41results about How to "Avoid immune escape" patented technology

The invention belongs to the technical fields of immunology and tumor therapy, and relates to a preparation method and a use of a TCR gene modified CD8+T memory stem cell. The method comprises the following steps: co-incubating a tumor antigen and an immature dendritic cell to obtain a specific tumor antigen-supported mature dendritic cell, co-culturing the specific tumor antigen-supported maturedendritic cell and a CD8+ naive T cell, and adding a stem cell differentiation inhibitor to promote the generation of a CD8+ stem cell-like memory T lymphocyte; separating the CD8+ stem cell-like memory T lymphocyte; and cloning a T cell receptor gene (TCR) for specifically identifying a specific antigenic epitope, and carrying out lentivirus packaging or retroviral vector cotransfection on an autologous CD8+ Tscm cell to in-vitro prepare a CD8+ TCR-Tscm cell specific for different tumor specific antigens. The CD8+ TCR-Tscm cell prepared by the method has the advantages of overcoming of the tumor heterogeneity, high specificity, few adverse reactions, and highly-efficient and lasting tumor preventing and treating effects.

The invention provides a chimeric antigen receptor taking TIGIT and PD-1 as targets, and the chimeric antigen receptor comprises a TIGIT extracellular region and a PD-1 extracellular region; the invention also provides a CAR-T cell containing the chimeric antigen receptor and a preparation method thereof. The CAR-T cell prepared from the chimeric antigen receptor taking the immune checkpoints TIGIT and PD-1 as the targets can promote the proliferation of the CAR-T cell, improve the killing ability of the CAR-T cell, avoid immune escape of tumor cells and improve the effectiveness of CAR-T celltreatment. The Anti-TIGIT-PD-1-CAR-T cells prepared by the method disclosed by the invention is best in proliferation capacity and the most remarkable in effect of killing target cells, and the killing capacity of the Anti-TIGIT-PD-1-CAR-T cells is about three times that of single-target Anti-TIGIT-CAR or Anti-PD1-CAR.

The invention discloses a natural killer cell modified by specific chimeric antigen receptor gene and a preparation method and use thereof. The natural killer cell modified by specific chimeric antigen receptor gene comprises a NY-ESCO-1 specific chimeric antigen receptor gene and a truncated spleen tyrosine kinase Syk gene. The preparation method simultaneously infects the natural killer cell with a slow virus containing the NY-ESCO-1 specific chimeric antigen receptor gene and a slow virus containing the truncated spleen tyrosine kinase Syk gene. By the combined action of the NY-ESCO-1 specific chimeric antigen receptor gene and the truncated spleen tyrosine kinase Syk gene, the natural killer cell modified by the specific chimeric antigen receptor gene improves the targeted tumor cell specific lethality of the natural killer cell, improves the ability of activating downstream signals of the natural killer cell, avoids the escape of tumor immunity, and accordingly enhances the killing activity on the tumor cell.

The invention belongs to the field of tumor biological products, in particular to a chimeric antigen receptor and its gene and a recombinant expression vector, CD19-CD20 dual targeting T cells and anapplication thereof. The chimeric antigen receptor is CD20ScFv-L-CD19ScFv-CD8-CD137-CD3 zeta, which includes CD20ScFv, ligation peptide L, CD19ScFv, a hinge region and a transmembrane region of CD8, an intracellular signal domain of CD137, and an intracellular signal domain of CD3 zeta which are connected in series in order. In the treatment of B cell line hematological malignancy, the chimeric antigen receptor-modified T cells of the present invention can not only specifically recognize tumor cells expressed by a single target of CD19 antigen or CD20 antigen, but also to recognize tumor cellsco-expressed by two targets of CD19 antigen and CD20 antigen, compared with single target CAR T, dual-target CAR T has stronger anti-tumor activity, which avoids immune escape of tumor cells with lowabundance antigen expression, and the risk of recurrence is reduced.

