55 results about "Interleukin 21" patented technology

Methods and compositions for inhibiting interleukin-21 (IL-21) / IL-21 receptor (MU-1) activity using antagonists of IL-21 or IL-21 receptor (“IL-21R” or “MU-1”), are disclosed. IL-21 / IL-21R antagonists can be used to induce immune suppression in vivo, e.g., for treating, ameliorating or preventing autoimmune or inflammatory disorders, including, e.g., inflammatory bowel disease (IBD), rheumatoid arthritis (RA), transplant / graft rejection, psoriasis, asthma, fibrosis, and systemic lupus erythematosus (SLE).

The present invention relates to a composition for expanding lymphocytes comprising at least two types of cytokines selected from interleukin 2 (IL-2), interleukin 15 (IL-15) and interleukin 21 (IL-21). It further relates to a Method of preparing a population of clinically relevant lymphocytes, comprising the steps of: obtaining a body sample from a mammal in particular a tissue sample or body liquid sample, comprising at least one lymphocyte and optionally separating the cells in the body sample, culturing the body sample in-vitro to expand and / or stimulate lymphocytes in the sample wherein the culturing comprises using IL-2, IL-15 and / or IL-21, and optionally determining the presence of clinically relevant lymphocyte in the cultured sample. The present invention also relates to an immunotherapy and the population of clinically relevant lymphocytes.

Methods and compositions for modulating interleukin-21 (IL-21) / IL-21 receptor (MU-1) activity using agonists or antagonists of IL-21 or IL-21 receptor (“IL-21R” or “MU-1”), are disclosed. IL-21 / IL-21R antagonists can be used to induce immune suppression in vivo, e.g., for treating or preventing immune cell-associated pathologies (e.g., pathologies associated with aberrant activity of one or more of mature T cells (mature CD8+, mature CD4+ T cells), mature NK cells, B cells, macrophages and megakaryocytes, including transplant rejection and autoimmune disorders). IL-21 / IL-21R agonists can be used by themselves or in combination with an antigen, e.g., as an adjuvant (e.g., a vaccine adjuvant), to up-regulate an immune response in vivo, e.g., for example, for use in treating cancer and infectious disorders.

Isolated polynucleotides encoding mature interleukin-21, interleukin-21 analogs, polypeptides obtainable from the polynucleotides and uses are disclosed.

The invention belongs to the field of immunology and particularly provides a method for amplifying and activating a natural killer cell (NK) in an oriented way under the mutual effect of a K562 cell, which is transfected by transmembranes IL-21, CD14, CD19, CD86 and CD137, and a low-concentration interleukin 2. According to the invention, the method comprises the following steps: transcribing expression carriers, which are used for stably expressing CD8 alpha-interleukin 21, CD14, CD19, CD86 and CD137, by using K562 cell lines, wherein the CD8 alpha is a membrane protein expressed on a cell membrane, and the interleukin 21 is expressed on the cell membrane to become a transmembrane protein after the CD8 alpha gene is connected with the interleukin 21 gene; and then culturing the K562 cell for a section of time; and finally, obtaining a purified K562 cell by using a physical and chemical method and then culturing the NK cell. According to the invention, by using the amplified and activated NK cell, the immunity of the patient can be enhanced, the viruses and bacteria are resisted, and high efficiency is obtained. The method for amplifying and activating the natural killer cell, provided by the invention, medically has wide purposes.

The invention provides an in vitro amplification method for efficient and highly cytotoxic natural killer (NK) cells. Novel artificial antigen presenting cells 4-1BBL-mIL-21-aAPC, such as 4-1BBL-mIL-21-K562 cells and the like, are constructed through stably expressing 4-1BB ligands (4-1BBL) and membrane immobilized interleukin 21 (mIL-21) on the surfaces of cell membranes, and by using the novel artificial antigen presenting cells as feeder cells for amplification, the NK cells are directly amplified from peripheral blood lymphocytes. Flow cytometry, cytotoxicity test and the like suggest that the amplified cells NK have high purity and strong cytotoxicity and have obvious killing effect on tumor cells.

