37 results about "Cell killing" patented technology

A method for treating tumors using a combination of an oncolytic virus vaccine and immune cells. The method specifically includes the following step: treating the tumors by using a combination of the immune cells and the oncolytic virus vaccine; the oncolytic virus vaccine includes a recombinant oncolytic virus expressing a tumor antigen and is used for targeting the tumor cells; the immune cells express a chimeric antigen receptor paired with the tumor antigen, and are used for killing or destroying the tumor cells targeted; the recombinant oncolytic virus includes an M protein, G protein, N protein, P protein and L protein subjected to site-directed mutagenesis. The tumor antigen expressed by the oncolytic virus vaccine can guide the immune cells to reach the target tumor tissue center, achieving a curative effect where 1+1 is greater than 2, with a tumor cell killing rate being up to 100% at most.

Provided are the construction, preparation and use of a functionally enhanced universal CAR-DNT cell. Specifically, provided is a chimeric antigen receptor construct. A CAR function-enhancing element and IL-10 are sequentially fused to the C-terminus of a tumor antigen-targeting CAR via a self-cleaving peptide, and the CAR construct is introduced into a DNT cell, thereby obtaining a functionally enhanced universal CAR-DNT cell, namely, an IL10-CD19-CAR-mbIL15-DNT cell. The cell specifically targets CD19, and has a stronger and more sustained cell killing activity and better safety, thus providing a new therapy for CD19-mediated diseases.

The application discloses application of DUSP4 in preparation of a marker for predicting the curative effect of immunotherapy of hepatocellular carcinoma and a sensitizing drug, and belongs to the technical field of biological medicine.The application finds that high expression of dual specificity phosphatase 4 (DUSP4) is significantly related to high response rate and long survival period of an immunological checkpoint blocking therapy for a hepatocellular carcinoma patient.DUSP4 promotes CD8+ T cell and NK cell infiltration and remodels an immune microenvironment by inhibiting a TGF-beta signal path, down-regulating an immunosuppressive factor and up-regulating an antigen presenting molecule and a chemotactic factor.The application provides a kit and a method for detecting the expression level of DUSP4 to predict an immunotherapy response, and a combined drug composition comprising a DUSP4 activator or a TGF-beta inhibitor and an immunological checkpoint inhibitor.Experiments prove that up-regulation of the expression of DUSP4 can significantly enhance T cell killing function, and produces a synergistic anti-tumor effect with an anti-PD-1 antibody.The application provides a new strategy for precise stratified treatment and overcoming immunological drug resistance of hepatocellular carcinoma.

The invention relates to application of a proteasome inhibitor in preparation of a medicine for treating tumors resistant to a tyrosine kinase inhibitor, and in the application, a plurality of strains of liver cancer cells with drug resistance to lenvatinib or regorafenib are constructed by utilizing a plurality of human liver cancer cell lines with different genetic backgrounds; a high-throughput CRISPR (clustered regularly interspaced short palindromic repeats) gene knockout screening technology and a patent medicine gene CRISPR library are utilized, and gene targets of which the gene knockout or inactivation can better kill drug-resistant cells are specifically identified from thousands of patent medicine genes. Wherein the function inactivation of a plurality of genes in the proteasome family shows more obvious cell killing activity in a plurality of drug-resistant cell models compared with non-drug-resistant cells. The invention discovers and verifies that the proteasome inhibitor has a specific killing effect on tyrosine kinase inhibitor drug-resistant liver cancer for the first time.

An anti-tumor pharmaceutical composition based on immune checkpoint blocking and a use thereof. The anti-tumor pharmaceutical composition comprises: paroxetine hydrochloride and a T cell enhancer, the T cell enhancer being at least one of vitamin E and thymosin. By means of a series of in vivo and in vitro experiments, it was found for the first time that cannabidiol and paroxetine hydrochloride can effectively reduce the expression of PD-L1 on a tumor cell membrane, thereby blocking the PD-1 / PD-L1 signaling pathway and enhancing the effect of T cells in killing tumor cells. T cell enhancer added on this basis can further increase the number and activity of T cells and significantly enhance the killing ability of T cells after relief from immunosuppression, greatly improving the anti-tumor effect of a drug. The components cannabidiol, paroxetine hydrochloride, and vitamin E+thymosin of the pharmaceutical composition can have a synergistic effect to improve the anti-tumor effect.

