52 results about "Perforin" patented technology

A nutrient enriched wheat flour with the function of preventing and treating cancers by generating a lot of perforin is proportionally prepared from wheat flour, soybean powder, corn powder and the powders of coix seed, cassia seed, edible lignin starch, tremella and ganoderma.

The invention aims to provide an efficient amplification method of TILs serving as sources of cancerous pleural effusion. The TILs prepared with the method have the advantages of being high in multiplication speed, large in number, high in cell activity and the like. The method is mainly characterized in that by utilizing a cultivation bottle coated by laminin protein and fibronectin, the TILs are cultivated. The adsorption effect of the two kinds of protein can make cells in half-adherence state, therefore, the cells are favorable for being stimulated by various factors, and the two kinds of protein has the multiplication promoting effect on the TILs; by using a resistant OX-40 monoclone antibody, co-stimulatory signals of CD3+CD8+T cell subgroups in the TILs can be stimulated, cell activation is promoted, the expression of cell perforin and granular enzyme is improved, and the killing capacity of the TILs is enhanced. So far, the study report that the laminin protein, the fibronectin and the resistant OX-40 monoclone antibody are combined and used for preparing the TILs has not been given, and the method for increasing the cell number and improving the killing activity by adding the three factors in preparation of the TILs is put forward for the first time.

The present invention relates to retroviral vectors capable of driving the expression of perforin in a cell and a method of expressing recombinant perforin in a cell. The present invention also relates to recombinant perforin polypeptides and nucleic acid molecules derived therefrom and uses thereof. Also encompassed are screening assays employing such recombinant perforin molecules, compounds identified by the screening assays and uses thereof.

The present invention provides novel compounds of the Formula I, pharmaceutical compositions comprising such compounds and methods for using such compounds as agents or drugs for inhibiting perforin activity and for treating a subject at risk of or susceptible to a disease or disorder, or having a disease or disorder associated with undesirable perforin activity.

The invention discloses an application of an NK cell stock solution in preparation of a medicine for treating skin diseases. An NK cell is an important immune cell for an organism, and not only is related to tumor resistance, viral infection resistance and immune regulation, but also takes part in happening of hypersensitive responses and autoimmune diseases in some cases. The NK cell stock solution contains main components of NK cell excreted proteins and cell factors, including MIP-1alpha, MIP-1beta, RANTES, monocyte chemoattractant proteins, interleukin, colony stimulating factors, tumor necrosis factors, interferon and perforin, and these proteins and cell factors can be absorbed by the skin and directly act on target cells to kill or inhibit pathogenic microorganisms including bacteria, fungi, viruses and the like, improve an immune system of the skin, repair the damaged skin, and achieve an aim of comprehensively treating various skin diseases; and after the use of the NK cell stock solution, a protective film is formed on the surface of the skin, which is beneficial for rapid repair of the damaged skin and prevention of repeated aggravation of an illness.

The present invention relates to retroviral vectors capable of driving the expression of perforin in a cell and a method of expressing recombinant perforin in a cell. The present invention also relates to recombinant perforin polypeptides and nucleic acid molecules derived therefrom and uses thereof. Also encompassed are screening assays employing such recombinant perforin molecules, compounds identified by the screening assays and uses thereof.

The invention discloses an immune cell capable of simultaneously expressing a fusion protein of an intracellular truncated TGF-beta receptor and a cytokine intracellular stimulation domain and a chimeric antigen receptor on a cell membrane. The immune cell can enhance the proliferation capacity of the cell, reduce the influence of a tumor microenvironment, enhance the tumor killing activity of the CAR-T cell and reduce the depletion under the condition of receiving antigen stimulation, so that the negative regulation of the tumor microenvironment on the immune cell is weakened, the depletion of the immune cell is reduced, the tumor killing function of the immune cell is enhanced, and the inhibition of the tumor microenvironment on the immune cell is reduced; by combining with cell factor receptors, corresponding JAK and STAT signal channels are activated, the cells are induced to secrete various active substances such as IFN-gamma, perforin and granzyme, the growth of tumor cells is inhibited, the anti-tumor capability is enhanced, the killing function of immune cells is optimized, and the inhibition of a tumor microenvironment on the immune cells is reduced.

