57 results about "Granzyme B" patented technology

The invention discloses a preparation method and application of a granzyme B response near-infrared two-region ratiometric fluorescent probe (DCGA). The DCGA combines the NIR-II dual-emission fluorescence characteristic of rare earth ions with a ratio-type signal response mode, the sensitivity and signal stability of in-vivo fluorescence imaging are greatly improved, ultralow or enhanced F1060nm fluorescence signals are respectively shown before and after interaction with granzyme B (GzmB), F1525nm fluorescence signals are almost kept unchanged, and the DCGA can be used for detecting the in-vivo fluorescence imaging of the granzyme B (GzmB). Therefore, the self-calibration ratio fluorescence imaging GzmB activity change is realized. The DCGA is applied to tumor treatment of immunotherapy drugs, and responders and non-responders of immunotherapy can be distinguished in a non-invasive early stage through ratio fluorescence signals of the DCGA. Therefore, the fluorescent probe can be used as an effective GzmB active NIR-II ratio fluorescence imaging tool, and is expected to provide powerful support for immunotherapy research, precision medicine and personalized diagnosis and treatment.

The invention relates to a kit for detecting granzyme B as well as a preparation method and application of the kit. The kit for detecting granzyme B comprises a magnetic microsphere working solution, a detection antibody and streptavidin labeled phycoerythrin. The invention also provides a preparation method of the kit for detecting granzyme B. The preparation method comprises the following steps: coupling a granzyme B antibody by using a magnetic fluorescence coding microsphere to obtain a granzyme B antibody-coupled magnetic fluorescence coding microsphere; labeling the granzyme B detection antibody with biotin to obtain a detection antibody; the preparation method comprises the following steps: mixing granzyme B antibody coupled magnetic fluorescence coded microspheres, a PBS buffer solution, a stabilizer, a preservative, a surfactant and a blocking agent to obtain a magnetic microsphere working solution, and assembling the magnetic microsphere working solution with a detection antibody and streptavidin labeled phycoerythrin to obtain the kit. The invention also provides a kit for detecting serum granzyme B. The invention solves the problem of poor detection reliability caused by false positive results and cross reaction in the existing serum granzyme B detection method.

The invention relates to an antibody composition for detecting abnormal CTL (cytotoxic T lymphocyte) cells of hemophagocytic syndrome and application of the antibody composition, and the antibody composition comprises a CD2 antibody, a CD3 antibody, a CD4 antibody, a CD5 antibody, a CD7 antibody, a CD8 antibody, a CD56 antibody, a CD57 antibody, a Granzyme B antibody, a Perforin antibody, a TRBC1 antibody and a CD45 antibody. In the technical scheme provided by the invention, key phenotypic characteristics of CTL (Cytotoxic Lymphocyte) can be completely covered by various antibodies aiming at T cell and NK cell surface markers, T large granular lymphocyte markers, cytotoxic granular proteins, T cell receptor constant regions and leukocyte common antigens; wherein the clonality of the CTL cells is judged through the expression level of the TRBC1 antibody, and the cytotoxic function activity of the CTL cells is evaluated through the expression levels of Granzyme B and Perforin.

Abstract Described herein, in one aspect, are antigen presenting cells (APCs) comprising an exogenous nucleic acid encoding one or more candidate antigens, wherein the one or more candidate antigens are expressed and presented with MHC class I or MC class II molecules; a molecular reporter of Granzyme B (GzB) activity; and c) an exogenous inhibitor of caspase-activated deoxyribonuclease (CAD)-mediated DNA degradation, a CAD knockout, or a caspase knockout (e.g., caspase 3 knockout). Described herein, in another aspect, is a system for detection of recognized antigen presentation by an antigen presenting cell to a cytotoxic lymphocyte or NK cell. Abstract 2018 / 22761 oM - cell Target ml Target cell cell cell Target Target Target SUBSTITUTE SHEET (RULE 26) cell cell cell Target Target cell cell 1 / 28 my Isolate recognized cell Library of target cells target cells and displaying different Add T cells from sample sequence antigens antigens of interest. CTLs deliver cytotoxic granules to target cells displaying cognate antigen FIG. 1 PCT / US2018 / 036663 20 26 20 53 56 07 J ul 2 02 6 2 0 2 6 2 0 5 3 5 6 0 7 J u l 2 0 2 6 2 0 1 8 / 2 2 7 6 1 o M a n d m y 1 / 2 8 m y L i b r a r y o f t a r g e t c e l l s d i s p l a y i n g d i f f e r e n tA d d T c e l l s f r o m s a m p l e of interest. CTLs deliver c y t o t o x i c g r a n u l e s t o t a r g e t c e l l s d i s p l a y i n g c o g n a t e a n t i g e n P C T / U S 2 0 1 8 / 0 3 6 6 6 3

