100 results about "Dc vaccine" patented technology

The invention belongs to preparation of biological cell formulations, and in particular relates to a preparation method of dendritic cell (DC) vaccine loaded with autologous tumor associated holoantigen. The preparation method comprises the following steps: preparing the autologous tumor associated holoantigen, collecting and separately culturing DCs, impacting the DCs by the autologous tumor associated holoantigen, maturing the DCs and preparing autologous tumor antigen specific DC vaccine. The invention solves the problems in the prior art that the immunogenicity of tumor antigen is not strong enough, the antigen target spots are incomplete, the tumor antigens of most tumor patients are difficult to acquire, and the like. The DC vaccine provided by the invention has the advantages of effectively inducing tumor antigen specific cytotoxic T lymphocyte (CTL) in vitro and in vivo, efficiently generating specific cytotoxicity on tumors, having high overall effective rate and no obvious toxic side effects in clinical application, and the like.

The invention discloses a preparation method for the dendritic cell (DC) of an umbilical cord blood source and a dendritic cell (DC) vaccine, which relates to a preparation method for the dendritic cell. According to the method, various cell factors are adopted to induce DC obtained by umbilical cord blood separation, and then the DC is stimulated by a tumor specific antigen so as to improve the specific antigen presentation capability of the DC; and a stem cell growth factor and Flt3-L are added into a cell culture medium so as to effectively accelerate a hematopoietic cell in the umbilical cord blood to induce and proliferate to an immune cell. The DC vaccine prepared with the method has the specific antigen presentation capability, can be combined with a CIK (cytokine induced killer) cell to mutually treat the malignant tumor when being used as a tumor immunotherapy product, and is used as an important adjuvant therapy after operations and chemoradiotherapy. Recurrence and metastasis after the operations can be effectively prevented, and toxic and side effects caused by the chemoradiotherapy on patients are lowered so as to improve the treatment effect.

The invention belongs to the cellular immunity field, and concretely relates to a preparation method of dendritic cell vaccine. The method comprises the following steps: separating cord blood to obtain cord blood monocyte; screening DC cell from the cord blood monocyte, culturing and performing amplification, then performing single epitope multiple antigen peptide (SEA-MVP) for loading, and then incubating overnight to obtain the DC vaccine. The method has the beneficial effect that the single epitope multiple antigen peptide (SEA-MVP) is used for loading DC cells, the single epitope multiple antigen peptide (SEA-MVP) is obtained by in vitro synthesis, the composition is clear and single, the purity is high; alkaline amino acid is taken as a frame for connecting the single epitope, so that the orientation of the connected epitope peptide has consistency and the epitope peptide is easily recognized and presented by DC cells, the sensitivity and singularity are strong, the loading efficiency is high, and immune tolerance is not generated; compared with the current method, the antineoplastic cell capability of the dendritic cell vaccine for activating the CD4+ and CD8+ cells is increased by more than ten thousand times.

The invention discloses a method for preparing a tumor-specific DC vaccine by applying CD34+ cells in umbilical cord blood. The method comprises (1) a step of preparing autologous tumor-related holoantigen; (2) a step of obtaining the umbilical cord blood; (3) a step of obtaining mononuclear cells derived from the umbilical cord blood; (4) a step of purifying CD34+ cells in the mononuclear cells derived from the umbilical cord blood; (5) performing induction culture for a precursor DC; (6) a step of performing amplification and culture of an immature DC; and (7) a step of preparing the DC vaccine.

The invention discloses a preparation method of a high-efficiency killer cell preparation adopting an immunodetection point dual-block CTL (cytotoxic lymphocyte). The preparation method comprises steps as follows: step 1), collecting mononuclear cells in peripheral blood; step 2), performing DC (dendritic cell) separation and T cell separation on the separated mononuclear cells with an adherence method; step 3), adding adhering DCs to a serum-free culture medium containing GM-CSF (granulocyte-macrophage colony-stimulating factor) and IL-4 (interleukin-4) for preparation of a DC vaccine; step 4), adding suspending T cells to CD3 and IL-2 for T cell culture and amplification; step 5), adding antigen-supported DCs to the amplified T cells for preparation of the CTL, and adding PD-1 (Programmed death 1) and CTLA-4 (cytotoxic t lymphocyte associated antigen-4) antibodies for preparation of the cell preparation with the dual-block CTL effect. According to the novel technology and the novel method provided by the invention, an immune brake effect is effectively regulated and T cell depletion is retarded by blocking inhibitory molecules on the tumor specific CTL surfaces, so that the tumor cytotoxicity of CTL effector cells is substantially improved.

The invention provides a method for producing a dendritic cell (DC), which involves the step of culturing a DC precursor cell in the presence of two or more cytokines; a DC produced by the method; and use of the DC. The method can produce a large quantity of a DC precursor cell which is highly capable of being differentiated into a DC. The method can also produce a large quantity of a DC from a small quantity of a DC precursor cell. Therefore, it becomes possible to readily increase the number of DCs to be administered in the anti-tumor immunotherapy or the treatment of an infectious disease utilizing a DC, resulting in the enhancement of the efficacy of a DC vaccine.

