39 results about "Perforin" patented technology

The invention discloses a computer device and a computer readable storage medium for evaluating the immune reconstitution state of an HIV infected person after ART. Belongs to the field of computer information. The technical problem to be solved by the invention is how to evaluate the immune reconstruction state of an HIV infected person after ART. The invention provides a computer program stored in a memory in a computer device. The computer program comprises the following steps: S1, receiving sample data, namely the proportion of the number of cells expressing PD-1 in CD8 + T cells of peripheral blood mononuclear cells of an HIV infected subject after ART and the proportion of the number of cells expressing at least one of the following five factors: CD38, HLA-DR, CD57, PERFORIN and CD107A; s2, obtaining a predicted value Y from the sample data through a model formula, and outputting an immune reconstruction condition according to the predicted value Y and a pre-judgment condition; the method can accurately and efficiently determine whether the immune reconstruction state of the HIV infected person subjected to antiviral treatment is good or not.

The invention relates to a method for enhancing target cell removal capacity of immune cells and application. The method comprises the following steps: recombining CD58 and TP53 genes with a granzyme B gene so as to obtain recombinant genes NFAT-GZMB-CD58 and NFAT-GZMB-TP53, and meanwhile, adding a transcription factor NFAT binding site into a regulation region of the obtained recombinant genes. Then, the two recombinant genes are directionally inserted into specific gene loci of immune cells through a gene editing technology. Expression of the two recombinant genes is regulated and controlled by an immune cell activation signal pathway, and translation products of the two recombinant genes can be delivered to target cells of immune cells through a granzyme-perforin pathway, so that the scavenging capacity of the immune cells to the target cells is enhanced, and apoptosis and escape of the target cells are inhibited. The method can be applied to the field of cell therapy.

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.

The invention belongs to the technical field of biology, and particularly relates to perforin derived from staphylococcus warneri mild bacteriophage and application. Perforin has a nucleotide sequence as shown in SEQ ID NO: 1. The perforin is a key gene for bacteriophage splitting growth and plays an important role in the splitting process of host bacteria. Therefore, the research aims to verify whether the perforin of which the coding site is positioned on a staphylococcus warneri mild bacteriophage genome can be used as a cell lysis gene and can be used as a potential antibacterial substance or not, and a theoretical basis is provided for prevention and treatment of gram-negative pathogenic bacteria.

The invention discloses a method for enhancing the immune function of bone marrow precursor cells by using a 33-site monoubiquitin molecule, and relates to the field of genetic engineering technology. Bone marrow precursor cells BMPCs (Bone Marrow Precursor Cells) are isolated from mouse bone marrow, and the bone marrow precursor cells are pretreated with a 33-site monoubiquitin molecule, and then adoptively immunized in vitro, and finally subjected to enzyme-linked immunosorbent assay and flow cytometry experiments. The experiment shows that the immune function of the treated bone marrow precursor cells is significantly enhanced, and their antigen-specific CTL induction ability is also enhanced, and their ability to kill tumor cells is enhanced; the 33-site monoubiquitin molecule can increase the ability of bone marrow precursor cells to transfer intracellular lymphocytes to produce perforin and granzyme B. Bone marrow precursor cells enhanced by the 33-site monoubiquitin molecule can be used in the preparation of tumor immunotherapy drugs.

The present invention relates to a self-cleaving engineered bacterium comprising a metabolic pathway for producing polyhydroxyalkanoate (PHA); the perforin gene and the endolysin gene are expressed under the control of a promoter. According to the method, on the basis of not influencing the dry cell weight and PHA yield of the engineering strain, the self-splitting of the engineering strain after PHA accumulation is completed is realized, the downstream extraction step of PHA biosynthesis is greatly simplified, and the extraction time is shortened.

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.

This invention belongs to the field of biotechnology, specifically relating to a perforin derived from *Staphylococcus warwickiana* temperate bacteriophage and its applications. The perforin has the nucleotide sequence shown in SEQ ID NO:1. This perforin is a key gene for phage lysis and growth, playing a crucial role in the lysis of host bacteria. Therefore, this study aims to verify whether the perforin, whose coding site is located on the genome of *Staphylococcus warwickiana* temperate bacteriophage, as a cell lysis gene, can serve as a potential antibacterial substance, providing a theoretical basis for the prevention and control of Gram-negative pathogens.

