the NK cells, thereby reestablishing the immunological surveillance of an body activated by the NKG2D pathway, and enhancing effects of antibody-dependent cell-mediated cytotoxicity (ADCC) and the like mediated by the Fc region of the antibody to kill tumor cells.

The invention provides a recombinant lentiviral expression vector, and belongs to the technical field of cell engineering. The recombinant lentiviral expression vector comprises a 21mb gene, a 41BBL gene and an MICA gene which are sequentially connected in series; and the 21mb gene is obtained by sequentially connecting a signal peptide gene, an IL21 gene, a CD8 Hinge gene and a CD8 transmembranegene in series. After the provided recombinant lentiviral expression vector transfects a host cell, the host cell expresses the MICA and the 41BBL, and the IL21 is fused and expressed (but not secreted in a culture environment) on a cytomembrane of the host cell. After the provided recombinant lentiviral expression vector transfects the host cell, an obtained reconstitution cell promotes the highexpression of the NKG2D, avoids excessive activation of an NK cell and prolongs the proliferation time of the NK cell in the process of induction culture of the NK cell.

The invention relates to a double-specific chimeric antigen receptor T cell as well as a preparation method and application thereof. The double-specific chimeric antigen receptor double-specifically targeting human NKG2DL and human CD47 is constructed on the basis of human NKG2D and human SIRPalpha molecules as well as an immune response cell modified with the double-specific chimeric antigen receptor. The novel modified immune response cell can effectively realize targeting attack of multiple tumor cells, particularly positive tumor cells expressing NKG2DL and CD47 and can be used for preparing preparations for treating tumor. The preparation method of the immune response cell modified with the double-specific chimeric antigen receptor double-specifically targeting NKG2DL and CD47 comprises simple steps, the obtained immune response cell modified with the double-specific chimeric antigen receptor double-specifically targeting NKG2DL and CD47 has high specific killing rate for tumor cells.

The present invention provides isolated anti-human NKG2D monoclonal antibodies useful for therapeutic applications in humans. Typically, the antibodies are fully human or humanized to minimize the risk for immune responses against the antibodies when administered to a patient. Preferred antibodies include human monoclonal antibodies MS and 21F2. As described herein, other antigen-binding molecules such as, e.g., antigen-binding antibody fragments, antibody derivatives, and multi-specific molecules, can be designed or derived from such antibodies.

A method of decreasing cytokine production and release is disclosed. In one embodiment, the method comprises the step of providing cytotoxic cells to a subject wherein the cells are preferably natural killer cells or T lymphocytes and are genetically modified to express a chimeric antigen receptor comprising a first element that is an extracellular antigen receptor and a second intracellular element that is a signaling moiety comprising altered ADAP-dependent or Fyn-dependent signaling such that downstream signaling causing cytokine release is decreased. Specific modifications of CD137 and NKG2D cytoplasmic tails are described. Additionally, methods to develop and screen drug compounds capable of compromising the binding between ADAP and Fyn and disrupting the downstream release of cytokines are described.

The invention provides a chimeric antigen receptor, a NKG2D CAR-NK cell expressing the chimeric antigen receptor, a preparation method and an application thereof. The chimeric antigen receptor comprises an antigen binding domain, a transmembrane domain, and a costimulatory signaling domain, and the antigen binding domain is capable of specifically binding to a tumor specific antigen NKG2D ligand and activating NK cells through the transmembrane domain and the costimulatory signaling domain. The CAR-NK cell uses the NKG2D ligand as a target antigen and specifically uses the NKG2D CAR-NK cells to kill tumor cells. The cell can be used as a therapeutic drug for tumor diseases for the treatment of tumors with high expression of the ligand of NKG2D molecules, and provides a novel method for tumor prevention and treatment.

The invention discloses a double-target chimeric antigen receptor targeting an NKG2D ligand and CD19 as well as an expression vector and an application thereof, which belong to the field of tumor immune drugs. The double-target chimeric antigen receptor comprises a signal peptide, a NKG2D extracellular region of a targeted NKG2D ligand, a connecting fragment, a single-chain antibody of the targeted CD19, a lengthened CD8 alpha hinge region, a transmembrane region, a costimulatory factor and an intracellular signal peptide which are connected in sequence; the double-target chimeric antigen receptor can be used for specifically identifying tumor cells expressed by two single targets, namely a targeted NKG2D ligand or a CD19 antigen; in addition, tumor cells co-expressed by the targeting NKG2D ligand and the CD19 antigen can be recognized, the double-target CAR-T has stronger anti-tumor activity, immune escape of positive tumor cells expressed by low-abundance antigen can be avoided, andthus the recurrence risk is reduced.

