432 results about "Ifn gamma" patented technology

The invention discloses a VEGF-resistant and PD-1-resistant difunctional antibody and application thereof, belonging to the technical field of molecular immunology. The VEGF-resistant and PD-1-resistant difunctional antibody contains a light chain an a heavy chain, wherein the light chain has an amino acid sequence as shown in SEQ ID NO.2, and the heavy chain has an amino acid sequence as shown in SEQ ID NO.4 or SEQ ID NO.6. Meanwhile, the invention provides a gene for encoding the difunctional antibody and the application of the difunctional antibody. The difunctional antibody provided by the invention can be combined with PD-1 and VEGF, has very high affinity, can be used for effectively simulating T cells to secrete IL2 and induce T cells to secrete IFN-gamma and can also be used for remarkably inhibiting the growth of tumor of a mouse so as to have a huge potential in application to preparation of anti-cancer drugs.

A method for treating tendonitis and bursitis in a subject involves providing an effective amount of an anti-cytokine agent to a musculo-tendinious structure. Anti-cytokine agents, such as, TNF-α inhibitors, NF-κB inhibitors, IL-1 inhibitors, IL-6 inhibitors, IL-8 inhibitors, IL-12 inhibitors, IL-15 inhibitors, IL-10, Interferon-gamma (IFN-gamma) act to prevent further inflammation initiated by cytokine factors. One embodiment includes, adding with the anti-cytokine agent one or more of an antibiotic or analgesic. Delivery of the anti-cytokine agent may be provided to the affected musculo-tendinious structure by injection, implantation, or a transdermal patch. These agents, individually or in combination directly address the underlying causes of tendonitis, bursitis and associated tendinopathies that result in inflammation and pain.

The invention relates to an ELISpot tuberculosis infection diagnostic kit and the application, which adopts the genetic engineering technology to obtain a CFP10-ESAT6 fusion protein antigen from a mycobacterium tuberculosis by cloning, expression and purification, a tuberculosis specific cell stimulator is used for stimulating the peripheral blood T lymphocyte cell of the detected person to secrete a specific IFN-Gamma, then the invention is detected by ELISpot, and the whole process needs to take two days. The usage of the invention can be used for detecting whether the detected person is infected by mycobacterium tuberculosis by using the naked eye or instrument to determine the result according to the number of the generated purple blue spots, so as to assist the diagnosis of tuberculosis and differential diagnosis.

The invention discloses an efficient multiplication CTL preparation method killing tumors in a targeted mode. The CTL preparation method comprises the following steps: (a) removing CD4+CD25+Treg cells through immunomagnetic bead negative sorting; (b) arranging mixed cells in a serum-free medium for cultivation, and obtaining suspension cells and adherent cells; (c) adding GM-SCF and IL-4 in the adherent cells, culturing the cells for five days; in the sixth day, adding a tumour cell holoantigen, and in the seventh day, adding TNF-alpha and IL-27; (d) transferring the suspension cells to a culture flask wrapped by a CD3 monoclonal antibody and recombinant human fibronectin, adding IFN-gamma, in the second day, adding IL-2, IL-12 and the IL-27, and culturing the mixture till the eighth day to obtain CIK cells; (e) mixing the CIK cells and mature DC cells, and adding the IL-12, IL-7 and an anti-CD 28 monoclonal antibody for cultivation; in the third day, adding an anti-CTLA-4 monoclonal antibody, and then culturing the mixture for four days. According to the efficient multiplication CTL preparation method killing tumors in the targeted mode, efficiency of in-vitro CTL cell proliferation is improved, activity of killing the tumor cells in the targeted mode is improved, transformation of peripheral blood mononuclear cells to the CD4+CD25+Treg cells is inhibited.

The present invention relates to a method and a pharmaceutical composition for treatment of nerve disorders comprising administration of a therapeutically effective dosage of at least two substances selected from the group consisting of TNF inhibitors, IL-1 inhibitors, IL-6 inhibitors, IL-8 inhibitors, FAS inhibitors, FAS ligand inhibitors, and IFN-gamma inhibitors. Preferably, at least one of the substances is a TNF inhibitor.