The invention relates to a three-dimensional cellular microcompartment or an assembly of three-dimensional cellular microcompartments of ovoid, cylindrical, spheroidal or spherical shape or substantially ovoid, cylindrical, spheroidal or spherical shape, the smallest dimension of which is between 200 and 400 μm, comprising an external hydrogel layer defining an internal part, said internal part having a granzyme B content of less than 1 μg / ml of medium and comprising between 500 and 5000 lymphocytes forming a three-dimensionally grouped culture. The invention also relates to a method for producing such a microcompartment or microcompartment assembly.

The invention discloses shRNA (short hairpin Ribonucleic Acid) targeting Zeb2, a recombinant vector, a cell and application of the shRNA in preparation of drugs for enhancing anti-tumor immune response, and belongs to the technical field of research and development of anti-tumor drugs. Experiments prove that the shRNA targeting Zeb2 can reduce the proportion of T cells (Tex, PD1 + Tim3 +), increase the proportion of depleted precursor T cells (Tpex, Ly108 + Tim3-), improve the secretion ability of Granzyme B and TNF-alpha cytokines of CD8 + T cells in tumor tissues, significantly inhibit the tumor growth of skin melanoma mice, increase the infiltration quantity of CD8 + T lymphocytes in the tumor tissues and the anti-tumor ability of the CD8 + T cells, and can be used for preparing anti-tumor drugs for treating skin melanoma. The polypeptide has the potential of being used for preparing drugs for enhancing anti-tumor immune response.

The invention discloses application of an ATP (adenosine triphosphate) citrate lyase (ACLY) inhibitor in preparation of a medicine for treating leucoderma, and belongs to the technical field of preparation of medicines for treating leucoderma. Through targeted inhibition of ACLY activity or specific knockout of ACLY in CD8 + T cells, abnormal activation of ACLY and secretion of killing effector molecules (Granzyme B, IFN-gamma, Perforin and the like) are inhibited, and infiltration of the CD8 + T cells to skin lesion parts is reduced, so that melanocytes are protected from immune injury, and the progress of vitiligo is blocked. The application is based on the abnormally high expression of ACLY in CD8 + T cells of vitiligo patients and the close association with disease activity, and a brand new targeted treatment thought and a medicine research and development direction are provided for vitiligo.

The invention relates to the technical field of biological medicine, in particular to an immune synergist, an enhanced NK cell preparation, a preparation method and application. The immune synergist is hirudin, and the concentration of the hirudin is 10-40 [mu] g / ml. The enhanced NK cell preparation is prepared by the preparation method. According to the preparation method, the enhanced NK cell preparation is obtained after NK cells which are amplified in vitro for 14 days are pretreated by adopting the immune synergist. The application comprises an application of the immune synergist in up-regulation of expression of NK cell granzyme B and an application of the immune synergist in preparation of an NK cell preparation for inhibiting breast cancer cell activity. The expression level of granzyme B of the NK cells can be safely and economically improved, and the in-vitro killing activity of the NK cells to breast cancer cells is remarkably enhanced.

A seranostic agent targeting granzyme B for use in diagnosing and treating cancer. Exemplary therapeutic compounds targeting granzyme B can have the structure of formula (I) disclosed herein. Also provided herein is a method for treating cancer, the method comprising administering to a subject in need of treatment a granzyme B-targeted therapeutic compound disclosed herein.

The invention relates to the technical field of biological medicine, in particular to a TIM-3 small molecule binding agent, a screening method and application, the TIM-3 small molecule binding agent comprises a small molecule compound bound with TIM-3 protein, and compared with an antibody drug, the small molecule compound is small in molecular weight, high in penetrating power and capable of easily penetrating through a cell membrane, so that the TIM-3 protein binding agent can be used for binding TIM-3 protein. The secretion of cell factors such as gamma interferon, interleukin-2, tumor necrosis factor-alpha, granzyme B and the like by T cells is effectively enhanced, so that the compound is expected to be applied to the preparation of immunomodulatory preparations; according to the screening method, firstly, compounds in a compound library are preliminarily screened through a molecular docking platform, then the binding capacity of the screened compounds and TIM-3 protein is verified through the surface ion resonance technology, small molecule compounds with pharmaceutical potential are screened out, and the screening method greatly saves the time and cost of drug research and development.

