50 results about "NK cell proliferation" patented technology

Methods of ex-vivo culture of natural killer (NK) cells are provided and, more particularly, methods for enhancing propagation and/or functionality of NK cells by treating the cells with a nicotinamide or other nicotinamide moiety in combination with cytokines driving NK cell proliferation. Also envisioned are compositions comprising cultured NK cells and therapeutic uses thereof.

The invention is directed to purified and isolated novel ULBP polypeptides, the nucleic acids encoding such polypeptides, processes for production of recombinant forms of such polypeptides, antibodies generated against these polypeptides, fragmented peptides derived from these polypeptides, and the uses of the above. ULBP polypeptide can be found on the surface of human B cell lymphomas. Mammalian forms of ULBP polypeptide in isolated or purified forms are provided. In addition, isolated nucleic acids encoding ULBP polypeptides and expression vectors comprising a cDNA encoding ULBP polypeptides are provided. The ULBP polypeptides can be isolated or synthesized and used to prepare antibodies, and in particular monoclonal antibodies, against the polypeptides. The antibodies, in turn, are useful for detecting the presence of ULBP polypeptides in human cell samples, which can be correlated with the existence of a malignant condition in a patient. ULBP polypeptides stimulate IFN-γ production, NK cell proliferation, and CTL activity.

The present invention relates to a method for efficiently proliferating and differentiating natural killer cells (NK cells) from umbilical cord blood, more precisely a method for efficiently proliferating and differentiating natural killer cells from umbilical cord blood comprising the following steps: 1) preparing CD3 negative cells by eliminating CD3 positive T-cells from umbilical cord blood derived mononuclear cells; and 2) culturing the CD3 negative cells after treating various cytokines thereto. The present invention is advantageous in obtaining high purity NK cells in a short period of time by inducing NK cells from CD3 negative cells, compared with the conventional method inducing NK cells from haematopoietic stem cells, and in promoting NK cell proliferation and differentiation by treating different cytokines together, for example by the co-treatment of IL-15 and IL-21. That is, the method of the present invention can induce NK cells with increased anti-cancer cytotoxicity, so that it can be effectively used for anti-cancer cell therapy.

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.

Compositions and methods useful in modulating the innate and adaptive immune systems in a subject, including activation of natural killer (NK) cells and/or CD8+ cytotoxic T lymphocytes. The method typically comprises: administering to the subject a composition comprising a therapeutic peptide or a multivalent structured polypeptide comprising multiple copies of the therapeutic peptide described herein in an amount sufficient to increase activity of NK cells and/or CD8+ cytotoxic T lymphocytes in the subject. Preferred therapeutic compositions comprise a carrier; at least one agent selected from the group consisting of: an anti-inflammatory agent, a cytotoxic T cell proliferation agent, or a NK cell proliferation agent; and a therapeutic peptide or a multivalent structured polypeptides of the invention. In certain embodiments, the composition further comprises an immunoglobulin admixed therewith in an amount sufficient to enhance passive immunoprotection in the subject. In other embodiments, the compositions are administered in a therapeutically effective amount to a subject in need thereof to treat rheumatoid arthritis.

The invention relates to the technical field of cell culture, and discloses a high-efficiency NK (natural killer) cell proliferation method. The method is characterized by comprising the following steps: (1) coating a cell culture bottle used for NK cell culture with coating liquid; (2) separating a peripheral blood mononuclear cell (PBMC) containing an NK cell from peripheral blood; (3) adding acell stimulating factor into a cell activation culture solution for PBMC activation; (4) inoculating the activated PBMC into the cell culture bottle after coating treatment in the step (1); and (5) performing induction culture on the PBMC by using an induction culture solution to obtain the NK cell with high activity and purity. The NK cell proliferation method provided by the invention has the characteristics of being simple to operate, high in proliferation efficiency, high in cell activity and high in cell purity; no feeder cells and heterologous serum are added during the culture, therebynot causing medical ethic and GVHD (graft-versus-host disease) risk problems; and the method is applicable to large-scale NK cell preparation and clinical application.

The present invention provides an in-vitro method for inducing proliferation of NK cells, the method comprising contacting a population of NK cells with a nanomatrix, the nanomatrix comprising a) a matrix of mobile polymer chains; and b) attached to said matrix of mobile polymer chains one or more stimulatory agents which provide activation signals to the NK cells; thereby activating and inducing the NK cells to proliferate; wherein the nanomatrix is 1 to 500 nm in size. At least one first and one second stimulatory agents are attached to the same or to separate flexible matrices. If the stimulatory agents are attached to separate beads, fine-tuning of nanomatrices for the proliferation of the NK cells is possible. Closed cell culture systems also benefit from this method.

