181 results about "Natural killer T cell" patented technology

The invention relates to an engineered CD20 targeting NKT cell and its preparation method and application. The cell is an NKT cell modified by a chimeric antigen receptor CD20ScFv-CD8-CD137-CD3zeta. An amino acid sequence of the chimeric antigen receptor is as shown in SEQID NO.1 in a sequence table. The preparation method comprises the following steps: firstly constructing a chimeric antigen receptor pWPT-CD20ScFv-CD8-CD137-CD3zeta, carrying out infection of the NKT cell, inducing in vitro and carrying out amplification culture so as to obtain the engineered CD20 targeting NKT cell. During advanced CD20 positive malignant tumors, the engineered CD20 targeting NKT cell can be used to obviously prolong survival time of immune cells in a patient's body, enhance the immune cells' targeting recognition capability of tumor antigen and reinforce the killing activity of tumor cells.

The invention discloses a method for culturing natural killer (NK) and/or natural killer T (NKT) cells. The method comprises the following step: inoculating isolated NK and/or NKT cells into a culture system A for culture to obtain propagated NK and/or NKT cells, wherein the culture system A consists of a buffer solution containing CD3 antibody and/or Retronectin and inducing factors. Experiments prove that peripheral blood mononuclear cells (PBMC) extracted from peripheral blood are separated and enriched through magnetic beads, high-purity CD56+ cells are obtained, two proteins, namely Retronectin and CD3mAb are added into an in-vitro culture system for joint stimulation, and IL-2 and IL-5 factors are used for assisting in induction, so that a culture method capable of obtaining massive NK and NKT cells with high killing activity is established.

The invention relates to the technical field of cell cryopreservation liquids, and in particular relates to a cryopreservation liquid for cultured NK cells and a preparation method of the cryopreservation liquid. The cryopreservation liquid for the cultured NK cells comprises tea polyphenol, a grape seed extract, vitamin C and autoserum. By virtue of adopting the autoserum, pollution of exogenous animal viruses and introduction of exogenous proteins are avoided, and the safety is high; by virtue of adding the tea polyphenol, the grape seed extract (procyanidine) and the vitamin C, cell oxidization can be effectively suppressed, the activity of the cells can be retained, and no harm to a human body is generated; meanwhile, the state of the cells recovered from cryopreservation is basically close to the state before cryopreservation, and the cryopreservation effect is good.

This invention relates to the in vivo expansion of NKT cells by their exposure to mature dendritic cells expressing α-galactosyl ceramide and to methods of use thereof in modulating immune responses, such as anti-cancer responses, and enhancing memory responses.

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 provides a natural killer cell culture substrate and an amplification culture method for natural killer cells, and relates to the technical field of cell culture. The natural killer cell culture substrate is mainly composed of a serum-free culture medium, autologous plasma and cytokines; according to the amplification culture method for the natural killer cells with the natural killer cell culture substrate, the IL-2, IL-12, IL-15, IL-21 cytokines in the cell culture substrate are combined through Lymactin-NK antibodies to induce mononuclear cells of human peripheral blood to release dangerous signals, the natural killer cells are activated, and the killing activity of the cells is enhanced. According to the method, a large quantity of high-quality natural killer cells can be obtained in a short period through efficient amplification, and the problems are effectively solved that in existing culture methods for the natural killer cells, the production process is complex, the production cost is relatively high, and natural killer cells are low in amplification multiple, not high in purity and difficult to produce in a large-scale mode.

Provided are an iPS cell derived from a somatic cell such as an NKT cell, having the α-chain region of the T cell antigen receptor gene rearranged to uniform Vα-Jα in an NKT cell receptor-specific way, NKT cells differentiated from the iPS cell, a method of creating the same, and an immune cell therapy agent prepared using cells differentiated from the iPS cell. Also provided are an iPS cell having TCRα rearranged to NKT-TCR (NKT-iPS cell), obtained by contacting a somatic cell, such as an NKT cell, having the α-chain region of the T cell antigen receptor gene rearranged to uniform Vα-Jα in an NKT cell receptor-specific way, with nuclear reprogramming factors, isolated NKT cells obtained by differentiating the iPS cell ex vivo (iPS-NKT cell), a method of generating CD4/CD8-double positive NKT cells (DP-NKT cells) and mature NKT cells from NKT-iPS cells by altering the combination of feeder cells and/or cytokines, a method of expanding the iPS-NKT cells, and an NKT cell cytotherapy agent comprising NKT cells activated with α-galactosyl ceramide (α-GalCer), or iPS-NKT cells, and α-GalCer in combination.