The invention discloses an engineered dendritic cell as well as a construction method and application thereof, and belongs to the technical field of immunology and medicine. The engineered dendritic cell is a DC cell induced and modified by adopting an engineering means, and an exogenous mRNA sequence is introduced into the DC cell, so that the engineered dendritic cell can efficiently express recombinant IL-18, IL-15Ralpha and TAA. Meanwhile, an exogenous cell factor IL-15 is added into a DC cell culture system after electrotransfection, so that the survival rate and the maturity degree of the cultured engineered DC cells are improved. The IL-15R alpha expressed by the engineered DC and exogenous IL-15 form a membrane anchoring IL-15 / IL-15R alpha compound, the membrane anchoring IL-15 / IL-15R alpha compound and secreted IL-18 play a synergistic effect, T cell activation and proliferation are effectively promoted, meanwhile, generation of cell factors IFN-gamma and release of cytotoxic molecule granzyme B and perforin are expanded, the specific cytotoxicity of T cells to tumors is improved, and the T cells can be used for treating tumor tumors. And the anti-tumor immune effect is sustained and enhanced.

The invention provides a biomarker for diagnosing GVHD and application of the biomarker. Experiments prove that Ki67 positive proportion, Granzyme B positive proportion and TEM cell proportion in donor-derived T cells can be used as biomarkers for distinguishing GVHD patients and non-GVHD patients, and when donor-derived T cells cannot be distinguished, Ki67 positive proportion, Granzyme B positive proportion and TEM cell proportion in T cells can still be used as biomarkers for distinguishing GVHD patients and non-GVHD patients. The method for diagnosing GVHD by using the biomarker provided by the invention has the advantages of immune subgroup specific quantification capability, low cell quantity adaptability, short timeliness, low cost and the like.

Cyclic peptides that bind to granzyme B and are useful for imaging granzyme B, salts thereof, pharmaceutical compositions comprising the same, diagnostic and therapeutic uses, and methods of making such compounds are disclosed. Compounds useful as radiotracers for positron emission tomography (PET) and / or single photon emission tomography (SPECT) imaging are also provided. Further disclosed is the use of the compounds as imaging agents for granzyme B.

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.

The invention relates to a molecular marker for immunotherapy prediction of a targeting shower cell and application of the molecular marker, and belongs to the technical field of molecular biology. The invention provides a molecular marker for immunotherapy prediction of a targeting shower cell. The molecular marker comprises CD8, CD4, PD-1, Foxp3, CD19 and CD3, wherein the CD8, the CD4, the PD-1, the Foxp3, the CD19 and the CD3 are used for immunotherapy prediction; or comprises a CD45, a CD45RA, a CD45RO, a CD25, a CCR7 and a CD127; or comprises CD3, LAG3, Tim3, ICOS, CD69 and CD103; or comprises a CD79A, a CD79B, a CD20, a CD21 and a CD138; or the CD56, the CD161, the CD57, the CD94 and the Granzyme B are included; the invention relates to a CD56, CD161, CD57, CD94 and Granzyme B. The CD56, the CD161, the CD57, the CD94 and the Granzyme B. The molecular marker can accurately distinguish shower cell groups and interaction.

The invention relates to: a composition for culturing natural killer cells, comprising, as an active ingredient, a fusion protein comprising an IL-2 protein and a CD80 protein; and a method for preparing natural killer cells by using same. Particularly, a composition for culturing natural killer cells, comprising, as an active ingredient, a fusion protein comprising IL-2 or a variant thereof and CD80 or a fragment thereof, of the present invention; promotes the proliferation of natural killer cells, induces the expression of CD16 and NKp46, and increases the expression and secretion of granzyme B and perforin, thereby being effectively usable in the preparation of natural killer cells having an excellent anticancer immune function.

