60 results about "Cellular immunity" patented technology

The application provides a cell fixing solution for a myasthenia gravis antibody spectrum (IgG) detection kit (cellular immunofluorescence method) and a product and application thereof, and relates to the technical field of in-vitro detection. The cell fixing solution provided by the application is composed of polyoxymethylene, formaldehyde, acetic acid, raffinose and a buffer solution, can effectively fix the cell morphology, maximally retains the natural spatial conformation and antigen epitope integrity of myasthenia gravis related antigens AChR, Titin, RyR1, MuSK and LRP4, reduces the problems of antigen denaturation, epitope shielding and protein loss in the fixing process, can efficiently and accurately realize the synchronous joint detection of various specific antibodies, and the myasthenia gravis antibody spectrum (IgG) detection kit (cellular immunofluorescence method) prepared by the application has stable detection results, and has important clinical application value and industrial popularization prospect.

This invention relates to the field of biopharmaceutical technology, disclosing a QS-21 saponin adjuvant, its preparation method, and its applications. The adjuvant is a composite liposome comprising a liposome backbone composed of distearate phosphatidylcholine and cholesterol, with QS-21 saponin, monophosphatidyllipid A, and a local anesthetic co-encapsulated therein. This invention solves the common problem of high reactivity and immunogenicity in potent adjuvants by co-encapsulating immunostimulatory and analgesic components on the same nanocarrier, achieving synergistic delivery at the injection site, thereby precisely inhibiting adjuvant-induced pain and swelling. This co-encapsulation structure avoids the potential inhibitory effect of free anesthetics on the immune system, fully preserving the adjuvant's potent humoral and cellular immune-enhancing activity. This invention provides a new technical solution for developing vaccines with both high safety and strong immunogenicity.

Described herein are covalently modified polypeptide antigens with improved immunogenicity and / or stability, as well as compositions, cells, and methods related to the polypeptide antigens. The polypeptide antigens are covalently conjugated to one or more steroid acid moieties to improve their stability and / or to elicit improved cellular immunity or improved cellular and humoral immunity against the antigens after administration to a subject. The steroid acids include bile acids and bile acid analogs that enhance endosomal trapping of cargo by enhancing the enzymatic cleavage of endosomal membrane- ensheathed sphingomyelin to ceramide and / or endosomal escape. The steroid acid moieties can be preconjugated to a peptide, and the steroid acid-peptide moiety is subsequently conjugated to the polypeptide antigen. The peptide can comprise one or more domains that confer additional functionality to the modified polypeptide antigen.

This invention discloses an antigen and vaccine preparation based on dominant Brucella epitopes, and their application, belonging to the field of recombinant subunit vaccine technology. The antigen is composed of CTL epitopes, HTL epitopes, and B-cell epitopes tandemly, and its amino acid sequence is shown in SEQ ID NO.1. Based on screened dominant CTL epitopes, HTL epitopes, and B-cell epitopes of the Omp25 and Omp31 outer membrane proteins, this invention constructs a novel antigen fusion polypeptide. The vaccine prepared based on this polypeptide effectively activates a significant dual immune response, enhancing cellular immunity while inducing the body to produce high levels of specific antibodies. It also provides good protection against organ damage caused by bacterial infection, offering better immune response and protective efficacy against Brucella, laying the foundation for the development of Brucella recombinant subunit vaccines.

The invention discloses a blocking agent for cell immunofluorescence detection by a slide method and a use method of the blocking agent. The blocking agent is a homologous no-load cell lysis solution without transfection plasmids, and the protein concentration is greater than or equal to 30mg / mL. The blocking agent is completely the same as cell cytoplasm and cell membrane components of cells to be detected except for plasmids-expressed antigens, when in use, the blocking agent is pre-mixed with a blood cleaning sample in advance, interfering antibodies except target antibodies in serum can be purified, and meanwhile, due to the fact that no antigen protein expressed after plasmid transfection exists, the blocking agent can be used for detecting the cells to be detected. And the protein concentration is equivalent to the globulin concentration in serum, so that the target antibody cannot be lost in the serum, and the slide method CBA detection sensitivity can be greatly improved.