The invention provides an HPV (human papillomavirus) peptide / DC (dendritic cell) mixed vaccine prepared in the presence of a TLR (toll-like receptor) agonist. The mixed vaccine comprises one part of HPV11E77-15, one part of mouse bone marrow-derived dendritic cells which are cultured in vitro and one part of TLR9 agonist as an adjuvant, wherein the dendritic cells are extracted from mouse bone marrow and further cultured in vitro, the co-incubation with one section of HPV11E7CTL (cytotoxic T lymphocyte) epitope peptide with strongest immunogenicity is firstly carried out, the co-incubation with the TLR ligand CpG and the like is further respectively carried out for promoting the maturation of the DC, the degree of maturation can be confirmed by detecting the change of a marker on the surface of the DC after incubation, and the DC vaccine which is simulated to mature can be used for immunizing mice. The TLR ligand and the HPV11E7 peptide are utilized jointly to promote the degree of maturation of the DC, the level of TNF-alpha (tumor necrosis factor-alpha) and IFN-gamma (immunoreactive fibronectin-gamma) of factors of secretory cells of T cells can be particularly improved after combination of the CpG, and the HPV peptide / DC mixed vaccine can be used for preventing and treating genital warts and cervical cancer.

The invention relates to an antigen sensitized dendritic cell (DC) preparing method. On one hand, a culture medium for preparing antigen sensitized DCs is provided and is composed of a basal culture medium and other ingredients, the basal culture medium and the other ingredients can be conveniently mixed into the culture medium for culturing the DCs before using, the other ingredients comprise pharmaceutical grade human albumin, recombinant human insulin and the like, and the culture medium has the advantages of being definite in ingredient, free of ingredients of animal origin and the like; on the other hand, the invention provides a method for preparing the antigen sensitized DCs through the culture medium, and the method includes the steps that a single karyocyte is cultured and induced through the culture medium to obtain immaturate DC cells; strong immunogenicity tumor antigens are prepared through high hydrostatic pressure, astragalus polysaccharide is united to serve as an immunologic adjuvant, antigen sensitization is conducted on the immaturate DCs, and the antigen sensitized DCs are obtained. The preparing method has the advantages of being safe, efficient, easy to implement and the like. The DC vaccines for specific therapy can be provided for clinic through the method.

The present invention provides methods for producing DCs, which comprise the step of culturing DC precursor cells in the presence of multiple cytokines, dendritic cells produced thereby, and uses thereof. The methods of the present invention enable production of large quantities of DC precursors with a high ability to differentiate into DCs. The present invention enables one to obtain large quantities of DCs from a small number of DC precursor cells, and therefore makes it easier to increase the number of DCs for administration in DC-based anti-tumor immunotherapy, treatment of infection, and such. Thus, an enhancement is expected for the effect of DC vaccines.

The invention provides an efficient tumor antigen loaded DC vaccine and a method for induced proliferation of tumor antigen specific CTL by the same. The DC vaccine is prepared by steps: (1) tumor antigen preparation; (2) acquisition, separation and efficient culture and proliferation of DC cells; (3) adoption of a tumor antigen and an activator for sensitization and activation of the DC cells; (4) DC cell surface PD-L1 molecule closing. The invention further provides a sequential blood drawing method for in-vitro induced proliferation of CTL by the DC vaccine and closing of surface PD-1 molecules of CTL subjected to in-vitro induced proliferation. Problems of low immunogenicity of existing tumor antigens, incompleteness of antigen targets, difficulty in acquisition of tumor antigens of patients and insufficiency in DC and CTL in-vitro proliferation are solved. The DC vaccine is effective in in-vitro and in-vivo induction of tumor antigen specific CTL and capable of efficiently generating specific killing effects on tumors without evident toxic and side effects, and a promising clinical application prospect is achieved.

The invention discloses a tumor associated antigen XAGE-1b short peptide and application thereof. The sequence of the short peptide is one of SEQ ID NO:2-SEQ ID NO:15. CTL established by induction of XAGE-1b antigen peptide cannot generate an immunologic response to testis cells and only kills tumor cells. CTL clone obtained by induction has a favorable specific tumor cell killing effect; meanwhile, during a CTL establishment process, the inventor discovers that the screened XAGE-1b antigen peptide has appropriate appetency with HLA on DC cells and is capable of effectively stimulating and inducing production of specific CTLs, which indicates that the XAGE-1b antigen peptide has favorable polypeptide vaccine and DC vaccine potentials. By virtue of cloning the specific TCR genes of the XAGE-1b antigen peptide, a virus carrier is established, peripheral circulation CD8+T cells are transduced, TCR gene modified T cell (TCR-T) clone is obtained and also has a similar specific tumor cell killing effect with the parent-generation CTL, which indicates that the TCR gene modified T cell clone has favorable clinical conversion and practical application prospects.

The invention discloses a tumor composite antigen, a dendritic cell multivalent vaccine and application of the tumor composite antigen and the dendritic cell multivalent vaccine. Dendritic cells of a patient are stimulated in vitro, multiple tumor cell lysates with super immunogenicity aiming at different EBV related tumors are loaded, mature dendritic cells are formed under induction of multiple cell factors and specific agonists, a complete DC vaccine with corresponding cancer antigens is formed, the DC vaccine is transfused back to a human body to activate an immune system, and the EBV related tumors are subjected to tumor immunogenicity. Natural immunity (such as induction of NK cells) is stimulated, lymphocytes are stimulated to generate acquired immune response, cytotoxic T cells are generated to kill cancer cells, and the cancer cells are accurately killed together; compared with radiotherapy and chemotherapy, the composition is particularly safe and almost has no side effect; and the dendritic cell vaccine has a preparation cycle of about 1 week, and is short in time and low in cost.