The application discloses an anti-B7H3 full human monoclonal antibody and a derivative product and application thereof, wherein the derivative product comprises a chimeric antigen receptor targeting B7H3, and the chimeric antigen receptor comprises an antigen binding domain, a hinge region, a transmembrane domain, a costimulatory signal domain and an intracellular signal transduction domain, wherein the antigen binding domain is the anti-B7H3 full human monoclonal antibody, and the antibody has an ADCC effect; the chimeric antigen receptor provided by the application has specific targeting effect on B7H3 positive tumor cells, and T cells expressing the chimeric antigen receptor have significant killing effect, and thus have important significance in the tumor treatment field.

This invention belongs to the field of biomedicine and tumor immunotherapy technology, specifically relating to a monoclonal antibody targeting the immune checkpoint molecule CD276 (B7-H3) and its applications. The applicant conducted immunization and screening work around the human CD276 antigen, selecting two representative monoclonal antibody clones, 2B2-1 and 5G4-2, from multiple candidate antibodies. The nucleotide and amino acid sequences of their heavy and light chain variable regions were determined, and their performance in flow cytometry, T-cell killing assays, immunohistochemistry, ELISA, and Biacore affinity assays was systematically evaluated. Experimental results show that the above antibodies can specifically recognize and bind to CD276, efficiently distinguish between CD276-high and low-expressing cells, and enhance the killing activity of T cells against CD276-high expressing tumor cells in an in vitro co-culture system. This provides an excellent antibody backbone for the subsequent construction of diagnostic kits, ADC drugs, or bispecific antibodies.

Provided are a chemokine CXCL12 fusion protein and a use thereof. The chemokine CXCL12 fusion protein comprises the chemokine CXCL12 and an antibody against a tumor-associated antigen. The chemokine CXCL12 and the antibody against the tumor-associated antigen are directly fused or fused by means of a linker. On the one hand, the provided chemokine CXCL12 fusion protein can promote T cell infiltration and improve targeting. On the other hand, T cell killing signals are highly dependent on target cells, and T cells generate immune signals to kill the target cells only upon coming into contact with the target cells under the mediation of the fusion protein, thereby reducing toxic side effects.

An anti-tumor pharmaceutical composition based on immune checkpoint blocking and a use thereof. The anti-tumor pharmaceutical composition comprises: paroxetine hydrochloride and a T cell enhancer, the T cell enhancer being at least one of vitamin E and thymosin. By means of a series of in vivo and in vitro experiments, it was found for the first time that cannabidiol and paroxetine hydrochloride can effectively reduce the expression of PD-L1 on a tumor cell membrane, thereby blocking the PD-1 / PD-L1 signaling pathway and enhancing the effect of T cells in killing tumor cells. T cell enhancer added on this basis can further increase the number and activity of T cells and significantly enhance the killing ability of T cells after relief from immunosuppression, greatly improving the anti-tumor effect of a drug. The components cannabidiol, paroxetine hydrochloride, and vitamin E+thymosin of the pharmaceutical composition can have a synergistic effect to improve the anti-tumor effect.

The invention provides construction, preparation and application of a universal CAR-DNT (chimeric antigen receptor-DNT) cell with enhanced functions. Specifically, the invention provides a chimeric antigen receptor construct, a CAR function enhancing element and IL-10 are sequentially fused at the C tail end of a CAR of a targeted tumor antigen through a self-cleavage peptide, and the CAR construct is introduced into a DNT cell, so that a function-enhanced universal CAR-DNT cell, namely, an IL10-CD19-CAR-mbIL15-DNT cell, is obtained. The IL10-CD19-CAR-mbIL15-DNT cell disclosed by the invention is specifically targeted to CD19, and has stronger and more lasting cell killing activity and better safety, so that a new therapy is provided for diseases mediated by the CD19.