The application provides an immune cell culture kit, a culture method and application of gamma delta T cells, and the kit comprises a basic culture medium, a recombinant antibody, a cytokine and a cell proliferation promoting additive, the basic culture medium comprises an RPMI-1640 culture medium, and the cell proliferation promoting additive comprises 1-thioglycerol, taurine, beta-cyclodextrin, thymidine, alpha-lipoic acid, beta-mercaptoethanol and bisphosphonate. The gamma delta T cells obtained by the culture kit have high purity and activity, and have good tumor killing activity. The gamma delta T cells obtained by the method have a high positive rate of 100% of perforin expression.

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 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.

The present invention provides compositions and methods for isolating, manipulating mammalian MHC class I restrictive antigen-specific regulatory T cells, and for therapeutic and other purposes. The regulatory T cells may be characterized as CD8 + cells that specifically inhibit the response of autoreactive and / or pathogenic CD4 + T cells by cytotoxic mechanisms, including but not limited to Perforin, other components of the Perforin / granzyme apoptosis pathway, and the like. The regulatory T cells have antigen specificity but are not activated by the same antigen as the autoreactive and / or pathogenic CD4 + T cells. In a human body, the regulatory T cells express an inhibitory KIR protein, such as one or more of KIR2DL2, KIR2DL3, and KIR3DL1.

The invention belongs to the technical field of pharmaceutical compositions for removing senescent cells, and discloses a pharmaceutical composition for removing senescent cells and a preparation method and application thereof, and the pharmaceutical composition for removing senescent cells contains NK cells and fisetin. According to the pharmaceutical composition for removing senescent cells, the fisetin and the NK cells are combined for use, on one hand, the fisetin can enable the NK cells to easily recognize senescent cells; on the other hand, fisetin can relieve inhibition of SASP factors on NK cell activity, enough amount of cytotoxic particles such as perforin and granzyme are synthesized and released in NK cells, the killing efficiency on senescent cells is high, fisetin can directly trigger an apoptosis pathway of the senescent cells, and the effect of killing the senescent cells is achieved. The NK cells release cytotoxic particles to further accelerate death of the senescent cells, and fisetin and the NK cells have a synergistic effect, so that the senescent cell clearing efficiency is high.

Compositions and methods are provided for isolating, manipulating and using for therapeutic and other purposes, mammalian, MHC Class I restricted, antigen-specific regulatory T cells. The regulatory T cells can be characterized as CD8+ cells that specifically suppress the responses of self-reactive and / or pathogenic CD4+ T cells by cytotoxic mechanisms including, without limitation, perforin, other components of the perforin / granzyme apoptosis pathway, etc. The regulatory T cells are antigen-specific, but are not activated by the same antigen as the self-reactive and / or pathogenic CD4+ T cells. In humans the regulatory T cells express inhibitory KIR proteins, e.g. one or more of KIR2DL2, KIR2DL3, and KIR3DL1.

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.

The present invention belongs to the field of life science and technology. More specifically, it relates to a polypeptide for enhancing the killing activity of NK cells and its application. The polypeptides for enhancing the killing activity of NK cells in the present invention are LL-37 polypeptide and Nigroain-E1 polypeptide. After co-culturing the above polypeptides with NK cells, Nigroain-E1 polypeptide can inhibit the cytotoxicity of LL-37 polypeptide and does not affect the biological activity of LL-37 polypeptide. In the presence of Nigroain-E1 polypeptide, LL-37 polypeptide can enhance the killing activity of NK cells against tumor cells. After treatment with the combination of the above two polypeptides, it can effectively promote the expression level of perforin in NK cells, thereby enhancing its killing activity against tumor cells.

The application relates to application of arachidonic acid inhibitors in preparation of drugs for improving tumor immune microenvironment. + The arachidonic acid inhibitors improve the ratio and expression amount of perforin and IFN-gamma secreted by T cells and improve the ratio and expression amount of IFN-gamma and IL-17A secreted by Treg cells.