The invention discloses a natural killer cell line (NKL) modified by a human interferon (IFN)-alpha gene, which can stably excrete and express the human IFN-alpha and is established by transfecting the human IFN-alpha gene into an NKL cell. The cell line can be expressed in cells according to the RT-PCR and ELISA detection and identification, and the synthesized IFN-alpha is excreted in extracellular culture supernate. Further research indicates that the NKL cell line modified by the human IFN-alpha gene can effectively kill liver cancer cells, and such anti-tumor effect is related to the expression level improvement of the NKL-IFN-alpha cell, IFN-gamma, Granzyme, TNF-alpha and Perforin genes and proteins. Moreover, the modified NKL cell line is more sensitive to external stimulus. Therefore, the biological characteristics of the cell line are more favorable for the application of clinical adoptive immunotherapy.

The invention provides polypeptide capable of reinforcing killing activity of NK (natural killer) cells and an application of the polypeptide, and belongs to the technical field of life science. The amino acid sequence of the polypeptide capable of reinforcing killing activity of NK (natural killer) cells is as shown in SEQID No.1. After the polypeptide and the NK cells are incubated, the killingactivity of the NK cells to tumor cells can be notably reinforced, and the expression quantity of perforin and CD107a of the NK cells is increased.

The invention discloses an application of a phage Vp670 perforin gene holA. A gene sequence of the holA is as shown in SEQ ID No.1, an encoded amino acid sequence of the holA is as shown in SEQ ID No.2, a gene sequence of cwlQ is as shown in SEQ ID No.3, and an encoded amino acid sequence of the cwlQ is as shown in SEQ ID No.4. According to the application, the holA serving as the perforin gene isfirstly authenticated in the phage Vp670, and the cwlQ is an endolysin gene. According to the application, the HolA is separately expressed in a bacterium and sufficiently kills bacterium cells, a host bacterium does not easily generate tolerance, the HolA can be used for cloning vectors or suicide plasmids, the screening efficiency of positive recombinants can be greatly improved, and laboratoryworkloads are decreased. HolA and CwlQ proteins are collectively expressed in a phage, an expression product can completely lyse the bacterium, a lot of loss of phage contents is forced, and an effective technological mean is provided for mild lysis of microorganisms such as the bacterium and extraction of cell contents.

The invention relates to a preparation method of porcine streptococcus phage perforin in the technical field of biological materials. The preparation method comprises the following steps of: (1) purifying to obtain a porcine streptococcus 2 type virulent phage and extracting the DNA of the phage; (2) amplifying perforin genes by applying a PCR (Polymerase Chain Reaction) through using the DNA obtained in the step (1) as a template; (3) carrying out prokaryocyte expression on the DNA obtained by amplifying in the step (2) to obtain fusion protein; (4) purifying the fusion protein and then carrying out renaturation to obtain the perforin. The purified and activated perforin can be used for cracking multiple porcine streptococcocci in vitro or improving the cracking efficiency of lyase.

The invention provides a method for performing quantitative determination to immunity-relating cytokine in peripheral blood, the method can be applied for specifically detect the cytokine gene expression (PFR) from 2ml of peripheral blood, and no any stimulation is needed before detection. The method can detect the cytokine gene expression under physiologic status, thus can be used for determining early stage fatigue, preventing decrease of body immunity function caused by exercises of excess sport load. The invention also provides a corresponding detection kit.

The invention discloses an application of quercitrin in preparation of a human gamma delta T cell proliferator, and relates to the technical field of medicines. Quercitrin can remarkably promote proliferation of human gamma delta T cells and promote expression of perforin and granzyme B, so that the quercitrin is of certain dose dependency. The gamma delta T cell treated by the quercitrin can promote expression of p-ERK (Extracellular signal-Regulated Kinase)1 / 2 and p-Akt in active form and promote expression of an anti-apoptoasis protein Bcl-2, so that the gamma delta T cell is of certain dose dependency. The invention discovers the novel medical use of quercitrin and provides experimental references to development of tumor immunotherapy medicines.

A novel cell type has been generated that has both Thl characteristics and cytolytic activity. These Thl / killer cells are CD4+ cells purified from peripheral blood and manipulated to have Thl characteristics such as production of IFN-gamma combined with cytolytic activity similar to cytotoxic T-cells (CTL). The CTL activity is targeted toward diseased cells, not normal cells. The cytolytic activity of the Thl / killer cells is mediated by Granzyme B-Perforin mechanism and results in apoptotic death of diseased cells. Methods of producing and using these Thl / killer cells include isolating CD4+ cells from peripheral blood, activating the CD4+ T- cells to form Thl / killer cells and administering these Thl / killer cells with the cytolytic activity to a patient wherein theThl / killer cells are allogeneic to the patient.