The application discloses a kind of for NK cell synergistic composition and its in-vitro culture method, it is related to cell culture technical field.The composition for the activation of NK cell includes buffer, and only the following final concentration of cytokine combination is contained in buffer: 100-500U / ml IL-2, 20-50ng / ml IL-7, 5-20ng / ml IL-12, 5-20ng / ml IL-15, 10-50ng / ml IL-18 and 10-30ng / ml IL-21.The composition for the activation of NK cell provided by the application is added to culture medium, the expansion fold and purity of peripheral blood autologous NK cell can be improved, and the killing activity of NK cell is enhanced.It is favorable to improve the curative effect of cell immunotherapy to patient.

The invention discloses application of ME1 in enhancing the anti-tumor function of CAR-T cells, and relates to a preparation method of a chimeric antigen receptor plasmid structure of overexpressed malic enzyme 1 for enhancing the killing activity of the CAR-T cells, in particular to the malic enzyme 1 serving as a gene for enhancing the functions of the CAR-T cells. When the compound is combined with a CAR-T cell treatment product for application, the anti-tumor effect can be remarkably improved.

Provided is a dual-target BCMA / CD19 chimeric antigen receptor construct. Membrane-bound interleukin-15 is fused, via a self-cleaving peptide, at the C-terminus of a bispecific CAR targeting BCMA and CD19, and the CAR construct is introduced into a DNT cell, such that a universal dual-target BCMA / CD19-CAR-DNT cell having enhanced function is obtained. The cell simultaneously targets BCMA and CD19, and has cell-killing activity, thereby providing a new treatment for BCMA- and / or CD19-mediated diseases.

This invention belongs to the field of biomedical technology, specifically relating to an anti-tumor nanomaterial, its preparation method, and its application. The anti-tumor nanomaterial is formed by in-situ growth of calcium peroxide on the surface of black phosphorus nanosheets to create hierarchical black phosphorus-calcium peroxide nanosheets, followed by coating the surface of these composite nanosheets with hyaluronic acid. After entering tumor cells, the anti-tumor nanomaterial releases calcium ions and hydrogen peroxide, triggering an abnormal increase in calcium ions and oxidative stress within the cells, thereby causing mitochondrial dysfunction and inducing tumor cell death. Simultaneously, the released calcium ions combine with phosphate ions produced by the degradation of black phosphorus, forming calcium phosphate mineralization deposits within the tumor tissue, inducing tumor biocalcification. Changes in calcium signaling and the oxidative stress process can also promote the infiltration and activation of immune cells, thereby regulating the tumor immune microenvironment and achieving a synergistic effect of cell killing, biocalcification induction, and immune regulation within tumor tissue.

The invention discloses a humanized antibody targeting human CD33 and application thereof, and belongs to the field of biological medicine. The invention provides a monoclonal antibody targeting human CD33, and also provides a composition containing the monoclonal antibody, a multispecific antibody, a chimeric antigen receptor and an antibody conjugate. The monoclonal antibody targeting human CD33 has the characteristics of high affinity and strong killing effect on target cells, and is beneficial to prediction, diagnosis and treatment monitoring of related diseases such as myelogenous leukemia.

The application provides a method for enhancing the expansion and activation of memory-like NK cells, and belongs to the technical field of cell culture. The application provides a method for obtaining strong memory-like NK cells, which comprises using complete culture medium, IL-2, IL-12, IL-15, IL-18, sodium alpha-ketoglutarate and DOT1L inhibitor; the concentration of the sodium alpha-ketoglutarate is 0.1-2 mM, and the concentration of the DOT1L inhibitor is 10-100 nM. The addition of low-concentration sodium alpha-ketoglutarate in the complete culture medium of memory-like NK cells can promote the in-vivo and in-vitro proliferation capacity of the memory-like NK cells. The addition of low-concentration DOT1L inhibitor in the late culture stage can further enhance the killing function of the memory-like NK cells by inhibiting H3K79me2 / 3, etc., so that memory-like NK cells with a larger number and stronger killing capacity are obtained.

The invention provides a chip model for detecting whether candidate drugs can cause cytokine storm, and belongs to the technical field of biomedicine. The chip model comprises a culture plate, the culture plate comprises a plurality of culture units, and each culture unit comprises a culture solution channel and a plurality of cell growth holes communicated with the culture solution channel; the bottom of the cell growth hole is lower than the bottom of the culture solution channel to form a growth hole concave part. According to the chip model, medicine combined immune cell killing can be carried out while 3D cell culture is realized, and whether side reaction is generated or not is detected in subsequent experiments. According to the application, higher-throughput culture and detection can be carried out, the experimental purpose of detecting and evaluating a plurality of anti-tumor drugs on the same chip is achieved, and drug safety evaluation can be carried out through components of a culture solution while drug effect evaluation is carried out.