The invention discloses a CAR-T cell for targeted knockout of a CMPK2 gene, a preparation method of the CAR-T cell and application of the CAR-T cell in solid tumor resistance, and belongs to the technical field of tumor immunotherapy. Experiments prove that the targeted knockout of the CMPK2 gene can improve the capability of killing HepG2 tumor cells in vitro by CAR-T cells; in addition, after targeted knockout of the CMPK2 gene, the killing ability of CAR-T cells on NSG mouse liver cancer solid tumors can be improved, and it is proved that the CMPK2 target has great clinical cellular immunotherapy potential. According to the invention, the problem that the efficacy of the traditional CAR-T cell therapy is reduced in solid tumor treatment is solved, the anti-tumor activity and the treatment effect of the CAR-T cells are remarkably improved, meanwhile, higher safety and applicability are ensured, and the CAR-T cells have important clinical application value and wide development prospects.

The invention belongs to the technical field of gene engineering, and particularly relates to recombinant lactobacillus for expressing clostridium perfringens multiple antigens of targeted dendritic cells as well as a construction method and application of the recombinant lactobacillus. The recombinant lactobacillus contains an expression vector for expressing a fusion gene; the fusion gene comprises sequences for coding and expressing a PgsA '-fimA fusion protein, a LysM-Fba fusion protein, an LP1261-NetB fusion protein, a phi54 targeting element and a Pas escape element; a sequence for coding the PgsA '-fimA fusion protein is shown as SEQ ID NO: 1, a sequence for coding the LysM-Fba fusion protein is shown as SEQ ID NO: 2, and a sequence for coding the LP1261-NetB fusion protein is shown as SEQ ID NO: 3. When the recombinant lactobacillus is used for immunizing animals, mucosal immunity, cellular immunity and humoral immunity reactions can be induced, the immunocompetence of organisms can be comprehensively enhanced, and the recombinant lactobacillus can be used for preparing oral vaccines for prevention and control of chicken necrotic enteritis.

A method for treating a tumor, characterized by administering to an individual having a tumor immune effector cells expressing a receptor recognizing a tumor antigen and gemcitabine. A kit for treating a tumor, characterized by comprising: 1) immune effector cells expressing a receptor recognizing a tumor antigen; 2) gemcitabine; 3) a container for containing the above 1) and 2); and 4) a written notice for treating a tumor using the kit.

The present disclosure relates to methods for determining the magnitude of a subject's immune response against a HIVACAT T-cell immunogen (HTI or “HTI immunogen”) and whether the subject can avoid antiretroviral therapy (ART). These methods are helpful for treating human immunodeficiency virus (HIV) and / or deciding whether to administer, continue or stop antiretroviral therapy in a subject. The present disclosure also relates to antigens, compositions, and kits related to such methods.

A method for treating a tumor, characterized by administering to an individual having a tumor immune effector cells expressing a receptor recognizing a tumor antigen and gemcitabine. A kit for treating a tumor, characterized by comprising: 1) immune effector cells expressing a receptor recognizing a tumor antigen; 2) gemcitabine; 3) a container for containing the above 1) and 2); and 4) a written notice for treating a tumor using the kit.

The present application relates to the technical fields of biological medicine and cellular immunotherapy, and particularly relates to a signal conversion receptor targeting IL-10, an engineered macrophage and application thereof. The signal conversion receptor is composed of an extracellular domain and a transmembrane domain and an intracellular domain derived from TLR9, and the extracellular domain sequentially comprises a signal peptide, a HA tag and a specific binding domain of an IL-10 receptor alpha subunit from N-terminal to C-terminal. The present application further prepares an engineered macrophage SR CAR-M capable of specifically recognizing IL-10 and converting it into a TLR9 activation signal, which can induce macrophages to polarize to M1 type and has excellent phagocytosis and killing capacity for bladder cancer, breast cancer, lung cancer and melanoma cells, and can be used for preparing related tumor treatment drugs, overcoming the common problems of existing cell therapy, such as easy exhaustion, difficult infiltration and easy inhibition in solid tumors, and having significant clinical transformation potential.

