111 results about "Gamma delta T cell" patented technology

The invention belongs to the field of medical biology engineering, and particularly relates to a method for effectively multiplying gamma delta T cells by stimulating peripheral blood in vitro and application of the method. The method comprises the step of using feeder cells, an OKT3 (ornithine ketoacid transaminase) antibody, interleukin-2 and zoledronic acid. The feeder cells are formed by specifically inserting CD64, CD86 and CD137L genes in a target site of a genome of the feeder cells. After the zoledronic acid and the nterleukin-2 are used for increasing the proportion of the gamma delta T cells of the peripheral blood, protein products of genes, the OKT3 antibody and the interleukin-2 act in a combined manner, and the gamma delta T cells can be stimulated so that a large amount of gamma delta T cells can be multiplied. The multiplied gamma delta T cells can be used for killing tumor cells which are pretreated by the zoledronic acid, or the tumor cells can be directly killed by modifying and expressing chimeric antigen receptors (CAR) via a genetic engineering means. The gamma delta T cells which are obtained by the method have complete anti-tumor cytotoxicity, and can kill solid tumor cells and non-solid tumor cells.

The invention relates to a method for culturing gamma-delta-T cells, and in particular relates to a method for in-vitro amplification of gamma-delta-T cells, wherein the method comprises the following operating steps of: pre-coating a T75 culture bottle by a TCR-gamma-delta resisting antibody and CD28McAb for later use use; isolating the peripheral blood mononuclear cell (PBMC) of a patient; regulating the PBMC concentration to 1*10<6> 6/ml by a serum-free culture medium which contains 5% of autologous plasma, and transferring PBMC cell suspension into the T75 culture bottle; adding an initial culture medium which contains proper concentrations of Zoledronat, HSP70, 1L-2, 1L-7 and 1L-15; culturing in a saturated humid environment containing 5% of CO2 at 37 DEG C; depending on growth situation of the cell, changing the culture medium every 2-3days, to control the cell concentration at about 2.5*10<6>/ml; meanwhile, compensating full doses of Zoledronat, HSP70, 1L-2, 1L-7 and 1L-15; and continuously culturing for 12-16days, to obtain a great amount of gamma-delta-T cells which are comparatively high in purity.

The invention discloses an expansion method of various lymphocyte subpopulations and application of the expansion method to preparation of an adoptive immunotherapy medicine for cancers. The expansion method comprises the following steps: S1, adding IFN gamma and zoledronic acid into a culture medium of PBMC collected in the blood of a tumor patient, and re-adding an OKT3 antibody and recombinant human interleukin-2 after culture to stimulate PBMC so as to finish preliminary expansion; S2, after finish of preliminary expansion, mixing immune lymphocytes with feeder layer cells, and after adding zoledronic acid, an OKT3 antibody and recombinant human interleukin-2, continually performing expansion. By adoption of the expansion method, after twice expansion, the sum of the immune cells can be 10,000 to 80,000 times by expansion, the expanded immune cells include alpha beta T cells, gamma delta T cells, NKT cells and NK cells, and tumor cells can be directly killed, or a cellular immunotherapy drug is prepared to kill the tumor cells.

The invention relates to a composition for stimulating and inducing a single aryocyte to be amplified to a gamma delta T cell. The composition is prepared from zoledronic acid, anti-human CD3Ab, anti-human CD28Ab, IL-15, IL-21 and IL-2. The invention also provides a method for the in-vitro mass amplification of gamma deltaT cells by utilizing the composition, and particularly relates to a method for in-vitro inducing the mass amplification of gamma deltaT cells by utilizing PBMCs. The zoledronic acid is used for induction and activation, the anti-human CD3Ab, the anti-human CD28Ab, IL-15, IL-21 and IL-2 are used for inducing and activating the proliferation, and various amplification factor combinations to jointly stimulate and induce the amplification of gamma deltaT cells, and the obtained gamma deltaT cell has the characteristics of large quantity, high purity, high cell toxicity and the like and has good clinical application value.

The invention discloses a method for co-culturing, inducing and amplifying gamma delta T cells and NK cells. The gamma delta T cells and the NK cells are obtained in a co-culture manner; peripheral blood is collected to separate and screen mononuclear cells and stimulate and induce the mononuclear cells to induce the gamma delta T cells and the NK cells, the gamma delta T cells and the NK cells are subjected to activation amplification culture to amplify a gamma delta T cell and NK cell composite cell substance having characteristics of large number, high amplification multiple, strong cytotoxicity and the like so as to form the co-culture cell composition through compounding, and the co-culture cell composition has good clinical application value. The co-culture induced amplification method improves the cell culture efficiency, greatly reduces the culture cost, reduces the difficulty of a culture technology, simplifies the clinical treatment process, reduces the treatment cost, and simplifies the treatment means.

This invention relates to the expansion of non-haematopoietic tissue-resident γδ T cells in vitro by culturing lymphocytes obtained from non-haematopoietic tissue of humans or non-human animals in the presence of interleukin-2 (IL-2) and/or interleukin-15 (IL-15) and the absence of TCR activation or co-stimulation signals, without any direct contact with stromal or epithelial cells. Methods of non-haematopoietic tissue-resident γδ T cell expansion are provided, as well as populations of non-haematopoietic tissue-resident γδ T cells and uses thereof.

The present invention concerns a method of inducing IL-2-free proliferation of γδ T cells using a combination of a γδ T cell activator and IL-33 for use in therapy of infection, cancer, autoimmunity as well as other diseases.