The invention relates to the technical field of biological cells, in particular to an immune cell and application thereof. A chimeric antigen receptor is expressed on the cytomembrane of the immune cell, and in addition, an immune checkpoint antibody protein fused with a cytokine receptor is also expressed on the cytomembrane. The expression CAR cell of the immune checkpoint antibody protein fused with the cytokine receptor is also expressed on the cytomembrane; through the immune checkpoint antibody protein which is fused with the cytokine receptor and is expressed on the immune cytomembrane, two negative feedback approaches PD-1 and CTLA-4 are blocked so as to weaken negative regulation and control of a tumor microenvironment for the immune cell, reduce exhaustion of the immune cell, enhance the tumor killing function of the immune cell and reduce inhibition of the immune cell by the tumor microenvironment; and in addition, through binding with the cytokine receptor, corresponding JAK and STAT signal channels are activated, and the cells are induced to secrete various active substances, including IFN-[gamma], perforin, granzyme and the like so as to inhibit growth of tumor cells, enhance an anti-tumor capacity, reduce the exhaustion and reduce the inhibition of the immune cell by the tumor microenvironment.

The present invention relates to new recombinant humanized immunotoxin hS83-P34, which contains fusion protein comprising the antigen associated molecule of humanized cytotoxic T cell antigen 4 and human porforin channel forming active molecule. The immunotoxin of the present invention has strict targeting, less tendency of damaging health cell, less toxic side effect, no need of internalization and intracellular transport, high acting efficiency, less likelihood of generating corresponding antibody, weak immunogenicity and less tendency of generating drug resistance. It may be used in treating some tumors and autoimmune diseases and suppressing immunologic rejection reaction in organ transplantation.

The invention discloses a preparation kit and preparation method of a cell group mainly comprising CD4<+>T cells and a Th1 cell subset, a use of the Th1 cell subset in preparation of an anti-tumor cell preparation, and belongs to the field of biotechnology. The preparation method utilizes a CD4 monoclonal antibody and a culture bottle made of polystyrene, polyvinyl chloride or polyethylene to realize preparation of the cell group mainly comprising CD4<+>T cells, and utilizes two cytokine combination to realize preparation of the Th1 cell subset. Through use of the kit and preparation method provided by the invention, high-ratio CD4<+>T cells in a single karyocyte are obtained. The obtained Th1 cell subset has strong immunocompetence, can secrete a large amount of cell granzyme B, perforin, IFN-gamma and TNF-alpha and has substantial anti-tumor activity.

The invention provides a novel target B7S1 (B7 family member 1) for treating liver cancer. The target reduces activity of cytotoxic T lymphocyte in liver cancer. An inhibitor targeting the B7S1 can promote CD107a expression of the surface of CD8+tumor infiltrating lymphocyte and promote expression of Granzyme B, perforin or IFN-gamma, thereby treating or relieving the liver cancer.