The application discloses a mycobacterium tuberculosis tandem DNA vaccine W545 and a preparation method and application thereof. The application constructs a novel mycobacterium tuberculosis DNA vaccine W545 with a multi-antigen immune dominant epitope by connecting epitope genes of Ag85A protein and Ag85B protein antigen and Rv1419, Rv3407 and Rv2628c together in series through genetic engineering technology and connecting the epitope genes to a eukaryotic expression vector pVAX1. The prepared mycobacterium tuberculosis tandem DNA vaccine W545 can significantly enhance specific cellular immune function of mice, mainly stimulates Th1 type immune response, and can make the lesion range of organs and tissues of a mouse tuberculosis model significantly reduced and the lesion alleviated in treatment, and can be used in preparation of drugs or vaccines for preventing or treating tuberculosis.

This invention belongs to the field of biodetection technology, specifically relating to a material for detecting anti-Caspr1 antibodies and its preparation method. The invention constructs an expression plasmid capable of highly expressing the Caspr1 antigen, transfects it into HEK293T cells, and modifies the original antigen sequence during construction to achieve normal membrane expression. Subsequently, a modified cell culture protocol is used to obtain a material for detecting anti-Caspr1 antibodies based on cellular immunofluorescence assay. This invention, based on cellular immunofluorescence, avoids the limitations of flow cytometry, which is complex and requires expensive equipment. By modifying the protein expression mode to normal membrane expression, it avoids cell permeabilization, avoids false negatives caused by antigen conformational changes in enzyme-linked immunosorbent assay (ELISA) and Western blotting, and avoids false negatives caused by transmembrane expression due to transfection with the original Caspr1 sequence.

The present application belongs to the technical field of cellular immunization engineering, and particularly relates to a polypeptide specifically recognizing PSCA and application thereof. The polypeptide provided by the present application specifically recognizes PSCA and has good affinity performance. The polypeptide specifically targeting PSCA provided by the present application can mark PSCA positive cells, combine with PSCA antigen and PSCA positive cells, and can be used as a detection reagent for PSCA molecules and an in-vivo tracing labeling reagent. The polypeptide specifically targeting PSCA provided by the present application can also be used in an extracellular antigen recognition region of a chimeric antigen receptor, and can be used in an immune cell treatment drug targeting PSCA to kill malignant tumors expressing PSCA. The polypeptide specifically recognizing PSCA of the present application is derived from a natural full human antibody, and compared with a mouse-derived antibody, a chimeric antibody and a humanized antibody, the immunogenicity is greatly reduced, and the safety can be maximally ensured in clinical application.

The invention is applicable to the field of gene engineering, and provides a mycobacterium tuberculosis ESAT-6-LS protein nanoparticle, a preparation method and application thereof, the mycobacterium tuberculosis ESAT-6-LS protein nanoparticle is formed by sequence fusion of mycobacterium tuberculosis ESAT-6 protein and LS protein, and the amino acid sequence of the mycobacterium tuberculosis ESAT-6-LS protein nanoparticle is shown as SEQ ID NO: 2 in a sequence table. The ESAT-6-LS protein nanoparticle provided by the invention is non-toxic and free of pathogenicity, can ensure that a vaccine has good biological safety, and is suitable for clinical application. Animal experiments show that after a mouse is immunized by the ESAT-6-LS protein nanoparticle, strong humoral immunity (antibody generation) and cellular immunity (T cell response) can be stimulated at the same time, and it is proved that the recombinant ESAT-6-LS protein nanoparticle vaccine can effectively induce specific immune protection.