The invention relates to the field of biological medicine, and particularly provides an injection preparation of an anti-PD-L1 monoclonal antibody, which comprises the anti-PD-L1 monoclonal antibody with the protein content of 10-60 mg / ml; a buffer agent having a concentration of 10-60 mM; a stabilizer with the concentration of 100 to 150 mM; an osmotic pressure regulator with the concentration of10-100 mM; and 0.005%-0.05% (w / v) of a surfactant; wherein a pH value of the injection preparation is 5.8 to 6.3. The anti-PD-L1 monoclonal antibody disclosed by the invention has relatively high affinity and good biological activity; meanwhile, different auxiliary materials are selected according to the characteristics of the anti-PD-L1 monoclonal antibody, so that the safety and effectiveness of the monoclonal antibody in the preservation process can be guaranteed, and the physical stability, chemical stability and biological stability of the anti-PD-L1 monoclonal antibody can be maintained.

The invention discloses a CD19 and CD20 targeting double-target chimeric antigen receptor and an expression vector and an application thereof, which belong to the field of tumor immune drugs. The double-target chimeric antigen receptor comprises a signal peptide, a single-chain antibody targeting CD19, a single-chain antibody targeting CD20, a CD8 alpha hinge region, a transmembrane region, a costimulatory factor and an intracellular signal peptide which are connected in sequence. The double-target chimeric antigen receptor can be used for specifically identifying tumor cells expressed by twosingle targets, namely a CD19 antigen and a CD20 antigen, can be used for identifying tumor cells co-expressed by CD19 antigen and CD20 antigen, the double-target CAR-T has stronger anti-tumor activity, and can prevent positive tumor cells expressed by low-abundance antigen from generating immune escape, so that the recurrence risk is reduced.

The invention provides a CD19 and CD30 double-target chimeric antigen receptor and an application thereof. The chimeric antigen receptor comprises an antigen binding structural domain, a transmembranestructural domain and a signal transduction structural domain, wherein the antigen binding structural domain comprises an anti-CD19 single-chain antibody and an anti-CD30 single-chain antibody. The anti-CD19 and anti-CD30 double-target chimeric antigen receptor disclosed by the invention has targeting activity on CD19 positive and / or CD30 positive cells, the T cell for expressing the anti-CD19 and anti-CD30 double-target chimeric antigen receptor has a killing effect on tumor cells with low CD19 antigen expression quantity or no CD19 antigen expression quantity and tumor cells with low CD30 antigen expression quantity or no CD19 antigen expression quantity, so that the immune escape phenomenon is avoided, and the possibility of disease recurrence is reduced.

The invention discloses a chimeric antigen receptor capable of secreting a bispecific antibody as well as an expression vector and an application thereof, which belong to the field of tumor immune drugs. The chimeric antigen receptor comprises a signal peptide, a CD19-targeting single-chain antibody, a lengthened CD8 alpha hinge region, a transmembrane region, a costimulatory factor, an intracellular signal peptide, a P2A connecting peptide, an IL2 signal peptide, a CD20-targeting single-chain antibody, a CD3-targeting single-chain antibody and a tag protein which are connected in sequence. The chimeric antigen receptor is the bispecific antibody capable of specifically targeting CD19 positive tumor cells and secreting targeting CD20 and targeting CD3 at the same time, the non-conductive Tcells can be targeted and activated, so that the tumor antigen CD20 is targeted, and the purpose that the CAR-T cells effectively kill malignant blood tumor cells of a B cell line in vivo is achieved. Meanwhile, the T cells of the chimeric antigen receptor can prevent CD19 positive tumor cells with low abundance expression from generating immune escape, so that the recurrence risk of malignant blood tumors of a B cell line is reduced.

The invention provides a CD19 and CD22 double-target chimeric antigen receptor T cell and application thereof. The chimeric antigen receptor T cell expresses a chimeric antigen receptor which is specifically combined with CD19 and CD22, wherein the chimeric antigen receptor comprises a signal peptide, an antigen binding structural domain, a transmembrane structural domain and a signal transductionstructural domain; the antigen binding structural domain comprises an anti-CD19 single-chain antibody and an anti-CD22 single-chain antibody; and the signal transduction structural domain is 4-1BB, CD3 zeta and TLR2. According to the invention, 4-1BB, CD3zeta and TLR2 which are connected in series are used as a signal transduction structural domain of the chimeric antigen receptor to construct the CD19 and CD22 targeting double-target CAR-T cell, and the CAR-T cell has efficient CD19 and CD22 targeting activity and killing ability, and is beneficial to avoiding immune escape phenomenon.