The invention provides a kit for assessing a function of cytotoxic immune cells in human peripheral blood and an assessment method. The kit for assessing the function of cytotoxic immune cells in human peripheral blood provided by the invention comprises an anti-human CD3 antibody, an anti-human CD8 antibody, an anti-human CD45RA antibody, an anti-human CCR7 antibody, an anti-human CD28 antibody,an anti-human CD38 antibody, an anti-human CD57 antibody, an anti-human HLA-DR antibody, an anti-human PD-1 antibody, an anti-human CD56 antibody, an anti-human CD94 antibody, an anti-human NKP30 antibody, an anti-human NKP46 antibody, an anti-human NKG2D antibody, an anti-human KIR antibody, an anti-human Gamma Delta antibody and an anti-human V Delta 2 antibody, wherein all the antibodies are labeled by fluorescein. The kit for assessing the function of cytotoxic immune cells in human peripheral blood provided by the invention can be used for comprehensively assessing the immunity function of the cytotoxic immune cells in human peripheral blood, and is convenient and safe for use.

The invention provides preparation and application of CXCL13 cytokine chemotaxis type CAR-T cells, and particularly engineering immunocyte. The engineering immunocyte can express chimeric antigen receptors CAR and CXCR5 targetting MICA. The engineering immunocyte can selectively kill tumor cells, such as tumor cells high in expression of NKG2D ligands or NKG2D ligands and CXCL13 at the same time,and besides, the CAR-T cells disclosed by the invention have notable killing effects on the tumor cells.

The invention provides a novel chimeric receptor composition, a recombinant vector, a cell and application thereof. The novel chimeric receptor composition comprises a traditional chimeric antigen receptor and an NKG2D chimeric receptor comprising a full-length sequence of NKG2D, DAP10 and / or DAP12 or a truncated fragment, called Dual CAR for short. The chimeric receptor composition provided by the invention enables conventional CAR-T cells to express an NKG2D extracellular region and an intracellular signal region, enlarges a CAR-T antigen recognition spectrum, solves the problem of tumour heterogeneity, enhances the killing ability of CAR-T on tumour cells expressing target antigens, realizes lower-level factor release, reduces the possibility of factor storm, and increases the safety of CAR-T.

The invention relates to a nanometer gene vaccine as well as a preparation method and an application thereof, belonging to the technical field of biology. In order to overcome the defects such as high preparation time consumption, complicated steps and high cost existing in recombinant proteins, the invention provides a nanometer gene vaccine. According to the nanometer gene vaccine, based on a genetic engineering technology, a signal peptide sequence, two identical NKG2D extracellular sequences and a sequence of cell factors IL-15 needed by eukaryotic expression are introduced, and an expression vector of fused proteins is constructed and is prepared into the nanometer gene vaccine by using a biocompatible chitosan material. Because the nanometer gene vaccine carries positive charges, the vaccine can be actively devoured by tumor cells or muscle cells, and proteins dsNKG2D-IL-15 linked between the tumor cells and lymphocytes are expressed in the eukaryocyte. Therefore, natural killer cells (NK) or cytotoxic T cells (CTL) in vivo are activated, the specific immune response in vivo can be enhanced, and an antitumor drug is prepared.

The invention provides a chimeric antigen receptor (CAR) targeting NKG2D. Amino acid sequences of the CAR targeting NKG2D comprises the amino acid sequences of a single-chain antibody targeting NKG2D,an extracellular hinge region, a transmembrane region and an intracellular signal region which are sequentially connected from an amino terminal to a carboxyl terminal, respectively. The single-chainantibody targeting NKG2D comprises the amino acid sequences shown in SEQ ID NO: 1. The invention also provides a chimeric antigen receptor T-cell (CAR-T) targeting NKG2D and a preparation method thereof, as well as application of the CAR targeting NKG2D and the CAR-T targeting NKG2D in the prevention, diagnosis and treatment of liver cancer, cervical cancer, pancreatic cancer and other malignanttumors.