The invention relates to a method for amplifying cytokine induced kill (CIK) cells and a CIK cell preparation, which belong to the field of in-vitro culture of immune cells. The method concretely adopts the following procedures that: a, lymphocyte cell separation liquid is used for separating out peripheral blood mononuclear cells (PBMC), a culture bag is covered by CD3mAb and CD137mAb in advance, the concentration of the PBMC obtained through separation is regulated to 1*10<6>/ml by a serum-free culture medium, in addition, IFN-gamma is added to obtain the final concentration being 1000 mu/ml, and the materials are transferred to the culture bag to be cultured; b, CD3mAb, CD28mAb and CD137mAb are added after the culture for 24h, in addition, the prepared serum-free culture medium is added, IL-1alpha, IL-2, IL-12 and IL-15 are added into the prepared serum-free culture medium, and obtained CIK cells are collected through centrifugation after the continuous culture for 7 to 21 days; and c, in the culture process of the step b, the cells in the culture bag are counted every three days, in addition, the culture medium is supplemented according to the concentration of the cells, and the CD3mAb, the CD28mAb and the CD137mAb are added to the corresponding concentration every six days, so the CIK cell generative cell times and the cytotoxin activeness are improved.

The invention discloses a kit for assisted diagnosis of tuberculosis, which comprises a specific antibody composition, wherein the specific antibody composition comprises the following five antibodies: (1) IFN-gamma antibody, (2) TNF-alpha antibody, (3) IL-2 antibody, (4) MIG antibody and (5) IP-10 antibody. The kit disclosed by the invention can distinguish a Mycobacterium tuberculosis infected person from a BCG vaccinee. Compared with the ELISPOT tuberculosis diagnosis method, the tuberculosis diagnosis kit based on multi-molecular marker detection has obviously enhanced sensitivity to active tuberculosis (from 72% to 89%), and the positive rate for normal healthy persons is obviously reduced (from 27% to 16%).

The invention discloses a preparing method of high genitality and high cell toxic activity CIK cell, which comprises the following steps: adding gene recombination human cytokine IFN-gamma, CD3 single antibody protein and gene recombination human cytokine IL-2, IL-12 and IL-1 into individual nucleate cell; culturing for 7-24 days under 37 deg. c with CO2 density at 5% and humidity at 100%' getting high genitality and high cell toxic activity CIK cell. This invention increases augmentation multiple and decreases toxic side effect of single cytokine, which can make cell toxic activity maintain 2-3 weeks.

The invention discloses an efficient CIK amplifying method, in particular to a method for cell populations, namely cytokine-induced killer cells utilizing in-vitro cell factors for efficiently inducing mononuclear cell expressions CD3 and CD56 of peripheral blood, wherein the cell populations have killing functions. The cultivation method for the CIK efficiently expressing CD3+CD56 comprises the following steps that the peripheral blood of a healthy person or a patient is collected in a sterile mode, after the peripheral blood is diluted through a saline solution with the same volume, a Ficoll lymphocyte separating medium is used for separating mononuclear cells, in the CIK cell inducing process, CD3 monoclonal antibodies (CD3mAb), CD28 monoclonal antibodies (CD28mAb), interferon-gamma (IFN-gamma), interleukin-2(IL-2) and interleukin 1alpha (IL-1alpha) are added, cultivation is carried out for 13-16 days to obtain cells, a CIK cell preparation is prepared, and flow cytometry detection and microorganism detection are carried out. According to the CIK cultivation method, the CIK number can be increased to be 6*10<9> or over 6*10<9> in two weeks, the cell survival rate can be increased to be 99% or over 99%, and the double-positive proportion of the CD3+CD56+cells reaches 30% or over 30%.

The invention discloses a tuberculosis gene vaccine based on T cell epitopes, wherein a full-length gene, embedded with four T cell epitope polypeptide genes which come from mycobacterium tuberculosis antigen, of mycobacterium tuberculosis heat shock protein is inserted into a vector. The invention also discloses a method for preparing the vaccine, which comprises the following steps: four T cell epitope genes, namely EAST-6189-228, Ag85A369-405, CFP10162-207 and Ag85B420-459 which come from the mycobacterium tuberculosis antigen are inserted into an HSP65 full-length gene. The invention also discloses application of an ECANS tuberculosis gene vaccine. Through the intramuscular injection of the gene vaccine into an immune mouse, the experiment proves that the vaccine can induce a specific antibody which aims at a plurality of tuberculosis antigens to response, can induce stronger tuberculosis specific killing response, can induce Th1 immune response at the same time, secrete high-level IFN gamma, and is a good vaccine for preventing and treating tuberculosis.