The invention relates to the field of bioengineering, in particular to a function-synergetic bionic gene editing carrier system as well as a preparation method and application thereof. The invention provides a gene editing expression box. The gene editing expression box comprises a cancer cell specific promoter, a CXCL9 gene, an sgBAG3 gene and an sgHSP70 gene. A bionic lipid carrier with protein adsorption resistance, cancer cell targeting and membrane fusion delivery is constructed, and an integrated gene editing CRISPR system capable of highly expressing CXCL9 protein factors in cancer cells and double editing tumor cell BAG3 and HSP70 genes is entrapped, so that a multifunctional synergetic bionic gene editing carrier system is constructed; according to the invention, in-vivo delivery stability of the CRISPR system, cancer cell targeting accuracy and intracellular release high efficiency are realized, liver cancer cell gene editing efficiency is improved, a T cell killing enhancement effect tumor immune microenvironment is constructed, and effective cancer resistance of the CRISPR system in a tumor environment is realized.

The invention belongs to the technical field of biological medicine, and particularly discloses a construction method of a metabolite screening platform for regulating and controlling a T cell killing function, tumor cells expressing OVA and CD8 + T cells are co-cultured, and the co-culture is combined with a non-targeted metabonomics technology to form the metabolite screening platform for regulating and controlling the T cell killing function. The metabolite screening platform for regulating and controlling the killing function of the T cells is based on an in-vitro killing model which is clear in antigen, high in specificity and good in repeatability; the model and a high-sensitivity metabolite detection technology can be integrated; key metabolites which change remarkably in the tumor killing process of the T cells can be screened systematically without deviation; a metabolic pathway or a specific molecule which can be used as a biomarker for predicting the curative effect and can also be used as an intervention target to enhance the T cell function can be identified.

The invention provides construction, preparation and application of a universal double-target BCMA (bovine serum albumin) / CD19-CAR-DNT (Chimeric Antigen Receptor-DNT) cell with enhanced functions. Specifically, the invention provides a chimeric antigen receptor construct of double-target BCMA / CD19, a membrane binding interleukin 15 (mbIL15) is fused at the C terminal of a bispecific CAR targeting BCMA and CD19 through a self-cleavage peptide, and the CAR construct is introduced into a DNT cell, so that a universal double-target BCMA / CD19-CAR-DNT cell with enhanced functions is obtained. The BCMA / CD19-CAR-DNT cell disclosed by the invention is targeted to BCMA and CD19 at the same time, and has stronger and more lasting cell killing activity, so that a new therapy is provided for BCMA and / or CD19 mediated diseases.

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.

The present invention relates to the identification of key residues within mouse lgG3 antibodies (mAbs) that are responsible for intermolecular cooperativity and their transfer into lgG1 antibodies in order to enhance their functional affinity and direct cell killing.

The invention discloses a fluorescent probe for detecting carboxylesterase 1 and application of the fluorescent probe, and belongs to the technical field of biological medicine. The specific probe substrate can be used for determining the enzyme activity of CES1 in a biological system. The CES1 enzyme activity determination process is as follows: meso-methyl ester hydrolysis reaction of indocyanine green compound derivatives is selected as probe reaction, and the CES1 enzyme activity in various biological samples is determined by quantitatively detecting the generation amount of metabolites in unit time. The probe can be used for detecting high-expression CES1 in tumors and is used for distinguishing tumor areas from normal tissues. A product generated after CES1 metabolism of the probe can generate active oxygen under illumination of 690 nm, has remarkable cell killing capacity, inhibits growth of tumor cells and changes the forms of the tumor cells. The probe is a selective probe for detecting CES1 activity, so that efficient discovery of CES1 high-expression cells and intestinal bacteria can be realized.

