61 results about "Invariant natural killer T-cell" patented technology

The invention provides a natural killer cell, NK-92, modified to express an Fc receptor on the surface of the cell, such as CD16 (FcγRIII-A), or other Fcγ or Fc receptors. The modified NK-92 cell can be further modified to concurrently express an associated accessory signaling protein, such as FcεRI-γ, TCR-ζ, or to concurrently express interleukin-2 (IL-2) or other cytokines. Additional methods are disclosed for various assays, assessments, and therapeutic treatments with the modified NK-92 cells.

A substantially enriched mammalian hematopoietic cell subpopulation is provided, which is characterized by progenitor cell activity for lymphoid lineages, but lacking the potential to differentiate into myeloid and erythroid lineages. Methods are provided for the isolation and culture of this common lymphoid progenitor cell (CLP). The cell enrichment methods employ reagents that specifically recognize CDw127 (IL-7 receptor α); CD117 (c-kit) protein, in conjunction with other markers expressed on lineage committed cells. The murine cells are also characterized as expressing low levels of sca-1 (Ly-6E and Ly-6A). The CLPs are predominantly cycling, blast cells. These cells give rise to B cells, T cells and natural killer cells, as evidenced by their growth and differentiation in vitro and in vivo.

The present invention relates to a method for inducing and expanding natural killer cells derived from peripheral blood mononuclear cells, which comprises co-culturing, as feeder cells, irradiated Jurkat cells and irradiated Epstein-Barr virus transformed lymphocyte continuous line (EBV-LCL) cells in the presence of cytokines, along with peripheral blood mononuclear cells. According to the present invention, a large quantity of natural killer cells can be induced and proliferated from a small quantity of peripheral blood mononuclear cells even without the use of high-cost equipment or various kinds of expensive cytokines, thereby making it possible to significantly improve the efficiency and efficacy of the prevention and treatment of cancer using the natural killer cells.

Provided herein are sphingolipid compounds that are useful for activating natural killer T cells. Also provided are methods for treating or preventing a disease or disorder that is treatable by activating the immune system by stimulating natural killer T cells. The compounds are therefore useful for treating or reducing the likelihood of occurrence of an immune diseases and disorders, such as autoimmune diseases or disorders. The compounds may also be used for treating or reducing the likelihood of occurrence of a microbial infection or for treating or reducing the likelihood of occurrence of a cancer in a subject by administering the sphingolipid compounds described herein.

Modified glycolipid compounds are provided. Also disclosed are methods for activating an NKT cell, methods of stimulating an immune response in a subject, and methods suitable for labeling NKT cells.

The invention discloses a method for jointly preparing CAR-NK (chimeric antigen receptor-natural killer) cells and CAR-NKT (natural killer T) cells. The method includes steps of 1), adding peripheral blood mononuclear cells into cell culture media, pre-activating NK cells and NKT cells in the peripheral blood mononuclear cells and selectively activating and amplifying the NK cells and the NKT cells in an in-vitro manner; 2), transducing mixed cells obtained at the step 1) by the aid of lentiviral vectors with gene sequences of chimeric antigen receptors. The method has the advantages that the CAR-NK cells and the CAR-NKT cells can be prepared from NKs and NKTs of healthy donors in a standardized and batched manner; the purity of the NK cells and the NKT cells can respectively reach 60% and 30% at least without purification, and accordingly preliminary purification can be omitted; the in-vivo activation and amplification degrees and the in-vivo existence time of the CAR-NK cells and the CAR-NKT cells which are jointly prepared by the aid of the method can be controlled by means of clinical medication.

The present invention relates to an improved growth method for natural killer cells (NK cells). More specifically, the present invention relates to a growth method for natural killer cells, which comprises a step of culturing natural killer cells in a medium containing anti-CD3 antibodies and interleukin proteins in the presence of peripheral blood leukocytes. The present invention provides an innovative growth method for natural killer cells, which can obtain a large amount of natural killer cells by remarkably increasing the growth rate compared with conventional growth methods for natural killer cells.

Human Valpha24<+> natural killer T cells are expanded by culturing a mononuclear cell fraction obtainable from human peripheral blood in which hemopoietic stem cells are mobilized by granulocyte colony-stimulating factor, in the presence of a cytokine, such as interleukin 2, effecting proliferation and/or activation of lymphocytes and alpha-glycosylceramide. A cell fraction comprising human Valpha24<+> natural killer T cells expanded by the method, is useful as a cancer-treating agent.