The invention relates to the field of cells and discloses an addition agent of NKT cell induction culture and a method of induction culture. The addition agent of the NKT cell induction comprises growth factors IL-21, IL-7 and OKT-3. The growth factors IL-21, IL-7 and OKT-3 has a synergistic effect with the recognized growth factors IL-2 and IL-15, the multiplication of the NKT cells can be better induce and promoted, the mass multiplication of the NKT cells can be promoted, and the addition agent has a great killing effect to tumor cells. The method for inducing and culturing the NKT cells is simple in operation, capable of promoting the mass multiplication of the NKT cells and suitable for the scale production of the NKT cells.

The invention provides a recombinant lentiviral expression vector, and belongs to the technical field of cell engineering. The recombinant lentiviral expression vector comprises a 21mb gene, a 41BBL gene and an MICA gene which are sequentially connected in series; and the 21mb gene is obtained by sequentially connecting a signal peptide gene, an IL21 gene, a CD8 Hinge gene and a CD8 transmembranegene in series. After the provided recombinant lentiviral expression vector transfects a host cell, the host cell expresses the MICA and the 41BBL, and the IL21 is fused and expressed (but not secreted in a culture environment) on a cytomembrane of the host cell. After the provided recombinant lentiviral expression vector transfects the host cell, an obtained reconstitution cell promotes the highexpression of the NKG2D, avoids excessive activation of an NK cell and prolongs the proliferation time of the NK cell in the process of induction culture of the NK cell.

The invention discloses a chimeric antigen receptor as well as a gene and a recombinant expression vector, an engineered CD30-targeted NKT cell and an application thereof. The chimeric antigen receptor is CD30ScFv-CD8-CD137-CD3 zeta, which comprises CD30ScFv, a hinge region and a transmembrane region of CD8, an intracellular signal structural domain of CD137, and an intracellular signal structural domain of CD3 zeta which are connected in series connection. The chimeric antigen receptor CD30ScFv-CD8-CD137-CD3 zeta modified NKT cell is used for treating CD30-positive Hodgkin lymphoma and non-Hodgkin's lymphoma; the chimeric antigen receptor has a good and specific killing activity for lymphoma cells, and has a certain treatment effects for CD30-positive Hodgkin lymphoma patients who have been treated for many times without obvious curative effects (such as CD30 monoclonal antibody combined with cytotoxin medicines or radio isotope therapy and the like).

The invention relates to a preparation method used for high efficiency amplification of natural killer (NK) cells. The preparation method comprises following steps: peripheral blood is collected, andsingle karyocytes are separated; the karyocytes are introduced into a culture bottle treated via coating with CD16 monoclonal antibody and HER2 monoclonal antibody for culturing; IL-2, IL-15, IL-21, and inactivated autologous serum are introduced into a culture medium at the same time; the cells are transformed into a culture bottle free of coating treatment, liquid supply is carried out every 2 to 3 days; and at last the obtained NK cells are collected. According to the preparation method, no heterogenous serum or trophoblastic cell is adopted, operation process is simple, high NK cell yieldis achieved, and the clinical application value is high.

The present invention relates to a method for the treatment of immune-related or immune-mediated disorders in a mammalian subject in need of such treatment. This method comprises the step of manipulating the NKT cell population in said subject by suitable means, said manipulation of the NKT cell population resulting in modulation of the Th1/Th2 balance toward anti-inflammatory cytokine producing cells. Manipulation of the NKT cell population may be performed either by depletion of said cells by a suitable means or alternatively by ex vivo education of the NKT cells, such that the educated NKT cells have the capability to modulate the Th1/Th2 balance toward anti-inflammatory cytokine producing cells. The invention further relates to pharmaceutical compositions for the treatment of immune-related or immune-mediated disorders in a mammalian subject. These compositions comprising as an effective ingredient an ex vivo educated NKT cell. The invention further provides for an ex vivo educated NKT cell and uses thereof in the treatment of immune-related or immune-mediated disorders.

