160 results about "Immune escape" patented technology

The invention provides a preparation method of bispecific chimeric antigen receptor gene modified natural killer cells, comprising the features: constructing a bispecific chimeric antigen receptor gene containing specific-binding signaling lymphocyte activation molecule family member 7 and fibronectin mutants, recombining the bispecific chimeric antigen receptor gene to a viral vector, transfecting with human natural killer cells, and highly expressing the bispecific chimeric antigen receptor gene, thus specifically binding tumor cells that express the signaling lymphocyte activation molecule family member 7 and fibronectin mutants, inhibiting expression of natural killer cell inhibitory receptors, and preventing immune escape of tumor cells; meanwhile, activating a first signal and a co-stimulatory signal to trigger toxic activity of the tumor cells, and great anti-tumor killing toxicity is shown in in-vivo and in-vitro experiments; the invention also provides a kit for the preparation of autologous natural killer cells through the above method, and with the kit, it is possible to highly express the bispecific chimeric antigen receptor gene and trigger great anti-tumor effect.

The invention belongs to the field of tumor biological products, and particularly relates to an chimeric antigen receptor, a gene of the chimeric antigen receptor, the recombinant expression vector ofthe chimeric antigen receptor, a CD22-CD19 double-targeting T cell and application of the T cell. The chimeric antigen receptor is CD22ScFv-L-CD19ScFv-CD8-CD137-CD3zeta and comprises CD22ScFv, connecting peptide L, CD19ScFv, a hinge region and a transmembrane region of CD8, an intracellular signal structural domain of CD137 and an intracellular signal structural domain of the CD3zeta which are connected in series. According to the chimeric antigen receptor, the gene of the chimeric antigen receptor, the recombinant expression vector of the chimeric antigen receptor, the CD22-CD19 double-targeting T cell and the application of the T cell, when B-cell line hematological malignant tumors are treated, the T cell modified by the chimeric antigen receptor not only can specifically recognize a tumor cell which is only expressed by a single target spot of a CD19 antigen or a CD22 antigen, but also can recognize tumor cells co-expressed by the two target spots of the CD19 antigen and the CD22antigen, compared with the single target spot CAR T, the double target spots CAR T have stronger anti-tumor activity, so that the immune escape of the tumor cells expressed by the low-abundance antigen is avoided, and therefore the recurrence risk is reduced.

The invention discloses a preparation method of a human skin stem cell factor nano-liposome-exosome complex. According to the method, the exosome and the liposome are compounded, and through the synergistic effect between the exosome and the nano-liposome coated with the cytokine, the obtained compound system has good biocompatibility and immune escape performance, is free of cytotoxicity, not prone to being degraded by macrophages, good in stability and capable of continuously playing a role in vivo.

The invention provides an antibody targeting SARS-CoV-2 and a preparation method and application thereof. The antibody comprises VH and VL, wherein the VH comprises the following CDRs: VH CDR1, VH CDR2 and VH CDR3 with amino acid sequences as shown in SEQ ID NO: 1, 2 and 3; the VL comprises the following CDRs: VL CDR1, VL CDR2 and VL CDR3 with amino acid sequences as shown in SEQ ID NO: 4, 5 and 6. The antibody can be highly compatible and specifically combined with RBD of S proteins of SARS-CoV-2, inhibits combination of the RBD protein and receptor ACE2 protein, efficiently inhibits SARS-CoV-2 from infecting cells, and has very good neutralizing activity on potential pseudoviruses of immune escape mutation, so that the antibody can be effectively applied to diagnosis, prevention and treatment of SARS-CoV-2 viruses and related diseases.

A dendric cell tumor vaccine carrying withered heat shock tumor cells for preparing antineoplastic medicine and decreasing immune escape is prepared through treating tumor cells by heat shock method, inducing the wither of tumor cells, preparing dendric cells, and carrying the withered heat shock tumor cells.