The invention relates to a culture method for in-vitro induction of an NK cell after recovery of a cryopreserved human PBMC. The culture method comprises three steps including cell recovery, inductionof the NK cell and NK cell proliferation, wherein in the stage of indication of the NK cell, CD 16 and thymosin are diluted with a PBS solution for culture flask enveloping; IL-2, IL-12 and IL-15 areadded to a serum-free culture solution for preparation of an induction medium; IL-2 and IL-7 are added to the serum-free culture solution for preparation of an activation culture medium; and IL-2 isadded to the serum-free culture solution for preparation of a proliferation culture medium, and half-liquid replacement is carried out on the day 3 for solution replacement and activation culture. Themethod for in-vitro induction of the NK cell after recovery of the cryopreserved human PBMC is simple and efficient, reduces the cost of cell culture and also increases safety of cell culture, can effectively overcome the difficulty in inducing the NK cell with the cryopreserved PBMC and deficiency in a proliferation amount, reaches a purpose of cell treatment and enhances resistance to tumors, viruses and infection thereof.

The invention discloses an application of sedge quinone and analogues thereof in culturing NK cells. The invention finds that sedge quinone and analogues thereof, including demethylcyperapuinone and hydroxycyperone, have various pharmacological activities, such as, capability of serving as cryoprotective agents for CIK cells and NK cells, capability of promoting the in vitro multiplication of CIKcells and NK cells and capability of promoting the adipose-derived stem cell osteoblast differentiation. The invention also provides single-open loop derivatives and double-open loop derivatives of the sedge quinone and analogues thereof and a preparation method thereof. The single-open loop derivatives and double-open loop derivatives of the sedge quinone and analogues thereof can promote the invitro multiplication of CIK cells. A searching result shows that the applications of the sedge quinone and analogues thereof in promoting the multiplication of CIK cells and NK cells and promoting thestem cell osteoblast differentiation are not disclosed in the prior art and the single furan ring and double furan ring open loop derivatives of the sedge quinone and analogues thereof are also not reported.

The invention provides an artificial antigen presenting cell applied to efficiently amplifying NK and a construction method thereof. Specifically, the invention relates to fusion protein. The fusion protein is prepared from IL-15, IL-21, MICA protein and optional label sequence and/or signal peptide sequence. Experimental results show that cells of the fusion protein disclosed by the invention canvery well induce NK cells to proliferate, differentiate and activate, and the prepared NK cells has the advantages of large quantity, high purity and strong killing activity. According the artificialantigen presenting cell disclosed by the invention, genes are expressed in serial, so that 1 to 1 100% expression is achieved; meanwhile, operation procedures are simplified; furthermore, time and labor cost are greatly reduced. According the artificial antigen presenting cell disclosed by the invention, the construction method for the NK cell artificial antigen presenting cell is optimized, anda foundation is established for applying the NK cell in cancer therapy, cancer prevention and the like in later.

The invention relates to a preparation method of a portable feeding layer for stimulating NK cell proliferation and differentiation. The preparation method comprises the following steps: (1) collecting 50mL of peripheral blood; (2) separating mononuclear cells by adopting a ficoll density gradient centrifuging method; (3) separating lymphatic and mononuclear cells by utilizing an adherence methodand adhering for 1h; (4) adding factors into lymphocytes and culturing NK cells; (5) carrying out starving culture; after commonly culturing for 10h, collecting the cells. According to the preparationmethod of the portable feeding layer for stimulating the NK cell proliferation and differentiation, provided by the invention, in the aspect of safety, the feeding layer is a macrophage layer inducedby autologous and mononuclear cells and any allosome and heterogeneous cell is not added, so that symptoms including fever caused by extra rejection reaction and the like are not generated.

The invention relates to a method for in-vitro large-scale induction of NK cell expansion, and belongs to the technical field of biology. According to the invention, the constructed novel artificial antigen presenting cell CD86 CD64 mIL-15-aAPC is taken as an expanded feeder cell, NK cells are directly expanded from peripheral blood lymphocytes, and the NK cells expanded by the method has the advantages of good proliferation, high purity, strong cytotoxicity and obvious tumor cell killing effect. The invention solves the problems of limited number of cell expansion, low expression level of surface antigen CD3-/CD56+ and low cell killing activity faced by traditional culture methods, and can meet the clinical requirement on NK cells.

