183 results about "CD16" 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.

Methods for the identification and isolation of multipotent cells from non-osteochondral mesenchymal tissue. Specifically, this invention relates to an adult multipotent cell or a cell population or composition comprising said cell, isolated from non-osteochondral mesenchymal tissue, characterized in that the cell is positive for the following markers: CD9, CD10, CD13, CD29, CD44, CD49A, CD51, CD54, CD55, CD58, CD59, CD90 and CD105 and because it lacks expression of the following markers: CD11b, CD14, CD15, CD16, CD31, CD34, CD45, CD49f, CD102, CD104, CD106 and CD133.

A method is provided for enhancing ADCC in an individual in need thereof, comprising the administration of T lymphocytes expressing a CD16-like receptor in said individual. Said method for enhancing ADCC may be used to treating cancers, autoimmune diseases or infections.

Identification and isolation of multipotent cells from non-osteochondral mesenchymal tissue. This invention relates to the identification and isolation of multipotent cells from non-osteochondral mesenchymal tissue. Specifically, it relates to an adult multipotent cell or a cell population or composition comprising said cell, isolated from non-osteochondral mesenchymal tissue, characterized in that it is positive for the following markers: CD9, CD10, CD13, CD29, CD44, CD49A, CD51, CD54, CD55, CD58, CD59, CD90 and CD105 and because it lacks expression of the following markers: CD11b, CD14, CD15, CD16, CD31, CD34, CD45, CD49f, CD102, CD104, CD106 and CD133.

The invention concerns the use of human or humanized chimeric monoclonal antibodies which are produced in selected cell lines, said antibodies bringing about a high ADCC activity as well as a high secretion of cytokines and interleukins, for treating underpopulations of so-called weak-response patients exhibiting CD16 FCGR3A-158F homozygote or FCGR3A-158V / F heterozygote polymorphism.

The invention discloses a method for simultaneous and efficient amplification of CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells. The method comprises the steps as follows: the concentration of separated PBMC (peripheral blood mononuclear cells) is adjusted by a serum-free medium containing autologous plasma, an Anti-CD16 antibody, IL-2 and IL-15 are added, and then the mixture is transferred into a T175 culture flask for culture; an Anti-CD3 antibody and an Anti-CD137 antibody are added; a serum-free medium containing the autologous plasma, IL-2 and IL-15 is supplemented every two days according to the cell growth condition; the cell concentration is controlled to be about 1.5*10<6> / ml; and after culture is performed for 14-21 days, large quantities of high-purity CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells can be obtained simultaneously, and the total cell quantity can reach an effective value of the cell quantity required for adoptive cellular immunotherapy clinically for tumor. The method for simultaneous and efficient amplification of the CD<3+>CD<56+>CIK cells and the CD<3->CD<56+>NK cells is simple, convenient, effective and high in cell killing activity.

The invention relates to the field of stem cell biology, specifically to a method for differentiating human pluripotent stem cells into natural killer cells and an application. The invention disclosesa pluripotent stem cell-derived natural killer cell. The pluripotent stem cell-derived natural killer cell expresses CD56, Nkp30, Nkp44 and Nkp46, and also expresses markers CD16 and CD94 of a maturenatural killer cell. The invention also discloses a method for preparing the natural killer cell. The method comprises the following steps: S1, formation of an embryoid body; S2, differentiation of the embryoid body into hematopoietic progenitor cells; S3, differentiation of the hematopoietic progenitor cells into NK cells; and S4, maturation and expansion of the NK cells. With the differentiation method provided by the invention, the pluripotent stem cells can be rapidly, efficiently, simply and conveniently induced to be differentiated into the natural killer cells with low cost on the basis of a culture medium with definite components and an optimized cell factor combination.

A human monocyte cell characterised by the expression of the following markers: Tie2, CD16 and CD14.