The invention discloses a novel anti-tumor NK cell as well as a preparation method and application thereof. The anti-tumor NK cell is an NK cell transformed by a chimeric co-stimulatory transformationreceptor; the chimeric co-stimulatory transformation receptor consists of a PD-1 extracellular fragment, an NKG2D transmembrane and intracellular fragment and a co-simulatory molecular 41BB intracellular fragment. The novel anti-tumor NK cell disclosed by the invention has the beneficial effects that the chimeric co-stimulatory transformation receptor is adopted for transforming the NK cell, so that the anti-tumor treating effect of the NK cell can be improved, the specific recognition and killing for solid tumor are realized, and no extreme immunized response is caused to an organism.

The present invention relates to the technical field of immunology and the treatment of diseases mediated and / or indicated by the presence of a ligand of NK receptor NKG2D, such as major histocompatibility complex class I chain related (MIC) A and B molecules on the cell surface of diseased tissue or cells. In particular, the present invention makes use of the surprising finding that the stress-inducible endogenous danger signals HSP70 and MICA / B synergistically activate NK cells against tumors. Methods and compositions for the prevention and treatment of infectious diseases, primary and in particular metastatic neoplastic diseases, including, but not limited to human sarcomas, carcinomas and melanomas are provided.

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 belongs to the technical field of genetic engineering antibodies, and particularly relates to a preparation method and application of CD24 antibody fusion protein. The preparation method includes that genetic engineering technology is utilized to connect CD24 single-strand antibody rG7S with MICA extracellular 1-3 zones through G4S flexible peptide, and a mammal eukaryotic expression system stably expresses rG7S-MICA; the CD24 antibody fusion protein rG7S-MICA can be combined with CD24 molecules on the surface of tumor cells and an activated receptor NKG2D on the surface of NK cells at the same time, and can remodel immunological surveillance effect, on CD24+tumor cells, of NK cells induced through an NKG2D path to finally achieve the objective of activating a body autoimmune system to effectively kill the CD24+tumor cells.

The invention belongs to the technical field of biology, and relates to a method for differentiating human induced pluripotent stem cells into natural killer cells. The method comprises the following steps of 1, differentiating the human induced pluripotent stem cells to form endothelial progenitor cells with expression of KDR+CD73+; 2, performing endothelial-blood conversion on the endothelial progenitor cells with the expression of KDR+CD73+ to form NK progenitor cells with the expression of CD34-CD45+; and 3, further differentiating the NK progenitor cells into NK cells with expression of CD56+CD3-. The NK cells produced by the scheme provided by the invention highly express NKG2D receptors, and have stronger cytokine secretion ability and tumor killing ability.

The invention discloses a construction method and application of an NKG2D-ACE2 CAR-NK cell secreting super IL 15. An expression vector mainly comprises an NKG2D extracellular segment, an ACE2 extracellular segment, 4-1BB, CD3[zeta] and IL15R[alpha]-IL15 thawing protein. NKG2D is an activation receptor of the NK cell and is used for identifying ligands (MICA, MICB, ULBP1-6) on the surfaces of virusinfected cells or tumor cells; ACE2 is a receptor of SARS-CoV-2 and is combined with S protein of a virus envelope; the NK cell modified by CAR is proved to be capable of breaking through the limitation of an inhibitory receptor to activate the NK cell, so that the specific killing effect of the NK cell on target cells is enhanced, and no serious toxic or side effect exists; and the activity of an IL15 super agonist is increased by 20 times compared with that of natural IL15 in the body, and the pharmacokinetic property is improved, so that the CAR-NK cell has enhanced durability and target cell killing activity. Therefore, the ACE2 and the NKG2D respectively target the S protein of the SARS-CoV-2 virus and NKG2DL on the surfaces of the infected cells, and the powerful synergistic effectof the super IL15 is achieved, so that the NKG2D-ACE2 CAR-NK cell can specifically remove SARS-CoV-2 virus particles and the infected cells, and a safe and effective cell therapy is provided for COVID-19.