The invention discloses construction and preparation of double-target chimeric antigen receptor T (CAR-T) of FLT3-NKG2D. CAR of the invention belongs to double-target second-generation CAR, and has the functions of overcoming tumor immune escape and enhancing the specific recognition and killing of tumor cells. By combination of the double-target CAR-T and gilteritinib, the treatment dilemma of refractory acute myeloid leukemia of FLT3<mut+> can be effectively solved, and a new treatment strategy is provided for treatment of acute myeloid leukemia.

The invention provides a CAR-NK92 cell expressing CXCR2, a preparation method and application thereof. Particularly, the invention provides an engineered immune cell expressing chimeric antigen receptors CAR and CXCR2 targeting CD47. CXCR2 can not only help the migration of CD47-targeted CAR-NK cells to tumor cells, but also improve the killing activity of CD47-targeted CAR-NK cells to tumor cells. Particularly, NK cells expressing CAR and CXCR2 containing vSIRP alpha can effectively kill tumor cells, obviously improve the curative effect and reduce recurrence and side effects.

The invention provides a CD19 and CD22 double-target chimeric antigen receptor and application thereof. The chimeric antigen receptor comprises an antigen binding structural domain, a transmembrane structural domain and a signal transduction structural domain; and the antigen binding structural domain comprises a light chain variable region of an anti-CD19 single-chain antibody, a heavy chain variable region of the anti-CD19 single-chain antibody, a light chain variable region of an anti-CD22 single-chain antibody and a heavy chain variable region of the anti-CD22 single-chain antibody. The anti-CD19 and anti-CD22 double-target chimeric antigen receptor disclosed by the invention has targeting activity on CD19 positive and / or CD22 positive cells; the T cells expressing the anti-CD19 and anti-CD22 double-target chimeric antigen receptor has a killing effect on tumor cells with low CD19 antigen expression quantity or no CD19 antigen expression quantity and tumor cells with low CD22 antigen expression quantity or no CD22 antigen expression quantity, so that the immune escape phenomenon is avoided, and the possibility of disease recurrence is reduced.

The invention provides a CD19 and BCMA double-target chimeric antigen receptor and an application thereof. The chimeric antigen receptor comprises an antigen binding structural domain, a transmembranestructural domain and a signal transduction structural domain, wherein the antigen binding structural domain comprises an anti-CD19 single-chain antibody and an anti-BCMA single-chain antibody. The anti-CD19 and BCMA double-target chimeric antigen receptor is small in side effect and high in safety, the constructed CAR-T cell has a killing effect on B cell tumors, and the treatment effect of solid tumors can be remarkably improved.

The invention relates to a PD-1 gene expression silenced engineered immune cell. In particular, the present invention relates to the PD-1 gene expression silenced engineered immune cell that expressesa CAR or an exogenous TCR that targets a tumor cell marker and a fusion protein that targets PD-L1. The experiments show that the engineered immune cell of the present invention is affected by immunosuppression effect of a PD-1 signal pathway, and synergistically activates immune cells by the fusion protein that targets PD-L1, and synergism provides higher immunization cell activity, the tumor cell killing effect is significantly enhanced, the tumor cells expressing the CAR and / or exogenous TCR-targeted antigen and tumor cells expressing the PD-L1 can be simultaneously killed, tumor cells canbe prevented from immune escape, and the immune cell is difficult to target and relapse.

The invention relates to a lymphocyte culture method, and in particular discloses a culturing method of adenovirus gene-modifying tumor specificity cytotoxic T lymphocyte. The method comprises: taking 150-200ml of bleeding of the umbilicus or autologous peripheral blood for single karyocyte separation, separately culturing DC cells by adhering to a wall for 2h, and performing T lymphocyte preculture on the non-adhered cells in an IL-2-contained culture medium; collecting mature DC after the DC cells are mature through induction of iAPA factor adenovirus infection, stimulating the autologous primary T lymphocyte to differentiate into cytotoxic T lymphocyte according to an effect target ratio of DC to T being 1 to 10, and observing and recording multiplication capacity of CTL and detecting the cytotoxicity of the CTL. The limitation of a blood component single sampling machine is solved, collected blood volume is less, burden of a patient is reduced, and iAPA-DC can be observed in vitro to stimulate and differentiate the primary T lymphocyte into tumor specificity cytotoxic T lymphocyte.