The invention relates to the technical field of cell culture, and discloses a high-efficiency proliferation reagent for peripheral blood NK (Natural killer) cells in vitro and an operation instruction. The reagent comprises the following components: an NK cell proliferation reagent: a gamma-ray inactivated recombinant K562 feeder cell, a CD52 antibody, an IL-2, a CD16 antibody, a CD3 antibody anda CD56 antibody; the gamma-ray inactivated recombinant K562 feeder cell is a K562 cell line for membrane expression of 41BBL and IL-15 which is lethally radiated by a 100G gamma-ray. According to thehigh-efficiency proliferation reagent for the peripheral blood NK cells in vitro and the operation instruction, the prepared NK cells have high purity and killing activity. Through detection, when theNK cells are cultured for 14 days, the purity of the NK cells can reach more than 90%; when the target ratio is 10:1, the killing activity of the NK cells on K562 cells can reach 80% or more; the proliferation speed of the prepared NK cells is high; when the NK cells are cultured for 14-17 days, the cell proliferation rate is more than 500; the NK cells prepared by the invention have excellent effects for targeted killing of tumor cells.

The invention discloses a preparation method and application of a trophoblast modified by genetic engineering, and aims to research whether K562 cells can stably express IL-21, OX40L and CD48 molecules at the same time and enhance NK cell proliferation and tumor killing ability or not. K562 gene engineering cells simultaneously expressed by the three molecules are used as trophoblasts to culture NK, so that the amplification multiple, purity and killing function of the NK are improved; according to the preparation method, K562 cells can be used for simultaneously and stably expressing membrane binding interleukin-21 (mbIL-21), OX40L molecules and CD48 molecules to prepare an engineering cell strain named as K562-ZHX trophoblast cells, then the engineering cell strain is irradiated with cobalt-60 to obtain trophoblast amplified NK cells, the activity of the NK cells prepared by the method is improved by 3 times, the cell amplification can be improved by more than 300 times, a large amount of time and cost can be saved, and a practical foundation can be laid for obtaining high-purity, high-amplification-multiple and high-killing-activity NK cells.

The invention discloses double-open-loop derivatives and preparation methods of cyperapuinone and analogues and application in CIK cell culture. It is found in the invention that the cyperapuinone andits analogues demethylcyperapuinone and hydroxycyperapuinone have multiple types of pharmacological activity, for example, the cyperapuinone and its analogues can serve as cryoprotective agents for CIK and NK cells and can promote in-vitro proliferation of the CIK and NK cells and promote osteogenesis and differentiation of adipose-derived stem cells. The invention also provides single-open-loopderivatives and double-open-loop derivatives and preparation methods of the cyperapuinone and its analogues. The single-open-loop derivatives and double-open-loop derivatives of the cyperapuinone andits analogues can promote in-vitro proliferation of the CIK cells. It is found through retrieval that the application of the cyperapuinone and its analogues in promotion of CIK and NK cell proliferation and osteogenesis and differentiation of adipose-derived stem cells is not disclosed in the prior art and single-furan-ring derivatives and double-furan-ring open-ring derivatives of the cyperapuinone and its analogues is also not reported.

The invention provides a bispecific antibody and an application thereof. The bispecific antibody comprises an IL-21 molecule, an IL-21 molecule and a bispecific antibody, the antibody comprises a CD16a single-chain antibody and an FC, the N tail end of the FC is connected with the C tail end of the single-chain antibody, and the CD16a single-chain antibody comprises a heavy chain variable region and a light chain variable region; and the C terminal of the FC is connected with the N terminal of the IL21 molecule. The bispecific antibody prepared by the invention can target CD16 and IL-21 receptors at the same time and activate NK cells, has a long half-life period, shows stronger anti-tumor and anti-virus capacities compared with a single-target antibody, and has application potentials of promoting NK cell proliferation and being used for in-vitro culture and amplification of NK cells, and the amplified NK cells have good cytotoxicity.