The invention belongs to the technical field of genetic engineering antibodies, and particularly relates to a preparation and application of a bispecific antibody targeting a CD24 (cluster of differentiation 24) and activating NK cells (natural killer cells). According to the preparation and application of the bispecific antibody, an anti-CD24 humanized chimeric antibody cG7 and an MICA molecule capable of recruiting and activating the activity of the NK cells are connected through flexible peptide (Gly4Ser) and are stably transferred to a CHO-s eukaryotic expression system, and a stable high-expression cell strain is selected for propagation and expression of cG7-MICA; the bispecific antibody cG7-MICA utilizes the function of remodeling the MICA by utilizing the targeting effect of the cG7 to selectively display the MICA on the surface of a CD24+ tumor cell, the immune surveillance effect of the NK cells is induced, in addition, the NK cells are further activated through the effect ofan Fc part, and the antibody-dependent cell-mediated cytotoxicity (ADCC) is achieved; and through experimental verification, the immunocompetence of killing the CD24+ hepatocellular carcinoma cells is achieved in vitro and vivo.

NK cells and NK cell lines are modified to increase cytotoxicity, wherein the cells and compositions thereof have a use in the treatment of cancer. Production of modified NK cells and NK cell lines is via genetic modification to remove checkpoint inhibitory receptor expression and/or add mutant (variant) TRAIL ligand expression.

The invention discloses an invariant natural killer T (iNKT) cell expressing a targeted GPC3 chimeric antigen receptor and preparation and application for the iNKT cell. The chimeric antigen receptor contains a hinge region, a transmembrane region and an intracellular signal region of a single chain antibody GPC3-ScFv, CD8 which recognize a C tail end epitope of GPC3, wherein the hinge region, the transmembrane region and the intracellular signal region are connected according to series structure domains. Preparation of the iNKT cell modified by the chimeric antigen receptor comprises the following steps that a chimeric antigen receptor pRRL-gc33-28BBz is built, and the iNKT cell is infected; and after special amplification in vitro, the GPC3-targetted iNKT cell is obtained. According to the iNKT cell expressing the targeted GPC3 chimeric antigen receptor and preparation and application for the iNKT cell, the nucleic acids encoding the chimeric antigen receptor protein, a plasmid containing the nucleic acids, a virus containing the plasmid and the transgenic iNKT lymphocyte transduced by the virus can be effectively applied to tumor immunotherapy.

The invention provides a culture method for monkshood polysaccharide-induced nature killer T cell (Nature Killer T cell, NKT) proliferation. The method comprises the characteristics as follows: human peripheral blood, marrow or a mononuclear cell with an umbilical cord blood source are continuously cultivated by traditional Chinese medicine monkshood polysaccharide and recombinant human interleukin 2, a lot of nature kill T cells are obtained, NK<1.1+> and CD<8+> are expressed, interferon gamma is secreted, the ratio in lymphocyte at the 14th day is more than 16.15%, the order of magnitude can be up to over 5*10<9>, and the nature kill T cell have effective anti-tumor and anti-virus functions. A clinical application of the NKT cell obtained by the method is also provided by the invention.

The invention relates to the field of biotechnology and medical science, in particular to a fusion protein for NKT (natural killer T) cell culture, an encoding gene and an application. The fusion protein is used for preparing NKT cells which are high in proliferating activity and killing activity. The fusion protein ICAM1-FN (intercellular adhesion molecule-1-fibronectin) can effectively stimulate proliferation of lymphocytes in the culture process of the NKT cells, the proportion of CD3/CD56 double-positive cells is improved, so that the in-vitro amplification efficiency of peripheral blood mononuclear cells is twice that of a traditional method, the proportion of CD3/CD56 double-positive cells is 2-10 times that of the traditional method, tumor killing activity of the NKT cells is enhanced, and the tumor killing activity of the prepared method of the NKT cells is improved by 15%-35% as compared with the traditional method.

The invention provides a natural killer T cell culture medium and a multiplication culture method for natural killer T cells and relates to the technical field of cell culture. The natural killer T cell culture medium, disclosed by the invention, is mainly composed of a serum-free medium, autologous plasma and a cell factor IL-2. According to the multiplication culture method for natural killer T cells by using the cell culture medium, human-derived peripheral blood mononuclear cells are induced to release a danger signal by virtue of the antibody binding cell factor IL-2, and further the natural killer T cells are activated. According to the method, high-quality natural killer T cells can be obtained by massive high-efficiency multiplication in a short period on premise of ensuring the security. The potential security risk caused by culture of heterogenous animal serum or tumor cells in the conventional natural killer T cell culture method and the problems that the conventional natural killer T cell culture method is low in multiplication times and low in purity and is difficult to use on a large scale are effectively solved.