The invention belongs to the field of biological medicine, and particularly relates to application of an sPD1 protein and / or an sPD1 gene as an immunologic adjuvant to preparation of a recombinant HPVvaccine. In application, the sPD1 protein contains an amino acid sequence shown as SEQ ID NO:3, or an amino acid sequence with the same function, wherein the amino acid sequence with the same function is obtained after deletion, insertion or replacement of the amino acid sequence shown as SEQ ID NO:3; and the sPD1 gene contains an amino acid sequence shown as SEQ ID NO:1, or an amino acid sequence with the same function, wherein the amino acid sequence with the same function is obtained after deletion, insertion or replacement of the amino acid sequence shown as SEQ ID NO:1. sPD1 serves as the immunologic adjuvant to enhance the ability to stimulate an organism to generate an antibody by the HPV vaccine and induce the organism to generate humoral immune response, stimulate lymphocytes tosecrete IFN-gamma, and stimulate the organism to generate cellular immune response.

The invention relates to an antibody fusion protein targeting Frizzled-7 as well as a preparation method and application of the antibody fusion protein, and belongs to the field of bioengineering. Compared with the prior art, a humanized antibody SHH002-hu1 targeting Fzd7 and an MICA protein are subjected to fusion expression in a genetic engineering mode, Fzd7 targeting treatment and immune activation are combined for the first time through the design, an antibody part of the fusion protein is combined with tumor cells positively expressing Fzd7, and MICA molecules are anchored to the surfaces of the tumor cells. Then, the MICA is specifically combined with an NK cell surface membrane receptor NKG2D, the NK cells are gathered to a tumor focus, and the NK cells are activated to exert a killing effect on the tumor cells, so that immune escape of the tumor in conventional antibody treatment is avoided, and besides, the anti-tumor effect of the antibody is enhanced.

The invention provides a CD19 and CD20 double-target chimeric antigen receptor and application thereof. The chimeric antigen receptor comprises an antigen binding structural domain, a transmembrane structural domain and a signal transduction structural domain, wherein the antigen binding structural domain comprises an anti-CD19 single-chain antibody and an anti-CD20 single-chain antibody. The anti-CD19 and anti-CD20 double-target chimeric antigen receptor has targeting activity on CD19 positive and / or CD20 positive cells, T cells expressing the anti-CD19 and anti-CD20 double-target chimeric antigen receptor have killing effect on tumor cells with low CD19 antigen expression quantity or no CD19 antigen expression quantity and tumor cells with low CD20 antigen expression quantity or no CD20 antigen expression quantity, thereby being beneficial to avoiding an immune escape phenomenon, and reducing the possibility of disease recurrence.

The invention provides a preparing method of a 1-MT-carboxymethyl chitosan drug. According to the preparing method, 1-methyl-DL-tryptophan serves as a raw material, firstly, BOC amino protection is conducted on 1-methyl-DL-tryptophan through di-tert-butyl dicarbonate ester, then 1-methyl-DL-tryptophan and carboxymethyl chitosan are subjected to an amide reaction, through water desorption of BOC groups, the BOS groups are removed, and then the 1-MT-carboxymethyl chitosan drug is obtained. The prepared 1-MT-carboxymethyl chitosan drug inhibits the activity of indoleamine 2,3-dioxygenase with thetumor immune escape function, thus immune escape of indoleamine 2,3-dioxygenase is avoided, and then the immunotherapy effect of the antitumor drug is improved.

The invention discloses a method for preparing super cytokine induced killer (CIK) cells, and belongs to the technical field of medical treatment. By the method, after in vitro amplification tumor killer cells reach the required quantity, cytotoxic T lymphocyte antigen (CTLA4) antibodies are in vitro loaded to cover all CTLA4 molecules on the surfaces of the cells; and all surplus CTLA4 antibodies are washed off after the loading is finished. After the prepared high-activity tumor killer cells are returned by veins, the CTLA4 antibodies achieve the curative effect of the antibody drug, the serious toxic or side effect caused by injecting a large dosage of CTLA4 antibodies from the veins can be avoided, the tumor cells can be continuously killed, and the aims of inhibiting the tumor tissues for long time and even clearing the tumor tissues are finally fulfilled.