This invention belongs to the field of biodetection technology, specifically relating to a material for detecting anti-NF155 antibodies and its preparation method. This invention constructs an expression plasmid capable of highly expressing the NF155 antigen, transfects it into HEK293T cells, and modifies the pCDH vector with enhancers during construction. Subsequently, a modified cell culture protocol is used to obtain a material for detecting anti-NF155 antibodies based on a cell immunofluorescence methodology. This invention uses a cell immunofluorescence methodology to maximize the preservation of the antigen's spatial conformation through cell expression and antigen immobilization, preventing false negatives due to changes in binding sites. This invention selects the lentiviral vector pCDH to avoid the problem of non-specific binding that is difficult to distinguish, as the proteins expressed by common target gene vectors such as pcDNA3.1 may themselves contain antibody binding sites. This improves detection specificity and solves the problem of false negatives.

This invention discloses a porcine CD40L trimer protein, a recombinant Bacillus subtilis strain expressing this protein, and its applications, belonging to the fields of microbial genetic engineering and livestock and poultry biological products technology. This invention utilizes Bacillus subtilis WB800N as a host and pHT43 as a vector to successfully construct a recombinant strain that efficiently secretes and expresses porcine CD40L trimer protein. Soluble expression of the porcine CD40L trimer protein was achieved. The amino acid sequence of the porcine CD40L trimer protein expressed by the recombinant strain is shown in SEQ ID NO:1. Immunological activity verification using this porcine CD40L trimer protein revealed that the CD40L trimer protein expressed in this invention can simultaneously activate humoral and cellular immunity, possessing practical application value as a novel vaccine adjuvant.

The invention relates to the technical field of cellular immunotherapy, and discloses an application of a CD74 positive regulatory T cell in treatment of graft versus host disease, the CD74 positive regulatory T cell is composed of the following components in proportion: in a sorted and purified cell population, the proportion of regulatory T cells with CD4 + CD25 + CD127-phenotype is 85-95%, the proportion of regulatory T cells with CD25 + CD127-phenotype is 1-5%, and the proportion of regulatory T cells with CD24 + CD25 + CD127-phenotype is 1-5%. Wherein the CD74 high-expression subgroup accounts for 60-75% of the total amount of the regulatory T cell, the cell subgroup functional immune molecule combination comprises CTLA4, FOXP3, TIGIT and TNFRSF18, when the CD74 positive regulatory T cell is used for treating graft versus host disease, the CD74 positive regulatory T cell is firstly used for preventive infusion, single infusion is performed on the day of transplantation, the dosage is 5 * 10 < 5 > cells / receptor, and then the CD74 positive regulatory T cell is used for treating the graft versus host disease. The CD74 positive regulatory T cells are used for repeated therapeutic infusion when early aGVHD symptoms occur, graded treatment is carried out, pathological immune response is inhibited to the maximum extent, immune tolerance is promoted, the treatment is carried out once a week and 2-3 times in total, and the dosage of each time is 1 * 10 < 6 > cells / receptor.

The invention belongs to the technical field of genetic engineering, and particularly relates to recombinant lactobacillus plantarum for preventing and controlling chicken necrotic enteritis as well as a preparation method and application of the recombinant lactobacillus plantarum. The invention provides an oral vaccine vector capable of co-expressing various key virulence factor antigens (FimB, NetB, CnaA and FBA) of clostridium perfringens, and by fusing dendritic cell (DC) targeting molecules (nano antibody Phi74), the efficiency of recognizing and taking in the vaccine vector by full-time antigen presenting cells is remarkably improved, so that the immunogenicity is enhanced. When the recombinant lactobacillus is used for immunizing animals, mucosal immunity, cellular immunity and humoral immunity reactions can be induced, the immunity of organisms is comprehensively enhanced, and a safe and efficient oral immunization scheme suitable for large-scale breeding is established for treating chicken necrotic enteritis.