The invention discloses application of a gene modified NK cell in tumor treatment, and belongs to the technical field of biology. The recombinant NK cell for expressing the human perforin-collagenase fusion protein is constructed through a CRISPR-Cas9 technology. The killing activity of a common NK cell to a human breast cancer cell MDA-MB-231 is 59.47%, the killing activity of a recombinant NK cell is 85.71%, and the killing activity is improved by 44.12%. A treatment test on a breast cancer model mouse shows that the average volume of a common NK cell group mouse breast tumor is 117.02 mm, and the NK cell infiltration rate is 15.28%; the mouse breast tumor of the recombinant NK cell group is obviously reduced, the average volume is 86.03 mm, and the NK cell infiltration rate is increased to 22.58%. The infiltration rate and killing activity of the recombinant NK cell on solid tumors are remarkably improved, and the recombinant NK cell can play an effective role in tumor treatment.

The present disclosure relates generally to pharmaceutical formulations (e.g., lyophilized formulations or topical formulations) comprising a perforin inhibitor (e.g., a benzenesulfonamide). In particular embodiments, the pharmaceutical formulations of the present disclosure are useful for treating diseases or disorders associated with undesirable perforin activity.

The invention belongs to the technical field of bacteriophages, and discloses a recombinant bacteriophage for displaying bacteriophage perforin and a preparation method and application thereof.The preparation method of the recombinant bacteriophage comprises the following steps that S1, a bacteriophage genome is used as a template, a fusion primer is used for amplifying a bacteriophage perforin coding gene sequence, and a recombinant bacteriophage is obtained; obtaining a bacteriophage perforin coding gene sequence, and carrying out enzyme digestion and recovery on an amplification product and an expression vector; s2, connecting a bacteriophage perforin coding gene sequence with a homologous arm with the expression vector skeleton subjected to enzyme digestion, and transforming into Escherichia coli to obtain recombinant Escherichia coli; and S3, carrying out phage rescue on the recombinant escherichia coli to obtain the recombinant phage displaying the phage perforin. The lysis spectrum of the recombinant bacteriophage prepared by the method is greatly widened, various gram-positive bacteria and mycoplasmas can be subjected to cross-species lysis, the lysis efficiency is also improved, and the recombinant bacteriophage has important significance and value in the aspects of antibacterial treatment, clinical application and the like.

The invention discloses application of ganoderenic acid A in enhancing the killing function of T lymphocytes. The ganoderenic acid A is lanolin alkyne alkyl type triterpenoid, and it is proved that the ganoderenic acid A can promote T lymphocytes to release granzyme, perforin and interferon within the concentration range of 5-10 [mu] M, and then the killing function of the T lymphocytes is promoted. The early foundation is laid for the ganoderenic acid A to become an immunoregulation small molecule medicine in the future, and it is indicated that the ganoderenic acid A is expected to become a recognized XinJin immune activator.

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 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.

The invention relates to the field of metabolic engineering, in particular to an engineered microorganism as well as a construction method and application thereof. A first gene and a second gene are simultaneously expressed in the engineered microorganism, the first gene is a perforin protein coding gene with a nucleotide sequence as shown in SEQ ID NO: 1, and the second gene is a coding gene of hydrolase protein with a nucleotide sequence as shown in SEQ ID NO: 2. The engineered microorganism can complete the extraction of intracellular target products, reduce or completely replace the addition of exogenous enzyme preparations, and significantly reduce the use cost of enzyme.

The invention discloses a method for enhancing the immune function of dendritic cells by using 11-site single ubiquitin molecules, and relates to the technical field of gene engineering. The invention discloses a 11-site single ubiquitin molecule which is a ubiquitin molecule only retaining 11-site lysine, and relates to the technical field of gene engineering. Dendritic cells are separated from mouse bone marrow, the bone marrow-derived dendritic cells are pretreated with 11-site single ubiquitin molecules, then in-vitro adoptive immunization is performed, and finally an enzyme-linked immunosorbent assay spot experiment and a flow cytometry experiment are performed. Experiments prove that the antigen-specific CTL (cytotoxic T lymphocyte) inducing capacity of the treated dendritic cells is enhanced; the 11-site single ubiquitin molecule can enhance the phagocytic ability of the dendritic cells to the antigen and the ability of the dendritic cells to transvey lymphocytes in the receptor to generate perforin and granzyme B; the mechanism of the 11-site single ubiquitin molecule for enhancing the presentation and immune function of the dendritic cells on the antigen is mainly realized by a proteasome degradation pathway.