The invention relates to the field of biological medicine, and particularly provides an injection preparation of an anti-PD-L1 monoclonal antibody, which comprises the anti-PD-L1 monoclonal antibody with the protein content of 10-60 mg / ml; a buffer agent having a concentration of 10-60 mM; a stabilizer with the concentration of 100 to 150 mM; an osmotic pressure regulator with the concentration of10-100 mM; and 0.005%-0.05% (w / v) of a surfactant; wherein a pH value of the injection preparation is 5.8 to 6.3. The anti-PD-L1 monoclonal antibody disclosed by the invention has relatively high affinity and good biological activity; meanwhile, different auxiliary materials are selected according to the characteristics of the anti-PD-L1 monoclonal antibody, so that the safety and effectiveness of the monoclonal antibody in the preservation process can be guaranteed, and the physical stability, chemical stability and biological stability of the anti-PD-L1 monoclonal antibody can be maintained.

The invention provides a transgenic natural killer cell and a preparation method and application thereof. The provided transgenic natural killer cell comprises a targeting mesothelin antigen-specific chimeric antigen receptor gene and a tyrosine phosphatase SHP gene in a silencing NK cell. The preparation method includes the step of infecting the NK cell with slow viruses containing the targeting mesothelin antigen-specific chimeric antigen receptor gene and slow viruses containing the tyrosine phosphatase SHP gene in the silencing NK cell simultaneously. By the combined action of the targeting mesothelin antigen-specific chimeric antigen receptor gene and the tyrosine phosphatase SHP gene in the silencing NK cell, the transgenic natural killer cell is capable of targeting and identifying tumor cells in a specific and targeting mode to avoid the immune escape of the tumor, and the activity on killing the tumor cells is enhanced.

The invention relates to an mRNA (messenger Ribonucleic Acid) tumor vaccine for improving DCs (Dendritic Cancer) disability in a tumor immune microenvironment as well as a preparation method and application thereof. The mRNA tumor vaccine is constructed from a cation compound loaded with alpha-galactosylceramide (alpha-Galcer), protamine and mRNA, the alpha-Galcer is adsorbed to a phospholipid layer of the compound, and a protamine-HER2 mRNA condensation compound is wrapped in an inner core of the compound. The mRNA tumor vaccine constructed by the invention can be used for immunotherapy through subcutaneous injection or intramuscular injection or nasal mucosa, and can stimulate DCs cells to be mature and secrete cell factors. After mice are immunized, the novel mRNA tumor vaccine can effectively promote T cell proliferation and activation, exert congenital and adaptive immune response, improve the inhibition effect of a tumor immune microenvironment and effectively enhance anti-tumor immune response.

The invention belongs to the field of drugs and health-care food, and discloses a traditional Chinese medicine composition, which contains ganoderma lucidum polysaccharides, grifola frondosa polysaccharides, angelica sinensis oil and cinnamon oil. Experiments show that the prepared traditional Chinese medicine composition is scientifically combined, brings out the best in each other, has no toxic and side effects, not only has nutritional values, but also can inhibit the growth of tumors, remarkably increase the percentage of CD<4+> and CD<8+> cells in tumor stroma lymphocyte compared with single component, enhance the organism immunity function, restore the normal immune monitoring function of the organism, reduce the expression rate of mice tumor tissue VEGF and TGF-beta1 positive cells, help treat tumors, enhance the anti-tumor effect of chemotherapeutic drugs, and improve the organism immunity condition, has the function of restoring the normal immune monitoring of the organism, and can play a role in preventing and / or help treating tumors.

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.

The invention relates to the field of health-caring foods, and especially relates to a Chinese herbal medicine polysaccharide composition and an application thereof. The Chinese herbal medicine is composed of codonopsis pilosula and asparagus cochinchinensis or is composed of codonopsis pilosula polysaccharide and radix asparagi polysaccharide. A test proves that the Chinese herbal medicine composition, medicine or health-caring food is prepared through scientific combinations, wherein the components have synergistic effects and have no side and toxic effects. The composition not only has nutritional value but also can recover normal immune-surveillance function of body, and has prevention and / or auxiliary therapy effects on tumor.