The invention discloses a gene engineering cell and application thereof in NK (Nature Killer) cell proliferation, and provides an in vitro proliferation method of an NK cell with high efficiency and high cytotoxicity. A 4-1BB ligand (4-1BBL) and a membrane fixed interleukin 21 (mbIl-21) are stably expressed on the surface of a cell membrane by adopting a genetic engineering technology for constructing 4-1BBL-mbIl-21-gene engineering cells (4-1BBL-mbIl-21-Gene Engineering Cells, 4-1BBL-mbIl-21-GEC), such as 4-1BBL-mbIl-21-K562 cells, to be used as feeder cells for proliferation, and the NK cells are directly proliferated from peripheral blood monouclear cells, thus the operation is simpler and easier. Studies such as cell counting, flow cytometry and cytotoxicity tests indicate that high cell proliferation times, high NK cell purity, strong cytotoxicity and remarkable cancer cell killer effect are achieved.

The γc-family cytokines, Interleukin-2 (IL-2), Interleukin-4 (IL-4), Interleukin-7 (IL-7), Interleukin-9 (IL-9), Interleukin-15 (IL-15), and Interleukin-21 (IL-21), are associated with important human diseases, such as leukemia, autoimmune diseases, collagen diseases, diabetes mellitus, skin diseases, degenerative neuronal diseases and graft-versus-host disease (GvHD). Thus, inhibitors of γc-cytokine activity are valuable therapeutic and cosmetic agents as well as research tools. The present embodiments relate to the design of peptide antagonists based on the consensus γc-subunit binding site to inhibit γc-cytokine activity. In several embodiments, peptide antagonists exhibit Simul-Block activity, inhibiting the activity of multiple γc-cytokine family members.

The present invention provides a method for in-vitro culturing and expanding natural killer (NK) cells in a cell culture medium comprising a population of NK cells, the method comprising a) adding an effective concentration of interleukin-21 (IL-21) at the beginning of the culturing process to said medium, b) adding repeatedly an effective concentration of interleukin-2 (IL-2) and / or interleukin-15 (IL-15) to said medium, and c) adding repeatedly feeder cells or membrane particles thereof to said medium, wherein said feeder cells are B cell derived which are EBV immortalized; and wherein said expansion of NK cells in said cell culture medium is maintained for at least 3 weeks.

The invention provides a method for producing recombinant human interleukin-21 by using Pichia pastoris, in particular to a production method of the recombinant human interleukin-21 expressed by the Pichia pastoris, which comprises the following steps: firstly, the reverse transcription PCR (polymerase chain reaction) is carried out in lymphocytes of healthy people to obtain encoding genes of rhIL-21, and the encoding genes are fused in expressional regulatory elements of the Pichia pastoris to construct Pichia pastoris high-level-expression engineering bacteria; and secondly, the Pichia pastoris high-level-expression engineering bacteria are induced to produce a large number of recombinant human interleukins-21 by adding methanol. The Pichia pastoris is very easy to achieve high-density fermentation and has the characteristics of hypersecretion, so that a large number of recombinant human interleukins-21 can be industrially produced easily at low cost.

The present invention provides methods for use of IL-21 in combination with a tyrosine kinase inhibitor (TKI) in treatment of diseases in which inhibition of phosphorylation via TK inhibition and modulation of immune function play a clinically beneficial role. These diseases include, but are not limited to, cancers, such as renal cell carcinoma and metastatic melanoma.

The invention provides a culture medium for in-vitro-amplified-gene edited and activated T cells. The culture medium is characterized by containing a T-cell basal culture medium and further containing: 200IU / ml to 1,000IU / ml of interleukin 7 (IL-7), 300IU / ml to 1,000IU / ml of interleukin 21 (IL-21), 80ng / ml to 120ng / ml of anti-CD40 monoclonal antibody, 0.2mmol / ml to 1.0mmol / ml of serine and 0.3mmol / ml to 1.0mmol / ml of S-2-hydroxyglutarate. The invention further provides a kit containing the culture medium, a method for carrying out in-vitro-amplified-gene editing and activating on T cells by using the culture medium and application in culture of the T cells. By using the culture medium provided by the invention, the gene edited and activated T cells can be extensively amplified under the condition of not adding foreign serum and other T cell activating agents. According to the method provided by the invention, the amplified T cells can be activated by 500 to 1,000 times, and the times of activation are apparently higher than that in the prior art.