To provide compounds useful for imaging granzyme B by using medical imaging including positron tomography.SOLUTION: A method for imaging granzyme B in a cell or a tissue includes: i) a step of bringing a sample of the cell or tissue in contact with a compound of formula I or a pharmaceutically acceptable salt thereof; and ii) a step of imaging the cell or tissue by an appropriate imaging method and thus imaging granzyme B in the cell or tissue. In the formula, A includes one or a plurality of imaging agents, B denotes an arbitrary selected linking group, and C denotes a group binding to granzyme B.SELECTED DRAWING: None

The application belongs to the technical field of biological medical materials, and particularly relates to a granzyme B specific response nano probe and a preparation method and application thereof. The granzyme B specific response nano probe comprises a near-infrared down-conversion nano probe, and a pentapeptide IEFDK-fluorescence quenching modification group is modified on the surface of the near-infrared down-conversion nano probe; the pentapeptide IEFDK-fluorescence quenching modification group is derived from a pentapeptide IEFDK-fluorescence quencher with a structure shown in formula (1). The granzyme B specific response nano probe provided by the application can specifically respond to granzyme B, realize immune effect monitoring of a tumor site, and thus realize detection of tumor immune responses after different radiotherapy schemes are processed. Formula (1).

The invention belongs to the technical field of biological medicine, and relates to a dendritic cell vaccine as well as a preparation method and application thereof, in particular to a dendritic cell vaccine based on enhancement of dendritic cell maturation and vaccine efficiency and application of the dendritic cell vaccine in tumor treatment. The dendritic cell disclosed by the invention is a dendritic cell with deletion of a metabotropic glutamate receptor 4 or a dendritic cell treated by forskolin, and the dendritic cell can promote the expression of co-stimulatory molecules on the surface of the dendritic cell; the proliferation of T cells is promoted; inducing the effector T cells to secrete interferon gamma and granzyme B; the killing capability of T cells on tumor cells is enhanced. The dendritic cell vaccine disclosed by the invention comprises dendritic cells loaded with antigens, wherein the antigens are liver cancer cell lysates. The dendritic cell vaccine disclosed by the invention has an anti-tumor effect and can be used for preparing anti-tumor medicines.

The present application provides a dendrobium officinale polysaccharide extract, a preparation method and an application, the dendrobium officinale polysaccharide extract is with dendrobium officinale stem as raw material, is extracted by purified water reflux, is precipitated by ethanol, is deproteinized by Sevag, is decolorized and is deashed and is obtained by gel / film separation and purification, the mass percentage content of polysaccharide in the extract is 50-95wt%, the average molecular weight is 10-300 kDa, and the monosaccharide component includes mannose, glucose, galactose, fucose, rhamnose and arabinose.The dendrobium officinale polysaccharide extract provided by the present application has immune activation effect on natural killer cells, can promote the NK cell CD107a degranulation reaction and the expression of IFN-gamma, perforin and granzyme B in a dose-dependent manner, up-regulates the NKG2D related pathway, thereby activating the NK cell function and enhancing the killing activity of NK cells on tumor target cells such as K562.

Compounds of Formula (I) and Formula (II), which are capable of binding to granzyme B. Also provided herein are pharmaceutical compositions comprising such for use in, for example, imaging Granzyme B and / or treating immunoregulatory abnormalities.

The invention relates to an application of beta-damascenone (beta-damascenone) in preparation of a medicine for inhibiting CD8 < + > T cell depletion. The invention discovers that beta-damascenone can significantly down-regulate the expression of a depletion marker TIM3 of CD8 + T cells and maintain the secretion level of Granzyme B and TNF-alpha, so that the cytotoxic function of the T cells is not damaged while the depletion of the T cells is inhibited. The use includes the use of beta-damascenone as a single active ingredient or in combination with an immune checkpoint inhibitor (such as an anti-PD-1 antibody) for the treatment of cancer, such as melanoma, non-small cell lung cancer or colorectal cancer. The invention further provides an in-vitro method for enhancing the anti-tumor function of the CD8 + T cells.

The invention relates to the technical field of biology, in particular to a TLR2 / TLR4 activating peptide-based CAR-T cell as well as a preparation method and application thereof. Through lentiviral vector transduction, the T cell co-expresses the following components: a) a mesothelin-targeted chimeric antigen receptor CAR; and b) an exogenous oligopeptide EEIYEEFA (SEQ ID NO: 1), which is used for up-regulating downstream NF-kappa B and MAPK signal channels by activating TLR2 / TLR4 so as to further improve the secretion levels of IFN-gamma, IL-2 and GranzymeB (P is less than 0.01). The invention has the advantages that: a T cell TLR2 / TLR4 signal channel is activated by secreting PEP, and M2 type macrophages are polarized into M1 type macrophages, so that the solid tumor treatment effect is obviously enhanced. The cell inhibits tumor growth by 70% in a lung cancer tumor-bearing model, prolongs the lifetime of mice to 45 days, and has important clinical application value.