The invention belongs to the technical field of cellular immunotherapy, and particularly relates to a targeted FAP chimeric antigen receptor macrophage as well as a preparation method and application thereof. According to the invention, FAP is taken as a target spot to design a CAR carrier of a specific targeted activated hepatic stellate cell, and a CAR-M cell for treating hepatic fibrosis is constructed. The macrophage expressing the chimeric antigen receptor has a remarkable killing effect on FAP positive cells, and a new thought is provided for clinical treatment of hepatic fibrosis, liver cirrhosis and liver cancer.

The invention provides a combined vaccine based on Th1 / Th2 immune dynamic equilibrium as well as a preparation method and application of the combined vaccine. Subcutaneous delivery of interleukin 12 (IL12) and antigen (such as ovalbumin OVA) mRNA is adopted in the early stage of tumor occurrence, so that specific antigen recognition and T cell immune response of an organism are activated; when immune response enters a function decline stage, the tumor microenvironment is regulated through intratumor delivery of interleukin 4 (IL4) mRNA, the metabolic activity of T cells is maintained, and the depletion of the T cells is delayed. Through space (subcutaneous / intra-tumor) and time (early stage / middle stage) separated delivery of different immune factors, 'start-steady state maintenance 'type immune stylization regulation and control are realized, antigen-specific CTL reaction and persistent anti-tumor effect are remarkably enhanced, and the problem of immune bias caused by independent use of the antigen-specific CTL reaction and the persistent anti-tumor effect is solved.

The invention provides a vaccine adjuvant, a protein used by the vaccine adjuvant and a preparation method of the protein. The technical problem to be solved is how to provide an effective and nontoxic vaccine adjuvant. The invention discloses a protein. The protein is any one of the following items: A1) a protein with an amino acid sequence as shown in a sequence 2; a2) a protein which is obtained by substitution and / or deletion and / or addition of amino acid residues of the protein in A1), has 80% or more of identity with the protein in A1) and has the same function as the protein in A1); and A3) a fusion protein obtained by connecting the N terminal or / and C terminal of A1) or A2) with a protein tag. The invention discloses an influenza virus adjuvant which is named as protein Ad-20, and the protein Ad-20 serving as the influenza virus adjuvant can stimulate humoral immunity and cellular immunity, has the functions of stimulating maturation of B cells and promoting the B cells to secrete antibodies in advance, is used for enhancing immunogenicity and immune memory of virus vaccines, is non-toxic and has the effect higher than that of an aluminum adjuvant; and a new auxiliary means is provided for research and development of effective vaccines.

The invention relates to the technical field of cellular immunotherapy, in particular to a preparation method of universal CAR-T (Chimeric Antigen Receptor-T) cells for targeting CD19 mediated by umbilical cord blood, and provides a standardized and large-scale preparation scheme aiming at the problems of long preparation period, high cost, large immune risk, limited applicable population and the like of the existing autologous CAR-T cells. The method comprises the steps of umbilical cord blood collection and storage, T cell separation and purification, CD19-targeted CAR gene construction and lentivirus transduction, TCR and HLA-I type molecule knockout through gene editing, CAR-T cell amplification and quality control and the like. The advantages of low immunogenicity and high multiplication capacity of the umbilical cord blood T cells are utilized, an optimized gene modification and culture system is combined, batch production of the CAR-T cells is achieved, the preparation period is shortened, the cost is reduced, adverse reactions such as GVHD and CRS are reduced, the applicable crowd for treatment is expanded, and an efficient and safe new strategy is provided for treatment of malignant B cell diseases.

The invention relates to an MPLA-QS-21 loaded polymer solid lipid composite nano adjuvant as well as a preparation method and application thereof in prevention of virus infection. The MPLA-QS-21-loaded polymer lipid composite nano adjuvant is solid lipid spherical nano particles which entrap MPLA and QS-21 and are formed by taking a polylactic acid degradable polymer, cholesterol and cationic lipid as materials in an O / W mode, and an antigen is carried on the surface of the nano particles in an electrostatic adsorption mode to form a nano adjuvant vaccine. The MPLA-QS-21 loaded polymer solid lipid composite nano adjuvant vaccine can promote the activation of antigen presenting cells, quickly activate cellular immunity and humoral immunity and secrete high-level antibodies and cytokines, so that the purpose of effectively preventing virus infectious diseases is achieved, and the MPLA-QS-21 loaded polymer solid lipid composite nano adjuvant vaccine is an effective delivery system for enhancing antigen immunogenicity.