The invention discloses a method for efficient in-vitro amplification and culture of natural killer T cells with high killing capacity. The method comprises the following steps: a, taking fresh human peripheral blood and obtaining mononuclear cells with a gradient centrifugation method; b, pre-stimulating mononuclear cells in an in vitro medium; c, expanding culture of peripheral blood derived mononuclear cells, detecting the proportion of various cells, and judging amplification efficiency and activity of the target NK/NKT cells; d, detecting the proportion of NK/NKT cells to the cell population after culture for 3-4 weeks, and detecting cell killing efficiency; e, further purifying the NK/NKT cells. The method is simple to operate, the NK cell population rich in NKT cells with high killing activity is obtained and contains no impurity cells, immunogenicity caused by the impurity cells is avoided, and side effects in clinical application are lower. Besides, the method has the advantages that materials are easily available, the cells are suitable for large-scale culture, damage to the body is small and the like, and is more suitable for adjuvant treatment of tumor patients.

Provided herein are placental perfusate, placental perfusate cells, and placenta-derived intermediate natural killer (PINK) cells, and combinations thereof. Also provided herein are compositions comprising the same, and methods of using placental perfusate, placental perfusate cells, and placenta-derived intermediate natural killer cells, and combinations thereof, to suppress the growth or proliferation of tumor cells, cancer cells, and the like, and to treat individuals having tumor cells. In a preferred embodiment, the composition comprises: solated CD56+, CD16- natural killer cells.

The disclosure concerns a method for cancer treatment by in vivo priming and activation of natural killer cells for achieving tumor cell lysis. The method includes introducing into a patient a priming tumor cell preparation (PTCP) derived from a first tumor cell line, which is irradiated to inactivate the first tumor cells or a membrane preparation thereof, the first tumor cells having known priming ligands on the membrane surface thereof. The patient's rest NK cells are contacted by the PTCP in vivo, resulting in primed NK cells, which are characterized by upregulation of CD69, shedding of CD16, or a combination of CD69+ and CD16−. These primed NK cells then contact second tumor cells, the cancer, and are configured to lyse and kill the second tumor cells.

The manipulation of human dendritic cells (DCs) to induce potent anti-tumor immunity remains an essential subject of study. Here we report that the overexpression of CD1d in human DCs can enhance the priming of naïve CD8+ T cells against tumor antigen. We showed that CD1d can be overexpressed in the human DCs using baculoviral vector carrying the CD1d gene. This CD1d-overexpression is functional as demonstrated by the increased expansion of invariant natural killer T (iNKT) cells while using these modified DCs to present α-galactosylceramide (α-GC). Pulsed with tumor antigenic peptide, these CD1d-overexpressing human DCs showed enhanced capability to prime naïve CD8+ T cells. CD1d-overexpressing human DCs also induced a pro-inflammatory cytokine profile that may favor the priming. Moreover, this CD1d-overexpression strategy can be extrapolated to monocyte-derived human DCs. Therefore, our study suggest that overexpression of CD1d in human DCs may provide a novel strategy to enhance DC immunogenicity and the possible translation into human cancer immunotherapy.

The invention discloses a functionality detection and evaluation method for liver disease NK (natural killer) cells, comprising the steps of (1) passing single cells through a flow cytometer, and using a combination of different anti-cytokine or NK cell surface receptor antibodies and cellular surface or intracellular specific markers to detect corresponding cytokine secretion or surface receptorlevels; (2) using a big data bio-statistical analytic technique to interpret the contribution, action and influence of each immune factor upon whole immunity, and finally NK cell overall immunity interpretation for the action of the immune factors upon immunity. The invention also provides a functionality detection kit for liver disease NK cells, comprising the surface molecular antibodies: CD56,NK-IFN-gamma, NK-TNF-alpha, NK-G2D, NK-p46, NK-p30, NK-p44, NK-KIRs, NK-LAIRs, NK-Perforin, NK-GrazymeB, NK-G2A, NK-PD-1, and NK-Tim3. The functionality detection and evaluation method and functionality detection kit herein can effectively detect overall functionality of NK cells.