The present invention relates to an agent for differentiating hematopoietic stem cells into natural killer cells and a method for the differentiation, more precisely an agent for differentiating hematopoietic stem cells into natural killer cells comprising YC-I [3- ( 5'-hydroxymethyl-2'-furyl)-1-benzylindazole] or IL-21 (Interleukin-21) as an active ingredient and a method for differentiating hematopoietic stem cells into natural killer cells using the same. The YC-I and IL-21 regulate the differentiation of hematopoietic stem cells into natural killer cells and increase the killing activity of NK cells. Therefore, the agent for NK cell differentiation of the present invention can be effectively used for cell therapy for the treatment of cancer by regulating the differentiation of hematopoietic stem cells into natural killer cells having tumor cell killing activity.

A method of carrying out adoptive immunotherapy by administering a subject an antigen-specific cytotoxic T lymphocytes (CTL) preparation in a treatment-effective amount is described. In the method, the CTL preparation is preferably administered as a preparation of an in vitro antigen-stimulated and expanded primate CTL population, the CTL population: (i) depleted of FoxP3+ T lymphocytes prior to antigen stimulation; (ii) antigen-stimulated in vitro in the presence of interleukin-21; or (iii) both depleted of FoxP3+ T lymphocytes prior to antigen stimulation and then antigen-stimulated in vitro in the presence of interleukin-21. Methods of preparing such compositions, and compositions useful for carrying out the adoptive immunotherapy, are also described.

A conjugate protein comprising a GM-CSF or fragment thereof linked to an IL-21 or fragment thereof is described. The conjugate protein has unexpected immune activating and tumoricidal properties and is useful in a variety of therapeutic applications.

The invention discloses an in-vitro culture method of a central memory T lymphocyte. The method comprises the following steps: adding IL-21 into an initial medium, and allowing the oxygen concentration in a culture environment to be 1 + / - 0.1%; and adding the IL-21 and rapamycin into an amplification medium when the culturing is performed for 7 days. The in-vitro culture method using the combination of the rapamycin, interleukin 21 and low-oxygen environment in a proper culture process to improve the proportion of the central memory T cell, inhibit the differentiation of the central memory T cell into effector cells and enhance the killing effect on tumor cells is disclosed for the first time.

The invention provides an IL7 (Interleukin-7) and IL21 (Interleukin-21) modified NK92 (Natural Killer-92) cell as well as a preparation method and application thereof. Particularly, the NK cell provided by the invention can be used for effectively killing a tumor cell, is obviously enhanced in in-vitro multiplication capacity, is easily proliferated in vitro, can be cultured in a high-density manner, has quite high killing capacity, a low production code and a few side effects, and does not have a risk of a graft versus host disease. Moreover, the NK cell can be used for obviously stimulatingthe activation and the amplification of a T (thymus) cell to cooperate with each other for jointly killing the tumor cell; the immune response of an organism is effectively enhanced; meanwhile, no cytotoxicity exists between the NK cell and the T cell each other, and a respective killing effect on a target cell is also not influenced.

The invention relates to the technical field of tumor treatment, specifically to recombinant mesenchymal stem cell and application thereof. The recombinant mesenchymal stem cell belongs to mesenchymalstem cell used for killing tumor formed by integrating cDNA for expressing interleukin-21 to gene of mesenchymal stem cell, which can perform effective positioning the position of a tumor, can perform high-level expression of interleukin-21, can rapidly activate immune cell through secreted interleukin-2 after accurately positioning the position of the tumor, rapidly and effectively inhibit and kill tumor cell, and has very important research significance in the treatment field of tumor and cancer.

The invention provides a kit for culturing natural killer cells in vitro and a use method and application of the kit. The kit comprises a coating solution, an incubation solution, a serum-free culturemedium 1, a serum-free culture medium 2, a serum-free culture medium 3 and a serum-free culture medium 4, wherein the serum-free culture medium 1, the serum-free culture medium 2 and the serum-free culture medium 3 are serum-free culture media containing interleukin 2, interleukin 15, interleukin 18, interleukin 21 and autologous plasma, and the serum-free culture medium 4 is a serum-free culturemedium comprising interleukin 2, interleukin 15, interleukin 18 and interleukin 21. By reasonably matching the types and concentrations of autologous plasma and interleukin in each culture medium, high-yield and high-purity NK cells can be efficiently prepared under the conditions of fewer seed cells and no separation and purification.