The invention provides construction and application of a fourth-generation chimeric antigen receptor T (CAR-T) cell (expressing IL-7 and CCL19) specific to Nectin-4 dual targets on the surface of a malignant tumor. Two spatial epitopes of a Nectin-4 antigen are taken as target points, and the constructed fourth-generation CAR-T is used for treating and expressing a malignant solid tumor of the Nectin-4 antigen to solve the immune escape problem of the CAR-T during treatment of the solid tumor; and the constructed fourth-generation CAR-T can enhance proliferative capacity and sustained survivability, so that the solid tumor is effectively treated, and a new strategy is provided for effective prevention and treatment of postoperative recurrence / metastasis of the solid tumor.

The invention relates to a preparation method of a cytotoxic T lymphocyte (CTL) targeting to multiple KRAS mutant antigen epitopes. The method comprises the steps: cloning a new artificial antigen coding sequence formed by series combination of 7 main KRAS mutant antigen epitope sequences into a lentivirus expression vector, transfecting DC, stimulating the proliferation and differentiation of activated CD8+T cells, and making the finally prepared cytotoxic T lymphocyte (CTL) having the specificity of targeting to seven main KRAS mutant antigen epitopes. The prepared KRAS mutant antigen specific cytotoxic T lymphocyte (CTL) has the advantages of high targeting specificity, no side effects and not easily causing of immune escape, and has great application prospects in treatment of KRAS mutant tumors.

The invention provides a targeting antitumor T cell. The targeting antitumor T cell comprises an IL13Ralpha2-targeting chimeric antigen receptor CAR-IL13Ralpha2 and / or an HER2-targeting chimeric antigen receptor CAR-HER2, wherein the CAR-IL13Ralpha2 comprises amino acid sequences of an IL13Ralpha2-targeting single chain antibody, an extracellular hinge region, a transmembrane region and an intracellular signal region, which are sequentially connected from an amino terminal to a carboxyl terminal, and the CAR-HER2 comprises amino acid sequences of an HER2-targeting single chain antibody, an extracellular hinge region, a transmembrane region and an intracellular signal region, which are sequentially connected from the amino terminal to the carboxyl terminal. The targeting antitumor T cell can efficiently recognize and kill tumor cells of surface IL13Ralpha2 and / or HER2 positive antigens, can effectively overcome immune escape of tumor cells, and has lasting tumor killing effects. The invention also provides a preparation method and an application of the targeting antitumor T cell.

The invention discloses siRNA capable of knocking down human PD-1, a recombinant expression CAR-T vector as well as a construction method and an application thereof. The PD-1 knocking-down siRNA expression cassette and an siRNA expression product thereof can be applied to CAR-T therapy of multiple myeloma (MM) for eliminating or relieving an immune escape mechanism of tumors and can be also used for inhibiting the immune escape mechanism in the CAR-T therapy of such tumors as pancreatic cancer, brain glioma, myeloma and the like.

The invention relates to the technical field of biological medicine, in particular to an anti-African swine fever virus and anti-CD double-target pig-derived antibody, a preparing method and application. The prepared anti-African swine fever virus and anti-CD pig-derived double-target compound antibody has two antigen combination sites which are combined with African swine fever viruses and immunecells respectively to generate the bridging effect and realize the two major functions; on one hand, the antigen proteins which cause immunosuppression and immune escape are inactivated, the immunosuppression and immune escape are eliminated, and the African swine fever viruses entering a body are killed; on the other hand, the functions of T cells are activated, the immune system is restored, the cell-mediated immunity is started, and virus expansion is prevented; meanwhile, infected cells are split and dissolved, phagocytes are activated to clear away the viruses and the infected cells, andthe aim of preventing and treating the African swine fever viruses is achieved. By means of the compound antibody, a biological medicine and a biological safety feed additive which eliminate the infection of the African swine fever viruses and prevent and treat the African swine fever can be prepared.