Compounds of formula (1a) and pharmaceutically acceptable salts, solvates, and hydrates thereof and related methods of modulatin perforin activity on a cell: wherein Ring A is selected from a 6-10 membered aryl, 5-6 membered cycloalkyi, 5-6 membered heteroaryl or 5-6 membered heterocyclyl, wherein the heteroaryl and heterocyclyl rings comprise at least one heteroatom selected from N, O or S; and wherein the aryl, cycloalkyi, heteroaryl or heterocyclyl rings are optionally substituted with 1 to 3 substituents selected from halo, nitro, —C1-Cealkyl, —C1-Ceaminoalkyl, —C1-C6hydroxyalkyl, -haloC1-C6alkyl, —C1-C6alkoxyl, -haloC1—C<aIkoxyl, heteroaryl, aryl, hydroxyl, —C(0)Ci-C6alkyl, —OC(0)Ci-C6alkyl, —CH2OC(O)CrC6alkyl, —C(O)OC1, —C6alkyI, —NHC(O)C1, —C6alkyl, —NHS(O)2C1-C6alkyl, —S(O)2C1-C6alkyl, —S(O)2NH2, and —C(O)NJJ; Ring B is a 6-10 membered arylene or a 5-6 membered heteroarylene comprising at least one heteroatom selected from N, 0 or S; and wherein the aryl or heteroaryl is optionally, substituted with one or more substituents selected from —NJJ, —OJ, halo, C1-C6alkyl, -haloC1—C6alkyl, —C1-C6alkoxy, -haloC1-C6alkoxyl, and —C(0)NJJ; Ring C is selected from a 5-10 membered heteroarylene or a 5-10 membered heterocyclene, each comprising at least one heteroatom selected from N, S and O; Ring D is an optionally substituted benzofused 9-11 membered heterocyclyl or optionally substituted ben2ofused 9-11 membered heteroaryl comprising at least one heteroatom selected from N or O; L is a linker selected from branched and unbranched C1-C4 alkylene, —S(0)2-NH—, —C(0)-NH—, —NH—C(0)-NH—, —S(0)2-NH—C(0)-NH—, —S(0)2-NH—C(0)- and —CH═CH—; wherein Rings B and C, and Rings C and D, are connected to each other via a C—C bond at any of the available C atoms on each respective ring; and J in each occurrence is independently selected from H, optionally substituted C1-C6alkyl or optionally substituted haloC1-C6alkyl.

The invention discloses an immune cell for expressing a cytokine receptor fusion type chimeric antigen receptor and application of the immune cell. A cytokine receptor fusion type chimeric antigen receptor is expressed on a cell membrane of the immune cell, and a cytokine receptor intracellular domain is fused in an intracellular domain structure of the cytokine receptor fusion type chimeric antigen receptor. The immune cell can enhance the proliferation capacity of the cell, reduce the influence of a tumor microenvironment and enhance the tumor killing activity of the immune cell under the condition of receiving antigen stimulation, so that the negative regulation of the tumor microenvironment on the immune cell is weakened, the exhaustion of the immune cell is reduced, and corresponding JAK and STAT signal channels are activated by combining with a cytokine receptor, the cells are induced to secrete various active substances such as IFN-gamma, perforin and granzyme, so that the growth of tumor cells is inhibited, the anti-tumor capability is enhanced, and the inhibition of a tumor microenvironment on immune cells is reduced.

The present application provides a composition comprising NK cells and plant-derived extracellular vesicles. The extracellular vesicles can act as NK cell activators to stimulate NK cells to release proteins, such as perforin and / or granzyme, thereby exerting cytotoxicity. The plant-derived extracellular vesicles overcome the problem that the ability of NK cells to attack cancer cells is reduced in the past, enhances the ability of the NK cells to attack the cancer cells, and improves the treatment effect of the NK cells on cancers.

Methods and systems for diagnosing, providing prophylaxis, and treating one or more age-related neurodegeneration or pathological cognitive impairment are provided. An increased presence of CD103+ resident memory CD8+ T cells (CD8+ TRM) can be detected in blood sample obtained from the human subjects with one or more symptoms of loss of short-term or long-term memory, decreased ability to maintain focus, and decreased problem-solving capacity. An increased presence of CD103+ CD8+ TRM can be compared to a value obtained from one or a pool of healthy human subjects with none of the one or more symptoms. One or more of a therapeutically effective amount of: an inhibitor of cluster of differentiation (CD103), an inhibitor of perforin-1, and an inhibitor of interferon gamma (IFNγ) can be administered as treatment of age-related neurodegeneration or pathological cognitive impairment. Pathological neurodegeneration can include Parkinson's disease, multiple sclerosis, or Alzheimer' s disease.

The present invention relates to T cells, in particular a non-activated T cell, comprising an exogenous nucleic acid encoding a T cell Receptor (TCR) specific for a virus. An embodiment of the invention is directed to a non-activated (resting) T cell expressing Hepatitis B virus (HBV) envelope s183-191 TCR capable of inhibiting viral replication and which shows reduced expression of perforins and / or granzymes in response to stimulation as compared to an activated T cell expressing the said TCR. Also encompassed are methods for producing such cells, compositions, pharmaceutical compositions and kits comprising such cells and medical uses thereof.