The invention relates to a fusion protein MBP-NAP. The amino acid sequence of the fusion protein is shown as SEEQ ID NO.1. Moreover, the invention also discloses a preparation method of the fusion protein MBP-NAP. The method has the characteristics that: the method is easy and convenient to operate, has a mild condition and the like. Th1 cellular immunity of the fusion protein MBP-NAP of the invention is enhanced, so that the secretory volumes of related cell factors IL-2 and IFN gamma are increased remarkably and Th1 immunity is enhanced.

A method of treating an inflammation in a subject in need thereof is disclosed. The method comprises locally or systemically administering to the subject IFN-gamma in an amount so as to achieve an IFN-gamma bulk tissue concentration at a site of inflammation of 1-8,000 units per kilogram body weight, thereby ameliorating the inflammation.

The invention relates to a preparation method of a CAR-T cell targeting B7H3. The preparation method includes first preparing a PBMC cell; then co-transfecting a 293T cell with a shuttle plasmid LV-B7H3 containing the CAR structure, a helper plasmid psPAX2 and an envelope plasmid VSV-G to obtain a packaged B7H3-CAR virus; then taking a PBMC cell, using anti-human CD3 and anti-human CD28 as activators, culturing and activating for 48 hours and adding the B7H3-CAR virus for infection. By means of the preparation scheme, the expression of IFN-gamma in the CAR-T cell is increased, and the cell killing activity is high. The CAR-T cell targeting B7H3 has a killing effect on various solid tumor cells, has high killing activity, is safe and effective, and can be used for immunotherapy of kidney cancer, lung cancer, liver cancer, glioma, ovarian cancer, breast cancer and the like.

The invention relates to a diagnostic kit for tuberculosis and mycobacterium tuberculosis infectors, a preparation method and an application method thereof. By using the linker for encoding 15 amino acid (G4S1) 3, the encoding genes (SEQ.ID.NO.6) of a mycobacterium tuberculosis specific antigen Rv3875 and Rv3874 are connected in series, and then inserted into an E. coli expression vector, and the high-efficiency expression and purification for the fusion protein of Rv3875 and Rv3874 in the E. coli are achieved. The recombinant protein at least comprises 8 T cell epipositions which can be used for cell immunity diagnosis, and a diagnostic kit and diagnostic method for a whole blood IFN-Gamma release analysis method are established by taking the protein as the basis and combining the human IFN-Gamma enzyme-linked immunoassay technology, and can be used for the early, specific diagnosis and screening of tuberculosis and mycobacterium tuberculosis infectors.

The invention discloses an identification and preparation method and application CD106-positive mesenchymal stem cells. The method comprises the following steps: identifying and isolating CD106-positive cells from mononuclear cells isolated from bone marrow, placenta or other tissues through an immunological method, carrying out adherent culture, and collecting adherent CD106-positive cells; or carrying out adherent culture of mononuclear cells isolated from bone marrow, placenta or other tissues, collecting adherent mesenchymal cells, and isolating the CD106-positive mesenchymal stem cells when the count of the obtained cells is large enough. The CD106-positive cells are mesenchymal stem cells with adherent growth, express mesenchymal stem cell-related immunological phenotype, and have osteogenesis, adipogenesis and other differentiation potencies. Compared with CD106-negative mesenchymal stem cells, the CD106-positive cells have higher immunosuppression capacity, can better inhibit proliferation of IFN-gamma (interferon-gamma) secretion of lymphocytes, and can be used for treating graft versus host disease (GVHD) and autoimmune diseases.

The invention discloses a method for detecting a pathogenic microorganism by using antigen-stimulated cellular immune response. In the method, immune cells, which have been exposed to the microorganism, in blood are stimulated by specific antigen of the pathogenic microorganism or a functional segment of the pathogenic microorganism, so a cell factor, such as interferon gamma (IFN-gamma), is produced by the immune cells; and then whether the pathogenic microorganism exists in animals is judged by detecting the amount of the IFN-gamma. In the invention, the IFN-gamma in the blood is detected by a colloidal gold-labeled immunochromatographic assay. In addition, the invention also discloses a colloidal gold immunochromatographic assay test pen for detecting the pathogenic microorganism. By the method, the pathogenic microorganism in an incubation period or early morbidity can be detected quickly and effectively.