Compositions and methods are provided for the cell-mediated targeted killing of diseased cells based on the presence of an intracellular antigen, rather than a surface-bound marker. The targeting cells are modified to express a cytotoxic protein that is delivered into a targeted cell, and after delivery is selectively activated by the presence of a cytoplasmic protein of interest. In one embodiment of the invention, the cytotoxic molecule is a Granzyme B (GrB) polypeptide. In the compositions of the invention, GrB is modified to render its cytotoxic enzymatic functions inactive, until the presence of an intracellular antigen unlocks the GrB molecule to enable enzymatic activities.

Novel cyclic peptides that bind to granzyme B and may be suitable for imaging granzyme B, their salts, pharmaceutical compositions containing them, diagnostic and therapeutic uses, and methods for producing such compounds are disclosed. Additionally, compounds useful as radiotracers for positron emission tomography (PET) and / or single photon emission computed tomography (SPECT) imaging are provided. The use of the compounds as imaging agents for granzyme B is further disclosed.

This invention relates to a granzyme B-targeting complex, a radiopharmaceutical, and their imaging applications in tumor immunotherapy, belonging to the field of nuclear medicine diagnostics. The complex is covalently modified, triggering a covalent group proximity effect by recognizing the binding of a peptide to granzyme B, achieving irreversible covalent linkage between the probe and the target protein. This significantly enhances the probe's selective retention at the target site and extends the imaging time window. After radionuclide labeling, the complex can be prepared into a radiopharmaceutical for nuclear medicine imaging; the labeling process is simple and exhibits good stability. This radiopharmaceutical allows for non-invasive, specific, and quantitative monitoring of granzyme B expression levels via PET or SPECT, reflecting the activity status of immune cells in vivo. This invention has significant clinical application value and promising prospects for the early and accurate prediction and dynamic monitoring of tumor immunotherapy efficacy.

The invention provides a dendrobium officinale polysaccharide extract as well as a preparation method and application thereof.The dendrobium officinale polysaccharide extract is prepared from dendrobium officinale stems as a raw material through purified water reflux extraction, ethanol precipitation, Sevag deproteinization, decoloration and deliming and gel / membrane separation and purification, the mass percentage content of polysaccharide in the extract is 50-95 wt%, the average molecular weight is 10-300 kDa, and the content of polysaccharide in the extract is 50-95%. The monosaccharide components comprise mannose, glucose, galactose, fucose, rhamnose and arabinose. The dendrobium officinale polysaccharide extract provided by the invention has an immune activation effect on natural killer cells, can dose-dependently promote the degranulation reaction of NK cells CD107a and the expression of IFN-gamma, perforin and granzyme B, and up-regulate NKG2D related pathways, so that the functions of the NK cells are activated, and the killing activity of the NK cells on tumor target cells such as K562 is enhanced.

The invention discloses an application of Danjie emodin in preparation of a product for inhibiting T cell depletion. It is found through in-vitro screening that the Danjie emodin can significantly inhibit T cell depletion. Mechanism research shows that the Danjie emodin does not influence the secretion of Granzyme B and slightly inhibits the secretion level of a cell factor IFN-gamma; by inhibiting an AKT phosphorylation signal channel, accumulation of protein aggregates in T cells is remarkably reduced, and the cell protein homeostasis is maintained, so that T cell depletion is inhibited. Animal experiments also prove that the T cells treated by the Danjie emodin can significantly inhibit the growth of melanoma after adoptive reinfusion. Therefore, the invention provides the application of the Danjie emodin in preparation of the product for inhibiting T cell depletion, and further provides a new small molecule metabolism intervention strategy for improving the solid tumor immunotherapy effect.

The application provides an application of dodartin A1 in activating NK cells. Through in-vitro experiments, it is proved that dodartin A1 can significantly promote the synthesis and release of IFN-gamma and cytotoxic substances (such as granzyme B and perforin) of NK92 cells, and increase the expression of the surface agonistic receptors NKp46 and NKG2D of the NK92 cells; through in-vivo experiments, it is proved that dodartin A1 can promote the expression of the NK cell activation related substances CD107a and NKp46 to play an anti-hepatoma effect; in conclusion, dodartin A1 has the effect of activating NK cells.