The invention discloses an immune cell culture medium, a culture method and application. The immune cell culture medium comprises a base cell culture medium, cell factors and glucocorticoid; the immune cell culture method comprises the following steps: S1, collecting and activating immune cells to be cultured; S2, inoculating the activated immune cells into the immune cell culture medium for culturing the cells; S3, harvesting the immune cells. A cell culture stimulating liquid comprises the base cell culture medium and the cell factors; the cell factors comprise interleukin 2, interleukin 15and / or interleukin 21. According to the immune cell culture medium and the culture method provided by the invention, the proportion of CD8+NK cells in the cultured NK cells can be effectively increased.

The present invention discloses applications of interleukin-21 in preparation of a stem cell-like memory T cell in vitro amplification inducer. According to the present invention, with the addition of interleukin-21 as an inducer to a T cell induction culture medium, the differentiation amplification of Naive CD8+ T cells can be induced to obtain TSCM cells, the forming proportion and the absolute number of the TSCM cells are significantly increased, and the effect is significantly superior to the effect of the inducer IL-2 universally used in the cell adoptive immunotherapy; and the interleukin-21 is the natural cytokine secreted by human body, has the high safety, does not have the drug toxicity and other problems, can provide the good technical support for the extensive development of the adoptive immunotherapy, and has the important promotion significance.

The invention belongs to the fields of genetic engineering and biomedicines and relates to a novel medicinal application of interleukin 21 (IL-21), and particularly relates to an application of the IL-21 in preparation of an anti-HBV preparation. Tests prove that by using the IL-21 for intervening in a HBV chronically infected mouse model, conversion rate of hepatitis B surface antigen (HBsAg) andhepatitis B e antigen (HBeAg) can be significantly increased, and meanwhile, DNA carrying quantity of HBV in serum is significantly reduced. The IL-21 can be used as a medicine precursor for developing a novel anti-hepatitis B virus medicine.

The invention provides a method of treating or preventing viral diseases in a mammal comprising administering to the mammal an interleukin (IL)-21 blocking agent in an amount effective to treat or prevent the viral disease in the mammal. Also provided is a method of reducing the activation or recruitment of immune cells in a mammal comprising administering to the mammal an IL-21 blocking agent in an amount effective to reduce the activation or recruitment of immune cells in the mammal. Methods of decreasing the expression of at least one cytokine or at least one protein in a mammal comprising administering to the mammal an IL-21 blocking agent in an amount effective to decrease the expression of the cytokine or the protein are also provided.

The invention discloses a human interleukin 21 nucleotide sequence and the method to produce the mature interleukin 21 and the application. The nucleotide sequence is SEQ ID No.1. The manufacture includes the following steps: making SEQ ID No.1, inducing into expression carrier to form reconstruction carrier, transforming the carrier into prokaryotic cell to cultivate and multiple, taking inducing, distilling, purification, the mature human interleukin 21 of SEQ ID No.2 would be gained. The invention could directly express human interleukin 21 in prokaryotic cell. It could be used to produce gene engineer medicine.

The invention relates to the field of cancer therapy and relates to fusion protein of interleukin 21 and an anti-Her2 / neu single-chain antibody, wherein the fusion protein is utilized to treat breast cancer.

The invention discloses a method for inducing initial B cells to differentiate into regulatory B cells in an in-vitro way and culture conditions thereof. The method comprises the following step of performing induced culture on the initial B cells in an inducing culture medium, wherein the inducing culture medium consists of an RPMI-1640 culture medium, fetal calf serum, lipopolysaccharide, a CD40ligand, interleukin 1 beta, interleukin 6, interleukin 4, interleukin 21 and interleukin 23. The method has the advantages that the initial B cells are cultured by the inducing culture medium, so thatthe enough number of regulatory B cells (CD19<+>CD24<hi>CD27<+>) is obtained; the ability of inducing the initial B cells to differentiate into the regulatory B cells (CD19<+>CD24<hi>CD27<+>) is obviously improved, the survival rate of the regulatory B cells (CD19<+>CD24<hi>CD27<+>) after inducing differentiating is increased, and the good application prospect is obtained.