The invention provides an immune cell function evaluation kit and evaluation method for a tumor patient. The immune cell function evaluation kit for the tumor patient comprises an anti-human CD3 antibody, an anti-human CD4 antibody, an anti-human CD8 antibody, an anti-human CD25 antibody, an anti-human CD28 antibody, an anti-human CD38 antibody, an anti-human CD56 antibody, an anti-human CD57 antibody, an anti-human CD94 antibody, an anti-human CD127 antibody, an anti-human CD45RA antibody, an anti-human CCR7 antibody, an anti-human HLA-DR antibody, an anti-human PD-1 antibody, an anti-human NKP30 antibody, an anti-human NKP46 antibody, an anti-human NKG2D antibody, an anti-human KIR antibody, an anti-human gamma-sigma antibody and an anti-human Vsigma2 antibody. All the above antibodies carry fluorochrome labels. The kit can be used for comprehensively evaluating an immune cell function of the tumor patient and is convenient and safe to use.

The invention discloses construction and application of a third-generation NKG2D chimeric antigen receptor T or NK cell. The third-generation NKG2D chimeric antigen receptor T or NK cell mainly comprises a total length NKG2D,41BB costimulatory molecules, and CD3z ; according to the construction and application, the structure uses both 41BB co-stimulatory molecules and NKG2D natural co-stimulatorymolecules DAP10, and NKG2D is an NK cell surface-activated receptor, and can bind to eight ligands (MICA, MICB, ULBP1-6) of NKG2D; the NKG2D ligand (NKG2DL) is not expressed on the surface of a normalcell or the expression quantity is very small, but when the cell is infected by pathogens or cancerated or aged, the expression quantity of the ligands can be greatly increased, so that the ligands become ideal immunotherapy targets.

The invention discloses a bispecific chimeric antigen receptor targeting CD123 and NKG2D ligands and the application of the bispecific chimeric antigen receptor, and particularly discloses a bispecific chimeric antigen receptor (CAR) amino acid construct or a functional variant thereof, wherein the bispecific chimeric antigen receptor (CAR) amino acid construct can simultaneously expresses an anti-CD123 single-chain antibody and a natural NKG2D extracellular fragment and can target CD123 and NKG2DL; the bispecific chimeric antigen receptor has a parallel connection mode and a series connectionmode; and the invention discloses a CAR (chimeric antigen receptor), a construct structure thereof, a nucleotide sequence of the construct, a recombinant expression vector containing the nucleotide sequence and a construction mode and application of the corresponding expression vector. The CAR structure endows T cells with higher and lasting multiplication capacity and high anti-tumor capacity; and according to the bispecific chimeric antigen receptor, multiple infusions of single-target CAR-T cells can be avoided, so that not only is the harm to a patient reduced, but also the economic pressure of the patient can be reduced, and the bispecific chimeric antigen receptor has relatively great clinical research and application values.

The invention belongs to the field of chimeric antigen receptor cells, and particularly relates to a coding amino acid sequence of a bispecific chimeric antigen receptor targeting human CLTX-R and NKG2DL, an immune response cell modified with the coding amino acid sequence as well as a preparation method and application of the immune response cell. The invention relates to a bispecific chimeric antigen receptor of a bispecific targeting human NKG2DL and chlorotoxin receptor (CLTX-R) protein or a functional variant thereof, an immune response cell modified with the bispecific chimeric antigen receptor, as well as a preparation method and application of the bispecific chimeric antigen receptor. The obtained novel functionalized immune response cell can effectively target and attack various tumors, and can be used for preparing a preparation for treating malignant tumors. The immune response cell modified with the bispecific chimeric antigen receptor targeting CLTX-R and NKG2DL can enhance the binding with tumor cells, and obviously enhances the anti-tumor activity.

The invention provides an enhanced immune cell and an application thereof. The immune cell expresses a chimeric antigen receptor, the chimeric antigen receptor comprises an antigen binding structuraldomain, a transmembrane structural domain and a signal transduction structural domain, the antigen binding structural domain comprises an anti-GPC3 single-chain antibody and NKG2D, the chimeric antigen receptor disclosed by the invention has a targeting effect on both GPC3 specific tumor cells and heterogeneous tumor cells, and the constructed immune cell for expressing the chimeric antigen receptor mobilizes the function of endogenous NKG2D while targeting tumor cells for expressing specific antigens, targets tumor cells with heterogeneity, and realizes the clearing and killing effects on specific tumor cells and heterogeneity tumor cells.