The invention relates to a method for abundantly amplifying NK (Natural Killer) cells from a mononuclear cell of peripheral blood. The method comprises the following steps of collecting the mononuclear cell of the peripheral blood, first putting the mononuclear cell into a culture bottle enveloped by a CD16 monoclonal antibody, culturing the mononuclear cell, and adding IL-2, IL-15, OK432 and inactivated autoserum into a culture medium; subsequently, transferring the cell into a culture bottle which is not enveloped by the CD16 monoclonal antibody, culturing the cell, adding the IL-2, the IL-15 and the inactivated autoserum into the culture medium; afterwards, transferring the cell into a culture bag, culturing the cell, and adding IL-4 into the culture medium on the last third day, and continuously culturing the cell, so as to obtain abundant NK cells. According to the method, components of animal serum and a tumor cell are not contained; therefore, the safety and the reliability of the NK cells are improved to a great extent; the method has a favorable application prospect.

The invention relates to a method for efficiently amplifying natural killer cells from peripheral blood in a high purity manner. The method comprises the following steps: collecting 100ml-200ml of peripheral blood, separating mononuclear cell, putting the mononuclear cell into a culture bottle coated with CD16 monoclonal antibody and CD226 monoclonal antibody for culturing, and activating the cell by virtue of IFN-gamma; after the activation is carried out for 24 hours, adding IL-2, IL-15, IL-21, OK432, CD2 monoclonal antibody and inactivated autoserum into a culture medium; transferring the cell into a culture bottle which is not coated with CD16 monoclonal antibody and CD226 monoclonal antibody for culturing, and supplementing liquid to the cell once in each 2-3 days; and finally transferring the cell into a culture bag for culturing, and collecting NK cell in the 21th day. According to the method, no any heterologous serum component or trophoblast cell is used, so that the safety and reliability of the prepared NK cell are guaranteed; and the method has good clinical application values.

The invention provides a method of enumerating the number of cells of a cell type in a cell sample by (a) counting the white blood cells in the cell sample to obtain the white blood cell population of the sample; (b) determining the proportion or percentage of the cells of the cell type in the white blood cell population in the sample; and (c) calculating the number of cells of the cell type in the sample. The cell type may be a lymphocyte sub-set selected from the group comprising CD4+ lymphocytes, CD 45 cells, CD19 cells, CD16 and CD56 positive cells, CD8 cells, CD3 cells or any combination thereof. The method is particularly useful in monitoring the immune status of a patient infected with HIV or other immune deficiency state or disease or condition where CD4+ lymphocytes or CD4+ T cells are monitored or counted.

The invention relates to an in-vitro pure culture method for natural killer cells. The method includes the following steps that peripheral blood is collected, single karyocyte is separated, magnetic beads combined with specific antibodies are adopted to remove the NK cells, and NK cell populations are obtained; the NK cell populations are put into a culture bottle coated with multiple antibodies,and are subjected to activation culture with a complete medium containing IL-2, IL-7, IL-15, anti-CD16, OK432 and inactivated autologous serum, and inactivated NK cell populations are obtained; the inactivated NK cell populations are put into a culture bottle free of antibody coating and continue to carry out multiplication culture, and the natural killer cells are obtained. Any heterologous serumingredients and trophoblast cells are not used in the in-vitro pure culture method, and high amplification efficiency can be wholly obtained; meanwhile, the purity of the amplified NK cells reaches 90% or above, in-vitro efficient pure culture is achieved, and the clinical application requirement is conveniently met.

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 invention relates to a method for in vitro multiplication culture of NK cells. The method comprises coating a culture bottle with heparin sodium, CD16 and human immunoglobulin, collecting peripheral blood, inactivating plasma, separating mononuclear cells, using a serum-free medium (containing autologous plasma, IL-2, IL-15 and IL-18) to culture multiplication cells, harvesting and detecting the cells and the like. Trophoblast cells or magnetic beads are not used for sorting and purifying, and the method has simple operation, high safety, good amplification effect and high clinical application value.

Provided is a method of culturing self-activated lymphocytes applicable to the treatment of malignant tumors. The method raises the percentage's of natural killer (NK) cells of the lymphocytes and evenly activate the NK cells, T cells and natural killer T (NKT) cells, and thus can be used to effectively eliminate various kinds of cancer cells. The method of culturing self- activated lymphocytes involves: extracting lymphocytes from human peripheral blood; performing a first culturing step of culturing the extracted lymphocytes in a culture fluid to which IL-2, L-glutamine and autochthonous plasma are added, in the presence of anti-CD3, anti- CD 16, and anti-CD56 antibodies; and performing a second culturing step of culturing the mixed culture fluid resulting from the first culturing step in the presence of anti-CD3, anti-CD16, and anti-CD56 antibodies after being admixed with a culture fluid to which IL-2, L-glutamine and autochthonous plasma are added.