The invention relates to the technical field of cell culture, in particular to a building method of NK (natural killer) cell and gamma delta T cell co-culture capable of realizing the in vitro commonmultiplication culture on NK cells and gamma delta T cells. The method mainly comprises the following steps of S1, reagent selection; S2, performing PBMC culture by a culture medium for 3 days; S3, supplementing the culture medium; maintaining the concentration; performing culture for 4 days; S4, after the seventh day, removing supernatant through centrifugation; transferring the cells into a T25cell culture bottle; then, replacing an EX culture medium; S5, after the tenth day, supplementing the culture medium for maintaining the concentration; S6, after the twelfth day, continuously supplementing the culture medium for maintaining the concentration; S7, after the fourteenth day, detecting the NK cell and gamma delta T cell proportion and the cell total number. The co-culture building method has the advantages that two kinds of anti-tumor immune cells with similar properties are cultured in one step; the cell culture efficiency is improved; the culture cost can be greatly reduced through being compared with that of independent culture of various immune cells for obtaining NK cells and gamma delta T cells; the culture technical difficulty is also reduced.

The invention provides application of a novel gamma delta T cell subpopulation to preparation of a kit for predicting the curative effect and prognosis of AML (acute myeloid leukemia). The inventor based on the invention discovers the expression condition of the novel gamma delta T cell subpopulation in peripheral blood of patients suffering from AML is associated with the curative effect and theprognosis of the patients suffering from AML for the first time. When the expression ratio of the novel PD1+Foxp3+gamma delta T cell subpopulation is high, the possibility of bad clinical curative effect of the patients suffering from AML is high. The expression ratio of the novel gamma delta T cell subpopulation has important guide significance in prognosis judgment of the patients suffering fromAML and formulation of a clinical treatment scheme. More base research data can be provided for individual treatment of the patients suffering from AML, and wide application prospect in clinical curative effect and prognosis evaluation of the patients suffering from AML is achieved.

The invention discloses a method for preparing a CAR-T cell from gamma delta T cell derived from umbilical cord blood, the CAR-T cell and an application and also relates to nucleic acid coding a chimeric antigen receptor and a recombinant expression vector expressing the chimeric antigen receptor. The umbilical cord blood is used as a source of the gamma delta T cell, the gamma delta T cell is separated from the umbilical cord blood, after efficient amplification, the T cell is transduced by a lentiviral vector loaded with the chimeric antigen receptor, and the CAR-T cell can be obtained, so that positive tumor cells or other infectious cells which can efficiently or specifically kill related antigens are obtained. The CAR-T cell will play roles in tumor cell therapy and immunotherapy.

The invention relates to the field of immune medicine, in particular to a cell culture method which comprises the following steps: co-culturing an inactivated artificial antigen presenting cell with high expression of mIL15 and CD137L and PBMC in a culture medium, wherein the culture medium comprises a basic culture medium and additional components, the additional components comprise IL2 with the concentration of 100 IU/mL to 1200 IU/mL and zoledronic acid with the concentration of 3 mu M to 7 mu M. The ratio of NK cells to gamma delta T cells cultured by the method is high, and the cells have a more excellent anti-tumor effect.

The invention relates to the medical field and discloses a culture method for a gamma delta T cell. The culture method comprises the steps of carrying out induced culture on a mononuclear cell containing IL-15, OKT-3 and IL-2 in a serum-free culture medium until the gamma delta T cell is induced, then adding a tumor cell lysis solution, continuing to carry out culture and multiplication, then adding TNF-alpha, continuing to carry out culture to promote the maturity and further multiplication of gamma delta T cell, and culturing for 14 days to obtain the gamma delta T cell. Cell factors of an induced culture medium are regulated in the earlier stage, and stimulation and sensitization are carried out by virtue of the tumor cell lysis solution and TNF-alpha in the middle and later stage of the culture, so that the amplification multiple of the cell is increased, the killing activity is enhanced, and the secretion amounts of IFN-gamma and IL-4 are increased.

Provided herein is a method of expanding clinically-relevant quantities of polyclonal γδ T cells that have anti-tumor, anti-viral, and anti-bacterial reactivity. Polyclonal γδ T cells can target a variety of tumors, including solid tumors as well as other conditions, such as viral and bacterial infections.

The invention provides a gamma delta-T cell exosome used for tumor immunotherapy. The gamma delta-T cell exosome is loaded with miR-138. The exosome (gamma delta TDE) derived from gamma delta-T cellscarries exogenous miR-138 for realizing direct targeted recognition and killing on tumor cells, meanwhile, PD-1/PD-L1 is inhibited, the antitumor immunity activity of T lymphocytes/NK cells is enhanced, and the effect of inhibiting tumor growth/invasion/metastasis is achieved through direct and indirect paths.

An activated antigen-presenting cell capable of inducing an immunocompetent cell including a disease antigen-specific CD8+CTL and/or a gamma delta T cell in vivo and/or in vitro with good efficiency; a pharmaceutical comprising the activated antigen-presenting cell; a therapeutic/prophylactic method using the activated antigen-presenting cell; a method for induction of an immunocompetent cell including a disease antigen-specific CD8+CTL and/or a gamma delta T cell which is induced by using the activated antigen-presenting cell; an immunocompetent cell induced by the method; a pharmaceutical comprising the immunocompetent cell; and a therapeutic/prophylactic method using the immunocompetent cell. An antigen-presenting cell is sensitized with a disease antigen and also co-sensitized with bisphosphonate, thereby producing an increased number of a disease antigen-specific CD8+CTL and/or a gamma delta T cell at a higher ratio compared to a method without co-sensitization with bisphosphonate.