The invention discloses a method for culturing peripheral blood containing CIK cells, and belongs to the technical field of biology, and the culture method for peripheral blood containing CIK cells comprises the following steps: step 1, extracting peripheral blood and culturing mononuclear cells; and step 2, culturing the CI K cells. The CD28 monoclonal antibody is added in the culture process of the CI K cells, and the CD28 can be used for increasing the activation of the T cells, promoting the generation of the T cells IL-2 and the proliferation of the antigen-specific polyclonal T cells, obviously increasing the proliferation multiple of the CI K cells and improving the reproductive capacity of the CI K cells; IL-1alpha, IL-15 and IL-21 stimulating factors are also added to induce the expression of perforin, granzyme B and CD107a in the cultured CIK cells, so that the cytotoxic activity of the CIK cells is improved.

The invention relates to the field of immunotherapy, in particular to application of IFI35 in preparation of a medicine for preventing or treating colorectal cancer. A colorectal cancer animal model shows that IFI35 can directly regulate CD8 + T cell infiltration and enhance immune response; the method comprises the following steps: analyzing the change of CD8 < + > T cells and T cell functional cytokines (IFN-gamma, Granzyme B and Perforin) in a tumor microenvironment by using a flow cytometry and an immunohistochemical method; the animal model verifies the curative effect of the IFI35 recombinant protein combined with other immunotherapy means. The completion of the invention clarifies that IFI35 is a key molecule affecting the infiltration and function of CD8 + T cells in tumors, and provides a new idea and a potential target for immunotherapy.

The invention relates to a preparation method and application of an engineered exosome for drug targeted delivery. Messenger RNA (mRNA) carrying miRNA or encoded toxic protein (such as perforin or granzyme) capable of inducing specific cell apoptosis is loaded into the exosome. The secreted exosome loaded with specific nucleic acid cargo is collected and purified. A molecular switch encryption device using target cell characteristic miRNA as a triggering signal is preset in 5'-UTR of a target gene, so that the miRNA or mRNA does not have the function in host cells. When the engineered exosome is used as a carrier for conveying a carrier-effector (i.e., the miRNA or toxic protein mRNA capable of inducing target cell apoptosis) to the target cells, the specific miRNAs in cancer cells are used as a triggering switch for starting the release of the effector (miRNA or toxic protein); the apoptosis program is started, so that the target cancer cells are killed.

The invention belongs to the field of biological medicine and the technical field of cell therapy, and particularly relates to a method for efficiently amplifying NK and transfecting NKG2D activated NK cells in vitro. The method comprises the following specific steps: optimizing an NK cell in-vitro efficient amplification technology and detecting NK immune biological characteristics; the method comprises the following steps: detecting mRNA expression of NKG2D ligands MICA and ULBP 1, 2 and 3 in tumor cells and tumor tissues by using an RT-PCR method; performing construction of an NKG2D cloning vector; performing construction of an NKG2D eukaryotic expression vector; expressing and identifying the eukaryotic expression vector in CHO cells; the eukaryotic expression vector transfects NK cells and influences the biological functions of the NK cells. The invention provides a method for constructing a pEGFP-N1 / NKG2D eukaryotic fluorescent expression vector by using a genetic engineering technology, transfecting NK cells with the constructed pEGFP-N1 / NKG2D plasmids, detecting the proliferation situation of the NK cells before and after NKG2D transfection and the killing effect on tumor cell lines by using an MTT method, detecting the mRNA expression level of related killing molecules IL-2, TNF-a, Perforin and TWEAK by using an RT-PCR method. The method for efficiently amplifying the NK in vitro and transfecting the NKG2D activated NK cells, which is a new thought, is provided for tumor treatment.

A novel cell type has been generated that has both Th1 characteristics and cytolytic activity. These Th1 / killer cells are CD4+ cells purified from peripheral blood and manipulated to have Th1 characteristics such as production of IFN-gamma combined with cytolytic activity similar to cytotoxic T-cells (CTL). The CTL activity is targeted toward diseased cells, not normal cells. The cytolytic activity of the Th1 / killer cells is mediated by Granzyme B-Perforin mechanism and results in apoptotic death of diseased cells. Methods of producing and using these Th1 / killer cells include isolating CD4+ cells from peripheral blood, activating the CD4+ T-cells to form Th1 / killer cells and administering these Th1 / killer cells with the cytolytic activity to a patient wherein the Th1 / killer cells are allogeneic to the patient.