It is intended to provide a method for producing an NK cell-enriched blood product, the method being less invasive and capable of conveniently and rapidly growing NK cells, etc. in blood collected from an organism. The NK cells in blood are stimulated with NK cell growth-stimulating factors comprising an anti-CD16 antibody, OK432, a bisphosphonate derivative or a salt thereof, or a hydrate thereof, and a cytokine. Then, the blood is cultured at a physiological cell temperature to produce an NK cell-enriched blood product.

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 a preparation method of a cytokine-induced killing cell for inducing antibody-dependent cellular cytotoxicity. The preparation method comprises constructing an interferon gamma signal peptide-tumor cell combined peptide, bonding 2th and 3th constant region gene segments of a crystallizable fragment domain heavy chain to the combined peptide, recombining the combined chain to a viral vector, transfecting a human cytokine-induced killing cell and carrying out high expression of a 2th and 3th constant region fusion protein of a novel polypeptide-crystallizable fragment domain heavy chain. The preparation method can cause antibody-dependent cellular cytotoxicity, improve a cytokine-induced killing cell proliferation multiple to 1000 times or more, a CD3+ / CD56+ expression rate to 40% or more and a CD16+ expression rate to 30% or more. In-vivo and in-vitro tests prove that the cytokine-induced killing cell has strong tumor killing toxicity. The invention also provides a kit for autologous cytokine-induced killing cell proliferation culture and has a high-efficiency anti-tumor function.

The invention discloses a fusion protein of Her2 antibody and interleukin 2 and application thereof, and belongs to the technical fields of medical science and oncology phymatology. The fusion protein comprises CD16 monoclonal antibody heavy chain signal peptide, ErbB2 antibody heavy chain variable region, a Linker 1, a light chain variable region, a human antibody Fc segment, a Linker 2 and IL-2 mature peptide, and has the amino acid sequence shown as SEQ ID No.1. The fusion protein can inhibit Herceptin resistant breast cancer cell proliferation, and can kill Her2 high-expression breast cancer cells. The invention lays a foundation for clinical application of the fusion protein of the Her2 antibody and the interleukin 2 in the anti-tumor aspect.

Provided is a medium composition for culturing self- activated lymphocytes applicable to the treatment of malignant tumors, to which anti-CD3, anti-CD16 and anti-CD56 antibodies are added along with interleukin2 (IL-2) to evenly activate natural killer (NK) cells, T cells and natural killer T (NKT) cells, and thus the medium composition can be used to culture im- munocytes that can effectively treat various kinds of malignant tumors. The medium composition includes a cell culture medium and additives added to the cell culture medium, wherein the additives include interleukin2 (IL-2), anti-CD3 antibodies, anti-CD16 antibodies, and anti-CD56 antibodies.

The invention belongs to the technical field of cells, and particularly relates to a cell culture solution and cell culture method and application thereof. The cell culture solution comprises a first culture solution and a second culture solution, the first culture solution contains cell factors such as OKT3, CD16, IL-2, IL-15 and 4-1BBL, and the second culture solution contains cell factors such as IL-2, IL-15 and 4-1BBL. The invention further provides the cell culture method. The method comprises the steps that cells are inoculated into the first culture solution for culture, and then the first culture solution is supplemented on the next day; the second culture solution is supplemented when culture is conducted on the ninth day for continuous culture, and the second culture solution is supplemented on the next day. According to the cell culture solution and cell culture method and application thereof, the cell factor combination and culture technological process is optimized, the culture period is shortened, only 14-21 days are needed for culture, and the purity of NK cells obtained through culture reaches 70% or above; the NK cells do not need to be purified, the steps are simple and convenient, and the operability is high.

Disclosed is a medium composition for culturing self-activated lymphocytes, which contains anti-CD3 antibody and anti-CD16 antibody in addition to interleukin 2 (IL-2), interleukin 12 (IL-12) and interleukin 18 (IL-18) in a medium, and thus can efficiently proliferate and activate NK cells, T cells and NKT cells and, at the same time, can significantly increase the ratio of NK cells in lymphocytes so as to provide immunocytes having excellent effects on the treatment of various kinds of malignant tumors, and a method for culturing self-activated lymphocytes using the medium composition.