The invention discloses a double-target chimeric antigen receptor targeting an NKG2D ligand and CD19 as well as 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 NKG2D extracellular region of a targeted NKG2D ligand, a connecting fragment, a single-chain antibody of the targeted CD19, a lengthened 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 two single targets, namely a targeted NKG2D ligand or a CD19 antigen; in addition, tumor cells co-expressed by the targeting NKG2D ligand and the CD19 antigen can be recognized, the double-target CAR-T has stronger anti-tumor activity, immune escape of positive tumor cells expressed by low-abundance antigen can be avoided, andthus the recurrence risk is reduced.

The invention belongs to the field of tumour basic research, and relates to a method used separating and purifying microglia cells from glioma specimens. The method comprises following steps: a cell suspension is prepared from tumour tissues via mechanical method combined with enzymatic digestion method; the cell suspension is subjected to density gradient centrifugation so as to obtain the microglia cells of CD11B+ with a relatively high purity via enrichment and selection; and the microglia cells with a relatively high purity are obtained via further separation using CD11B+ magnetic beads. Wherein, Percoll separating medium is composed of 70% of Percoll, 30% of Percoll, and HBSS. The purity of the microglia cells obtained via percoll density gradient centrifugation enrichment can be more than 95%, and it is confirmed by immunofluorescence assay and phagocytosis assay that functions of the microglia cells are excellent. The method is simple and convenient for operation, is low in cost, is stable in quality, and is capable of providing basis for further research on immunologic functions microglia cells in brain glioma and effects of microglia cells in immune escape of tumour.

The invention relates to an engineered immune cell targeting CD19 and CD22 and application of the engineered immune cell, and particularly provides a nucleic acid construct having a structure of P1-X-P2-Y (formula I), the definition of each element is as described in the specification. The engineered immune cell can recognize both CD19 and CD22 antigens, and the expression levels of two CARs targeting the CD19 and CD22 are similar in the engineered immune cell, thereby reducing immune escape risks caused by the down-regulation or loss of antigen presentation during recognition of a single-target CAR-T cell in the therapy process, and greatly reducing the difficulty of industrial application of the engineered immune cell.

The invention provides a chimeric antigen receptor T cell and application thereof. The chimeric antigen receptor T cell expresses chimeric antigen receptors targeting CD19 and PD-L1 at the same time.The chimeric antigen receptor T cell expresses a CD19 CAR structure and a PD-L1 CAR structure at the same time, and after PD-L1 CAR is combined with PD-L1 expressed by a tumor cell, an activation signal can be transmitted intracellularly. Therefore, the chimeric antigen receptor T cell not only can recognize the tumor cells expressing CD19 in a targeted manner, but also can relieve immunosuppression of PD-1 expressed by the tumor cells on the T cells, avoid an immune escape mechanism of the tumor cells and relieve the problem of drug resistance of CAR-T treatment; and a new idea is provided for CAR-T treatment and adoptive feedback immune cell treatment.

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 relates to a pharmaceutical composition of cytomembrane bionic titanium dioxide nanoparticles. The pharmaceutical composition can effectively achieve immune escape and long circulation,and promote apoptosis of tumor cells through enhanced acoustic therapy combined with NO chemotherapy. According to the technical scheme, the surfaces of hollow mesoporous titanium dioxide nanoparticles are covalently modified by hemoglobin through an amidation reaction, then physically loaded with an NO donor type chemotherapy drug RRx-001, and finally coextruded with cytomembrane vesicle obtainedthrough pyrolysis treatment of a hypotonic lysis solution and homogenate with cells, and the pharmaceutical composition of the cytomembrane bionic titanium dioxide nanoparticles is obtained. The pharmaceutical composition of the cytomembrane bionic titanium dioxide nanoparticles has the advantages that a preparation method is stable and reliable, the production cost is low, the prepared pharmaceutical composition of the cytomembrane bionic titanium dioxide nanoparticles can play a role of the enhanced acoustic therapy combined with NO chemotherapy in the preparation of antitumor drugs, enhances the antitumor effect, has immune escape and long circulation and other functions, is good in biocompatibility, and is innovation of tumor therapy drugs.