The invention provides a method for preparing gamma delta T cells derived from induced pluripotent stem cells and application of the gamma delta T cells, and belongs to the technical field of cellular immunotherapy. According to the method, the induced pluripotent stem cells can be efficiently differentiated and amplified in a 3D suspension culture mode, and a scheme for preparing universal spot type gamma delta T cells in a large scale can be achieved; the method provided by the invention has the advantages of short differentiation period, high purity, no need of trophoblasts for differentiation, stability of the differentiation process, and wide industrialization and clinical application prospects, and can be used as a process platform for large-scale production of gamma delta T cells with homogenized quality.

The present application relates to a kind of nucleotide, vaccine comprising it and composite vaccine adjuvant.The sequence of the nucleotide is shown as SEQ ID No:1.Relative to conventional technology, the beneficial effects of the present application include: the present application adopts new optimized sequence, provides a kind of nucleotide, the nucleotide can be as CpG oligonucleotide adjuvant, induces vaccine antigen to produce stronger humoral immunity, cellular immunity level and anti-tumor immune effect etc., has obvious clinical application value and vaccine development value.Moreover, using the immunoadjuvant of the present application helps to reduce the dose of clinical use vaccine, improves safety.

The present application relates to the technical field of cellular immunotherapy, and particularly relates to a kind of targeting PSMA chimeric antigen receptor immune cells and preparation method and application thereof.A kind of targeting PSMA chimeric antigen receptor modified immune effector cell, the chimeric antigen receptor includes antigen binding domain, hinge region, transmembrane domain and signal transduction domain, and the antigen binding domain is anti-PSMA single-chain antibody.The chimeric antigen receptor modified immune effector cell provided in the present application can specifically recognize and combine the antigen binding domain of PSMA, and the prepared chimeric antigen receptor macrophage has specific killing effect on the target cells positive for PSMA protein, and can be used for preparing tumor drugs for treating or preventing PSMA positive.

The invention discloses a genetically engineered NK cell and application thereof in treatment of metabolic dysfunction related fatty liver diseases, and relates to the field of cellular immunotherapy and genetic engineering. The invention provides a genetically engineered NK cell. The NK cell is genetically engineered to overexpress or heterologous express a CD6 molecule. The invention provides application of the genetically engineered NK cell in preparation of drugs for preventing or treating metabolic dysfunction related fatty liver diseases and progressive diseases thereof. The modified NK cell disclosed by the invention can highly express CD6, can specifically recognize and target pathological hepatocytes which highly express ALCAM in MASH / hepatic fibrosis through a CD6-ALCAM axis, and is clear in treatment target; the therapy is based on brand new discovery of an NK cell dysfunction mechanism in an MASLD disease process, the defect is accurately corrected through genetic engineering, and the immune surveillance function of a body is recovered instead of non-specific anti-inflammation or anti-fibrosis.

The present application relates to the technical field of vaccine preparation, and particularly relates to a tetravalent dengue mRNA vaccine, a liposome nanoparticle and a preparation method and application thereof. The present application provides an mRNA vaccine (mRNA-LNP) encoding a liposome nanoparticle-wrapped EDIII structural protein of four kinds of serum types DENV in series and a polypeptide containing multiple TBT epitopes. After mice are inoculated with the vaccine, specific humoral immunity, cellular immunity with high levels against four kinds of serum types dengue virus are generated, and antibodies with neutralizing effects are also generated. The serum after immunization can protect 3-day-old suckling mice from lethal DENV attack, which indicates that the neutralizing antibodies generated by the vaccine are sufficient to resist the lethal challenge, and the vaccine is an effective vaccine which can be used for long-term research.