The invention discloses a universal HER2 CAR-T cell for expressing HLA-G as well as a preparation method and application of the universal HER2 CAR-T cell, and belongs to the technical field of biological medicines. According to the present invention, a lentivirus transduction technology and a CRISPR / Cas9 technology are adopted to prepare the universal type CAR-T cell (the universal type HER2 CAR-T cell for expressing the HLA-G targeting HER2), wherein the universal type HER2 CAR-T cell is used for expressing the HLA-G targeting HER2; it is verified that the cell can kill HER2 positive non-small cell lung cancer cells, can resist immunological rejection of allogeneic PBMC cells and can also resist killing of NK92 cells, the survival number of the universal CAR-T is increased, premature exhaustion of the universal CAR-T is avoided, and in a word, a technical foundation is laid for killing HER2 positive tumor cells through the universal CAR-T.

The present application provides a kind of dual-specificity antibody targeting CD3 and SLAMF7 and its use.The antibody includes: the first antigen binding region, the first antigen binding region has CD3 binding activity;Second antigen binding region, the second antigen binding region has SLAMF7 binding activity, the first antigen binding region is selected from CD3 single-chain antibody, and the second antigen binding region includes a SLAMF7 heavy chain and a SLAMF7 light chain.The antibody can be combined with SLAMF7 positive tumor cells and T cells simultaneously, effectively mediate T cell killing tumor cells, and can effectively treat cancer.

The application provides a kind of targeting 4-1BB and Nectin-4 dual-specificity antibody and its use.The antibody includes: the first antigen binding region, the first antigen binding region has 4-1BB binding activity;Second antigen binding region, the second antigen binding region has Nectin-4 binding activity, first antigen binding region is selected from 4-1BB single-chain antibody, and second antigen binding region includes a Nectin-4 heavy chain and a Nectin-4 light chain.The antibody can be combined with Nectin-4 positive tumor cells and T cells simultaneously, effectively mediate T cell killing tumor cells, and can effectively treat cancer.

The invention discloses an immune cell killing activity detection method based on fluorescently labeled tumor target cells, and relates to the technical field of detection.The immune cell killing activity detection method comprises the steps that peripheral blood or umbilical cord blood of a healthy volunteer is collected, mononuclear cells are obtained through separation with a human lymphocyte separation medium and induced into immune effector cells, and multiplication culture is conducted; taking the fluorescently-labeled tumor cells as target cells, and co-incubating the immune effector cells and the target cells for a period of time; collecting a cell product after co-incubation is finished, and detecting the number of fluorescently-labeled positive cells in the co-incubation product by using a flow cytometer or a fluorescent cell counter; and calculating the killing rate of the immune effector cells to the target cells based on the detected number of the positive cells. The scheme is simple and convenient to operate, rapid, accurate, low in cost and free of cell damage.

The invention relates to the technical field of antibodies, in particular to a humanized anti-CD3 single-chain antibody, a bispecific antibody and application of the humanized anti-CD3 single-chain antibody and the bispecific antibody. The technical problem to be solved by the invention is to provide an anti-CD3 / tumor associated antigen bispecific antibody which has no mismatch problem and has tumor cell-dependent T cell activation activity. In order to solve the technical problem, the invention firstly provides a humanized anti-CD3 single-chain antibody, and further provides a bispecific antibody of anti-CD3 and tumor-associated antigens on the basis of the humanized anti-CD3 single-chain antibody. The bispecific antibody shows tumor cell dependent T cell activation activity, has relatively weak T cell activation activity under the condition of no tumor cells, can avoid excessive activation of T cells before being combined with the tumor cells, reduces the occurrence risk of cytokine storm, and has the advantage of safety. The anti-CD3 / tumor associated antigen bispecific antibody provided by the invention can mediate T cells to kill tumor cells, and has application potential in preparation of antitumor drugs.

The application provides a kind of targeting CD27 and Nectin-4 dual-specific antibody and its use.The antibody includes: the first antigen binding region, the first antigen binding region has CD27 binding activity;Second antigen binding region, the second antigen binding region has Nectin-4 binding activity, first antigen binding region is selected from CD27 single-chain antibody, and second antigen binding region includes a Nectin-4 heavy chain and a Nectin-4 light chain.The antibody can be combined with Nectin-4 positive tumor cells and T cells simultaneously, effectively mediate T cell killing tumor cells, and can effectively treat cancer.