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 provides a construction method of a mouse model with conditional over-expression of HPV E7 gene at H11 locus. The construction method of the mouse model with the conditional over-expression of the HPV E7 gene at the H11 locus comprises the following steps: designing and obtaining sgRNA; (2) preparing a Cas9 / sgRNA mixture; (3) constructing a targeting vector; (4) co-injecting the targeting vector and the Cas9 / sgRNA mixture into fertilized eggs of a mouse so as to subsequently obtain F0 generation mice; and (5) obtaining F1 generation mice. The construction method of the mouse model with the conditional over-expression of the HPV E7 gene at the H11 locus solves the problem that "nude mice are not suitable for studying role of HPV E7 in HPV E7-mediated processes of tumor microenvironment immune regulation, immune escape and the like for the nude mice lack competent immune systems" in the prior art. The mouse model provided by the invention has a competent immune system, so that the mouse model is suitable for all researches on interaction between the HPV E7 and the immune system.

The invention relates to the construction of targeted NK cells used for tumor immunotherapy, and specifically relates to a preparation method for a membrane surface engineered NK cell, a pharmaceutical composition and an application. A targeting vector is adopted to introduce an azide group or DBCO on the surface of a NK cell, and polypeptide targeting PD-L1 can be coupled to the surface of the NKcell through copper-free click chemical reaction, so that a membrane surface engineered NK cell can be constructed. An application of the membrane surface engineered NK cell in treating PD-L1 positive tumor including head and neck squamous cell carcinoma, non-small cell lung cancer and triple negative breast cancer is also disclosed. Compared with the prior art, the preparation method has the following advantages: the targeting of the NK cell can be strengthened, so that potential hazards caused by viral vector transfection can be eliminated; the problems of CAR-T cell individualization and complex manufacturing processes can be solved; and the tumor infiltration capabilities and targeted killing activity of the NK cell can be enhanced, and immune escape mediated by a PD-1 / PD-L1 pathway can be effectively blocked, so that the membrane surface engineered NK cell has potential clinical application prospets.

The invention discloses culturing, screening and amplifying techniques for hair follicle stem cells for clinic treatment level cell therapy. The bottlenecks are always research hotspots and difficulties in the biological stem cell field; currently, by carrying out separation and purification by virtue of remarkable adhering properties of a basement membrane, the hair follicle stem cells are obtained by virtue of three-dimensional culture and amplification measures. By depending on amplification anchorage-dependent cells, a three-dimensional high-simulation in-vivo extracellular matrix system is provided for screening and separating the levels of target cells. The method is a novel culture technique for culture in vitro of biological cells of stem cells of adults (embryo skin) in a high-simulation in-vivo adhesion environment. Skin seed hair follicle cells of targeted tissue engineering cultured through adhesion screening in the environment of a high-simulation substrate have high abdication capacity and multiplication capacity and can form a differentiated cortex structure in vitro. By screening immunogenic cells, immune escape is safely achieved, the transplant time is prolonged, the clinic application of tissue engineering is increased, the healing of wounds is promoted, and a solution with absolute advantages for the clinic treatment level cell therapy is provided for diseases of skin.

The invention discloses a nano photothermal therapy drug and a preparation method thereof. The nano photothermal therapy drug is composed of a nano-drug particle inner core and an outer cancer cell membrane, wherein the nano-drug system is composed of a heat shock protein inhibitor and polydopamine. The medicine can play roles in cancer cell membrane immune escape and tumor site self-targeting, photothermal therapy of polydopamine and sensitization low-temperature photothermal therapy of a heat shock protein inhibitor, carry out strike therapy for the thermal resistance of tumor cells, quicklyrelease the heat shock protein inhibitor and reduce the hepatotoxicity of the heat shock protein inhibitor, has excellent photothermal imaging and fluorescence imaging characteristics, and can be used for cancer targeted imaging and phototherapy; and high-accumulation and low-dose HI formed at self-targeting tumor sites can achieve an ideal effect, and possible toxicity to healthy organs due to excessive use is avoided.