The invention relates to construction of an engineering strain of high-yield soluble bacteriophage perforin Hol41 protein, a high-yield expression method, application and the engineering strain, fungi are used as host cells, and expression plasmids capable of secreting signal peptides are adopted to carry out free expression on bacteriophage perforin Hol41 genes, so that the engineering strain of high-yield soluble bacteriophage perforin Hol41 protein is obtained. Through the construction, the induced expression quantity of the soluble bacteriophage perforin Hol41 protein is greatly improved.

The invention discloses a nano CAR T cell as well as a preparation method and application thereof. The nano CAR T cell comprises a CAR T cell, and the surface of the CAR T cell is connected with lipid nanoparticles for activating endogenous immunity. The nano CAR T cell prepared by the invention can be rapidly homed to a tumor part by intravenous injection, the CAR T cell recognizes tumor antigen to be activated and secretes granzyme and perforin, and the lipid nanoparticles respond to CAR signal release, target in-tumor DC cells, promote maturation of the DC cells and uptake tumor-associated antigens released by CAR T cracking tumor, so that the CAR T cell can be rapidly homed to the tumor part. And a plurality of antigen-specific T cells are generated in vivo to cooperatively kill antigen-negative tumor cells.

The invention discloses a method for reducing the immune function of dendritic cells by using 27-site single ubiquitin molecules, and relates to the technical field of gene engineering. The 27-site single ubiquitin molecule is a ubiquitin molecule only retaining 27-site lysine, dendritic cells are separated from mouse bone marrow, the bone marrow-derived dendritic cells are pretreated with the 27-site single ubiquitin molecule, then in-vitro adoptive immunization is carried out, and finally an enzyme-linked immunosorbent assay spot experiment and a flow cytometry experiment are carried out. Experiments prove that the antigen presenting capability of the treated dendritic cells is obviously reduced, and the antigen-specific CTL inducing capability of the treated dendritic cells is also reduced; the 27-site single ubiquitin molecule can reduce the capability of generating IFN-gamma, perforin and granzyme B by lymphocytes in a dendritic cell transfer receptor. The 27-site single ubiquitin molecule inhibits splenocytes of a transfer receptor to generate perforin and granzyme B through a lysosome way.

The invention discloses application of gamma-aminobutyric acid in preparation of a synergist of an immune checkpoint inhibitor, and belongs to the technical field of tumor treatment. Compared with independent treatment, combined administration of the GABA and the immune checkpoint inhibitor can significantly inhibit tumor growth, increase the infiltration proportion of CD8 + T cells in tumor tissues, and increase the proportions of IFN-gamma +, TNF-alpha + and Perforin + CD8 + T cells, so that the cytotoxic function of the T cells is enhanced. The results show that the GABA can promote the anti-tumor immune response of an organism by adjusting immune cell composition and T cell functions in a tumor microenvironment, so that the treatment effect of the immune checkpoint inhibitor is improved, and a new theoretical basis and application prospect are provided for the GABA as an auxiliary strategy for improving the tumor immunotherapy curative effect.

This invention provides a BiTE for targeted immunotherapy of tumors, its preparation method, and its application, relating to the field of biopharmaceutical technology. The BiTE is obtained by conjugating a nanobody to a fusion protein. The nanobody contains the amino acid sequence SEQ ID No. 3 at its C-terminus and has the function of targeting the human epidermal growth factor receptor on tumor cells. The fusion protein also targets the CD3 receptor on T cells and activates T cells, connecting T cells and tumor cells in time and space, activating T cells to secrete perforin and granzyme B to kill tumor cells, thus exerting a targeted immunotherapy effect. This invention utilizes isopeptide bonds to conjugate the nanobody and the fusion protein, completing the conjugation within 15 minutes with a conjugation efficiency of 100%, and extending the in vivo half-life of the nanobody by 6 times.