The invention provides a method for preparing efficient clinical-level CD 56<+> group immune cell. The method comprises the following steps: a) separating single karyocyte from peripheral blood or umbilical cord blood; b) inoculating the single karyocyte in the step a) to an anti-CD16 antibody-coated culture dish; c) adding rabbit-anti-human thymic cell immune globulin, Lifein and animal origin-free cytokine in the culture dish for inducing and stimulating the single karyocyte inoculated in the step b); d) supplementing the culture dish and the animal origin-free cytokine according to the cells counting results every 2-3 days; and e) obtaining the CD 56<+> group immune cell. The method can reduce the cost, accords with the requirement of clinical-level preparation, and has good killing activity in vitro and vivo.

It is intended to provide a method for producing an NK cell-enriched blood preparation, which is low invasive and is capable of conveniently and rapidly growing NK cells, etc. in blood collected from an organism. The NK cells in blood are stimulated with NK cell growth-stimulating factors comprising an anti-CD16 antibody, OK432, an anti-CD3 antibody, and a cytokine. Then, the blood is cultured at a physiological cell temperature to produce an NK cell-enriched blood preparation.

Disclosed is a medium composition for culturing self-activated lymphocytes, which contains anti-CD3 antibody and anti-CD16 antibody in addition to interleukin 2 (IL-2), interleukin 12 (IL-12) and interleukin 18 (IL-18) in a medium, and thus can efficiently proliferate and activate NK cells, T cells and NKT cells and, at the same time, can significantly increase the ratio of NK cells in lymphocytes so as to provide immunocytes having excellent effects on the treatment of various kinds of malignant tumors, and a method for culturing self-activated lymphocytes using the medium composition.

The invention discloses a detection method of human peripheral blood lymphocytes. The preparation method comprises the following steps of: taking human anticoagulation peripheral blood; adding anti-human CD3, CD4, CD8 and CD45 monoclonal antibodies and anti-human CD3, CD56, CD16 and CD45 monoclonal antibodies, then adding a hemolysin working solution containing a fluorescent probe to respectivelyobtain the percentage and absolute count value of a human peripheral blood T lymphocyte subset and an NK lymphocyte subset and achieve the detection of human peripheral blood lymphocytes. The method has the advantages of simplicity and convenience in operation, long sample stable preservation time, high accuracy, multi-index output and the like.

The invention discloses an efficient in-vitro amplification method of human natural killer (NK) cells. The efficient in-vitro amplification method includes the steps that CD16 monoclonal antibodies, ATG and IL-2 are added into a processed culturing system, incubation is carried out in an incubator at the temperature of 37 DEG C, counting and solution changing are carried out, and fresh IL-2 is added; a culture solution is changed at regular intervals, culturing continues for a proper time, and the human NK cells are obtained. The efficient in-vitro amplification method has the advantages that the NK cells can be efficiently amplified through the ATG and the CD16 antibodies in a short time without being subjected to separation of magnetic beads and irritation of tumor modification cells, production cost is greatly saved, and operation is easy and convenient; the number of amplification times of the NK cells is one thousand or more, and the final proportion ranges from 80% to 93%; production efficiency is high, and the efficient in-vitro amplification method can be entirely applied to large-scale clinical researching and the like; the growth process of the NK cells is completed under the GMP condition, sterile operation is guaranteed, various kinds of security detection are carried out at the final production stage, and the efficient in-vitro amplification method can be applied to clinical researching.

The invention provides a preparation method of autologous natural killer cell proliferation and culture. The preparation method of the autologous natural killer cell proliferation and culture is characterized in that mononuclear cells of a cancer patient are amplified and activated to obtain natural killer cells after the mononuclear cells are transfected by 3- phosphoinositide-dependent protein kinase-1 and CD122 and under the effect of K562 cells which are expressed stably serve as trophoblast cells and interleukin 2, proliferation times of the natural killer cells are above 2500, phenotype of CD16+ / CD56+ is above 90%, and in-vivo and in-vitro tests show that the natural killer cells have an anti-tumor and have quite high killer toxicity. The invention also provides a kit which is used for autologous natural killer cell proliferation and culture and is obtained by using the preparation method. The kit has an efficient anti-tumor function.