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 application of a cancer cell membrane bionic nano reactor AGZ@CM in preparation of anti-cancer drugs. The efficient synergistic treatment method for tumors comprises the following step: performing one-pot synthesis on a nano reactor AQ4N / GOx@ZIF-8 by virtue of an amino-acid-induced bionic method to realize rapid encapsulation and ultrahigh loading efficiency of glucose oxidase and keep the natural biological activity of the glucose oxidase. The bionic nano-reactor AGZ@CM prepared by coating the surface of AGZ with a targeting cell membrane coating has homotype targetingand immune escape functions; after a cascade reaction in which the ZIF-8 carrier is decomposed in the presence of acid to release a drug, GOx mediated glucose oxidation is performed and a prodrug AQ4N is activated, synergistic treatment of hunger therapy and chemotherapy is realized. In-vitro and in-vivo research results show that the synergistic strategy of hunger therapy and chemotherapy can improve the treatment effect, and the cell membrane coating endows the nano-reactor with strong targeting ability; the bionic intelligent cascade reaction nanoreactor disclosed by the invention has theclinical potential of treating tumors.

Provided is a use of miRNA-214 inhibitor in inhibiting regulatory T cells (Treg cells). The microRNA-214 (miRNA-214) can promote Treg cells, and can assist tumours in immune escape. The experiments demonstrate that the inhibition of miRNA-214 can inhibit the growth of Treg cells, thus inhibiting the growth of tumours. Therefore, miRNA-214 can be used for developing an anti-tumour drug or a drug inhibiting immune response hyperactivity.

The invention belongs to the technical field of immunotherapy, and particularly relates to a dual-CAR (chimeric antigen receptor) structure targeting bispecific sites, an expression vector containing the dual-CAR structure of the bispecific sites, an immune cell and an application. The dual-CAR structure is a ScFv1-hinge-transmembrane-41BB intracellular signal-CD3 [zeta]-self-cleavage polypeptide-ScFv2-hinge-transmembrane-CD28 intracellular signal-CD3 [zeta] or a ScFv1-hinge- transmembrane-CD28 intracellular signal-CD3 [zeta]-self-cleavage polypeptide-ScFv2-hinge-transmembrane-41BB intracellular signal-CD3 [zeta]. The construction of the dual-CAR structure overcomes the technical effect defect of dual-target CAR in the prior art, two different tumor antigens can be targeted at the same time, the killing rate of the tumor cells is increased, and the probability of immune escape of the tumor cells is reduced.

The invention discloses a scoring method for quantifying a TIME infiltration mode; the score for quantifying the TIME infiltration mode is called as a TIME index (TI), the TI is an excellent prognosisprediction index in hepatocellular carcinoma and pancreas carcinoma, and the performance of the TI in predicting immunotherapy response is superior to that of other 11 biomarkers. The results can strengthen the understanding of the immune microenvironment and guide more effective personalized treatment, and the TIME1 is characterized by immune cell depletion and cell proliferation and correspondsto immunodeficiency types; tIME2 is characterized by being in an immunosuppression state, and a potential immune escape mechanism of each phenotype is emphasized corresponding to immunosuppression: TIME1 is characterized by tumor infiltration leukocyte deficiency and tumor antigen presentation capability defects; the TIME2 has more immunosuppressive cells; the TIME3 is rich in immunosuppressive cytokines and immune checkpoint molecules.

The invention relates to an immunomodulatory specific chimeric antigen receptor cell as well as a preparation method and application thereof. According to the invention, a recombinant vector of a chimeric antigen receptor of immune checkpoint regulation (PD1 or TIGIT) combined with specifically targeting human NKG2DL, and is used for modifying immune response cells; the obtained novel functionalized immune response cell can effectively target and attack various tumors, and a preparation for treating malignant tumors is prepared. Engineering immune cells modified by the chimeric antigen receptors of immune checkpoint regulation (PD1 or TIGIT) combined with specifically targeting human NKG2DL is adopted, inactivation of immune cells induced by inhibition factors is inhibited, immune escape of tumor cells is prevented, and antitumor activity can be obviously improved.