The invention discloses porcine parvovirus nanometer alumina gel adjuvant inactivated vaccine which is prepared by mixing inactivated porcine parvovirus virus solution and nanometer alumina gel adjuvant according to the ratio of 1 to 2. According to the invention, safety evaluation to the vaccine is performed on suckling mice and pigs, the results show that after immunization, one suckling mouse dies, no sensitization response is caused on a baby pig, as the same, no obvious full body or partial reaction can be observed, so that the inactivated vaccine is fully proved to be safe and reliable. The porcine parvovirus nanometer alumina gel adjuvant inactivated vaccine set has a CD4 +/CD 8 + relative value in the fourth immunization week, which is obviously higher than that of other vaccine set, the relative value is increased to 2.97 from 2.11, which shows that the vaccine enhances the immune state of cells of mice. The porcine parvovirus nanometer alumina gel adjuvant inactivated vaccine provided by the invention can induce the secretory expression of IFN-Gamma, and the vaccine can remarkably enhance the immune response reaction produced by Th 1 type cells, to induce the Th 1 type cells to secrete IFN- Gamma. Therefore, the immune level of cells can be increased, and a stronger cellular immunity response can be produced.

Here, we describe a sensitive and specific assay and kit for the detection of chemokines having activity that is upregulated by Th-1 cytokines (such IFN-gamma) and chemokines that upregulate the activity of Th-1 cytokines (such as IFN-gamma). In a typical embodiment, detection of the chemokine monokine induced by gamma interferon (MIG) provides a measure of the biological effect of IFN-gamma rather than direct quantitation of IFN-gamma or IFN-gamma secreting cells per se. Upregulation of MIG expression was observed following in vitro activation of PBMC with defined CD8+ T cell epitopes derived from influenza virus, CMV, or EBV, and in all cases this was antigen-specific, genetically restricted and dependent on both CD8+ T cells and IFN-gamma. Responses as assessed by the MIG assay paralleled those detected by conventional IFN-gamma ELISPOT, but the magnitude of response and sensitivity of the MIG assay were superior. Our data validate this novel method for the detection of high as well as low levels of antigen-specific and genetically restricted IFN-gamma activity or MIG.

A method for culturing the protocorm of dendrobium without hormone includes such steps as providing the aseptic test-tube plantlet of dendrobium, inducing, inducing protocorm, naturalizing protocorm, reproducing protocorm, and synthesizing polyose, which can promote the generation of cell interferon (IFN gamma) and tumor necrosis factor (INF alpha).

The invention discloses an application of nano-selenium serving as a CIK (cytokine-induced killer) cell sensitizer. The application of the nano-selenium serving as the CIK cell sensitizer refers to applying the nano-selenium as the CIK cell sensitizer to assist CIK cells. The nano-selenium provided by the invention is at least one of pure nano-selenium, polyethylene glycol modified nano-selenium, polyvinylpyrrolidone modified nano-selenium, chitosan modified nano-selenium, polysaccharide modified nano-selenium, folic acid modified tumor-targeting nano-selenium and transferrin modified tumor-targeting nano-selenium; and the CIK cells are obtained by performing mononuclear cell separation on human peripheral blood, bone marrow or cord blood, and performing induced culture by using a CD-3 antibody and IFN-gamma and IL-2 in vitro. The nano-selenium sensitizer disclosed by the invention has low-price and easily-available raw materials, synthesis and purification steps are strong in operability, and the synthesis scale can be appropriately enlarged by virtue of process optimization, thereby achieving commercialization and application of the medicine.

The invention discloses a method for enhancing mesenchymal stem cell chemotactic capacity and chemotactic factor CCL5 expression, and relates to mesenchymal stem cells. The method comprises the steps of separation of the mesenchymal stem cells and passage of primary culture umbilical cord mesenchymal stem cells; identification of the umbilical cord mesenchymal stem cells; stimulation of TNF-alpha and IFN-gamma to hUC-MSCs; detection of CCL5 expression through Real-Time PCR; detection of CCL5 expression through ELISA. CCL5 expression through MSCs is improved by 924 times, a large amount of natural CCL5 can be obtained, no lipofection transfection is needed, and rapidness and convenience are achieved. Due to simulation of TNF-alpha and IFN-gamma, the transfer ability of the mesenchymal stem cells is improved, it is promoted that the mesenchymal stem cells can more rapidly collect tissue injuries and inflammation portions in vivo, and the functions of tissue repairing and immune modulating are brought into play.