The invention relates to the technical field of medicine, particularly to a lentinan capsule capable of efficiently amplifying NK cells in vivo, and a preparation method thereof, wherein the mushroompolysaccharide capsule comprises 45-80 parts of mushroom powder, 50-75 parts of Dichotomous Fimbristylis Herb powder, 10-15 parts of soybean powder, 3-5 parts of tricalcium phosphate, 1-3 parts of a stabilizer, 0.1-0.25 part of an antioxidant, 0.02-0.05 part of a preservative, and 6-9 parts of a moisturizer. According to the present invention, by combining the Dichotomous Fimbristylis Herb bioactive substance with effects of in vivo NK cell proliferation and differentiation promoting and immunity improving and the lentinan capable of enhancing the activity of NK cells, the obtained lentinan capsule has remarkable antitumor effect; the soybean power added to the formula can increase the absorption of the body to the bioactive ingredients; and the preparation method is simple, the extractedlentinan and the Dichotomous Fimbristylis Herb bioactive substance are high, almost no impurities and harmful substances exist, and the product has high safety and good treatment in tumor treatment.

Compositions and methods for modulating the innate and adaptive immune systems in a subject. The compositions and methods may inhibit the function of inhibitory receptors and enhance activity of activating receptors. The method typically includes the steps of: administering to the subject a composition having a therapeutic peptide or multivalent polypeptide having multiple copies of the therapeutic peptide in an amount sufficient to increase activity of the immune system in the subject. Compositions may include a carrier; at least one agent selected from the group consisting of: an anti-inflammatory agent, a cytotoxic T cell proliferation agent, and/or a NK cell proliferation agent; and a selected therapeutic peptide or multivalent polypeptide. The compositions may also include an immunoglobulin admixed therewith to enhance passive immunoprotection.

The present invention relates to a method of increasing cell proliferation in a population of pancreatic beta cells and a method of treating a subject for a condition associated with an insufficient level of insulin secretion. Also disclosed is a composition. The composition includes a dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1 A) inhibitor and a transforming growth factor beta (TGFP) superfamily signaling pathway inhibitor.

The invention relates to the field of biological medicine, in particular to a bispecific NK cell agonist as well as a preparation method and application thereof. The invention provides a fusion protein, which takes a CD16A receptor and a 4-1BB receptor as targets to obtain a novel bispecific NK cell agonist drug, namely scfv-CD16A-Trimer-4-1BBL, which has the activity of activating and amplifying NK cells in vitro, enhances the antiviral and anti-tumor capabilities of the NK cells, has potential effects of promoting NK cell proliferation and enhancing T cell functions, and can be used for preparing a novel bispecific NK cell agonist drug. The application potential of in-vitro culture and amplification of the NK cells is achieved, and the amplified NK cells have good cytotoxicity.

The invention relates to NK cells with enhanced killing activity and a preparation method thereof. The method includes the steps that a, feeder cells are prepared from peripheral blood mononuclear cells PBMCs; b, NK cell proliferation is conducted, wherein in the step a, the PBMCs are primarily stimulated with monoclonal antibodies CD3 and continue to be activated in a first culture medium containing IL-2, IFN-r, IL-15 and IL-18, primarily activated PBMCs with antigen presentation are obtained and irradiated with gamma rays to be used as the feeder cells, and part of the feeder cells are subjected to freeze preservation with liquid nitrogen; in the step b, CD3+T lymphocytes are removed from the original PBMCs not activated through an immunomagnetic bead sorting method, NK cells after CD56+are gathered again selectively, the NK cells and the feeder cells not subjected to freeze preservation with liquid nitrogen are subjected mixed hatching, NK cell proliferation culturing is conductedin a second culture medium containing IL-2, IL-15, OK432 and nicotinamide, then the NK cells are transferred into a third culture medium containing IL-2 and nicotinamide to be subjected to proliferation culturing, and finally the feeder cells which are thawed from liquid nitrogen freeze preservation are added to continue proliferation culturing.

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to application of six-ingredient pomegranate tablets in preparation of a drug for inhibiting cell proliferation of skin basal cell carcinoma cells A431 and a preparation method of the six-ingredient pomegranate tablets. The six-ingredient pomegranate tablets are prepared from 90 g of pomegranate seeds, 45 g of cortex cinnamomi, 45 g of nutmeg, 45 g of peppers, 45 g of flos carthami and 45 g of semen caesalpiniae cristae. The six-ingredient pomegranate tablets are prepared through supercritical extraction, and the content of piperine is greatly increased.