Provided is a method for preparing natural killer cell with high efficiency using irradiated peripheral blood mononuclear cells, more particularly to a method for proliferating highly activated NK cells using a combination of irradiated peripheral blood mononuclear cells (PBMCs) and a CD16 antibody and an anti-cancer cell therapeutic composition containing the natural killer cells (NK cells) prepared thereby. Further provided is a method for large-scale proliferation of activated NK cells with high efficiency using a combination of irradiated peripheral blood mononuclear cells (PBMCs) and a CD16 antibody without the use of cancer cells or genetically modified feeder cells having safety issues as feeder cells. The highly purified and highly cytotoxic NK cells proliferated in large quantities can be used as an active ingredient of a cancer immunotherapeutic composition.

The invention provides an immune cell culture medium and an immune cell culture method, as well as an application of ginsenoside. The immune cell culture medium comprises serum-free basal culture medium, ginsenoside and cytokines, wherein the weight content of ginsenoside in the serum-free basal culture medium is 4mg/mL-10mg/mL. The immune cell culture medium provided by invention can enhance the activity and the multiplication capacity of the immune cell. The experimental result shows that after CIK cells are cultured by the immune cell culture medium by 14 days, the proliferation rate is 650-750 times, the survival rate is 97.78-98.73%, the ratio of the cells with the CD3+CD56+ effects is 37.4%-38.2%; after NKT cells are cultured by the immune cell culture medium by 14 days, the proliferation rate is 480-550 times, the survival rate is 96.73-98.78%, and the ratio of the cells with the summation effects of CD3+CD56+ and CD161+ is 42.6%-45.2%.

The invention belongs to the field of cytobiology, and discloses a culturing method for NKT cells. The culturing method for the NKT cells comprises the steps that mononuclear cells are inoculated into an X-VIVO15 serum-free culture medium, a hydroxypropyl methyl cellulose solution, 1L-2, 1L-15, 1L-21 and CD3 monoclonal antibodies are added, and then the NKT cells are cultured. According to the culturing method, the normal-temperature gelling effect of hydroxypropyl methyl cellulose is utilized, the hydroxypropyl methyl cellulose solution is added, a three-dimensional structure of a culture solution is built in the culturing initial stage of the NKT cells, therefore, the NKT cells proliferate in the three-dimensional space, the space is bigger, and the area of the contacted culture solution is sufficient. Experimental results show that by culturing the NKT cells through the culturing method for the NKT cells, the cells are good in state and high in proliferation speed and proliferation multiple, the NKT effect cells with the higher purity can be obtained, and the killing effect of the obtained NKT cells is obviously higher than that of NKT cells obtained through a conventional culturing method.

The invention discloses a method for jointly preparing CAR-VGamma9VDelta2T cells and CAR-NKT cells, comprising the steps of 1) adding peripheral blood mononuclear cells to a cell medium, pre-activating VGamma9VDelta2T cells and NKT cells therein, and amplifying the VGamma9VDelta2T cells and NKT cells by selective in-vitro activation; 2) transducing the mixed cells of step 1) by using a lentiviral vector carrying chimeric antigen receptor's gene sequence. It is possible to carry out standard and batch preparation herein by using GammaDeltaT of a healthy donor; the purities of the VGamma9VDelta2T cells and NKT cells in the application can reach 60% and 30% and above respectively without purification, and therefore pre-purification can be omitted; in-vivo activation proliferation level and existence time of CAR-VGamma9VDelta2T cells and CAR-NKT cells jointly prepared by using the method can be controlled through clinical medication.

The present invention describes a gene encoding a single polypeptide encoding three components of HLA-E cell surface expression. When the gene is introducted into porcine cells it fold properly and confers protection against human NK cell-mediated killing. Additionally, this invention provides a method to inhibit CTL-mediated killing of cells.

The present invention relates to a method for treating recurrent tumor metastases following liver resection that includes administration of an effective amount of an agonist of A_2A adenosine receptors (ARs).

The present invention relates to a method for the treatment of immune-related disorders in a mammalian subject in need of such treatment. This method comprises the step of manipulating the NK T cell population in said subject by suitable means, said manipulation of the NK T cell population resulting in modulation of the Th1/Th2 balance toward anti-inflammatory cytokine producing cells. Manipulation of the NK T cell population may be performed either by depletion of said cells by a suitable means or alternatively by ex vivo education of the NK T cells, such that the educated NK T cells have the capability to modulate the Th1/Th2 balance toward anti-inflammatory cytokine producing cells. The invention further relates to pharmaceutical compositions for the treatment of immune-related disorders in a mammalian subject. These compositions comprising as an effective ingredient an ex vivo educated NK T cell. The invention further provides for an ex vivo educated NK T cell and uses thereof in the treatment of immune-related disorders.