188 results about "CD3 Antibody" patented technology

The present invention provides methods of treating, preventing, slowing the progression of, or ameliorating the symptoms of T cell mediated immunological diseases, particularly autoimmune diseases (e.g., autoimmune diabetes (i.e. type 1 diabetes or insulin-dependent diabetes mellitus (IDDM)) and multiple sclerosis) through the use of anti-human CD3 antibodies. The antibodies of the invention of the invention are preferably used in low dose dosing regimens, chronic dosing regimens or regimens that involve redosing after a certain period of time. The methods of the invention provide for administration of antibodies that specifically bind the epsilon subunit within the human CD3 complex. Such antibodies modulate the T cell receptor/alloantigen interaction and, thus, regulate the T cell mediated cytotoxicity associated with autoimmune disorders. Additionally, the methods of the invention provide for use of anti-human CD3 antibodies modified such that they exhibit reduced or eliminated effector function and T cell activation as compared to non-modified anti-human CD3 antibodies.

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 discloses culturing, freezing and recovering methods of an activated lymphocyte with CD3+CD8+as major, which can solve problems that a patient needs carrying out blood sampling for many times caused by continuously utilizing the activated lymphocyte with CD3+CD8+as major. The method comprises the following steps of: (1) contacting the extracted lymphocyte of the peripheral blood with IL-2, IL-15, an anti-CD3 antibody and an anti-CD28 antibody, so as to amplify the activated lymphocyte with CD3+CD8+as major; (2) freezing and storing the activated lymphocyte; and (3) recovering the activated lymphocyte. The activated lymphocyte cultured via the method disclosed by the invention has clear components, and comprises few CD4+CD25+Treg cells and more CD8+T lymphocyte; feedback time and frequency of the activated lymphocyte can be adjusted according to other treatments for a patient, such as a radiotherapy or a chemotherapy, so that diseases, such as tumor, infectious diseases and immunodeficiency can be treated well.

Non-toxic anti-CD3 antibody is useful for the treatment, prevention, and reversal of human autoimmune disease. Anti-CD3 antibody is generated by immunizing xenogenic mice capable of developing fully human antibodies. Because the inventive antibodies are not derived from other species, they do not the invoke the immune responses typically associated with humanized or grafted antibodies.

The invention provides a culture system and a culture method for efficient amplification in vitro of autologous NK (Natural Killer) cells. The culture system comprises a basic culture medium containing GT-T551, interleukin IL-2, interleukin 1L-12, interleukin IL-15, interleukin IL-18, interleukin 1L-17, interleukin IL-22 and natural traditional Chinese medicine substances. The culture method comprises the following steps of: collecting and separating autologous 40ml peripheral blood mononuclear cells; resuspending the cells by a culture medium containing 10 percent of self plasma; adding the cells into a cell culture bottle coated by CD3 antibodies or CD56 antibodiees; performing stimulation by a cell factor combination A; putting the cells in a 37 DEG C and 5-percent-CO<2> culture tank to perform incubation and shaking; after 6 to 7 days, transferring the cells into a cell culture bag to be cultured; complementally adding a culture medium containing a cell factor combination B; in the culture period, culturing the cells in a basic culture medium containing IL-2 every 3 to 4 days; and obtaining high-purity NK cells after the total culture time of 17 to 19 days. The culture system and the culture method have the advantages that the NK cells can be amplified by more than 1000 times in vitro; and the high-purity NK cells exceeding 5*10<9> can be obtained.

The invention provides a new cell sorting magnetic bead, which is a 50-70 nm gold plated Fe3O4 magnetic nanoparticle (Fe3O4 and Au nanoparticle) ostensibly coupled with proteinA by covalent bonds, interacted with the specificity of a Fc section of IgG through the proteinA, and directionally coupled with IgG type mouse CD3 antibodies. The invention relates to a method for synthesizing and modifying the magnetic bead, and a method for carrying out cell sorting by using the magnetic bead. The magnetic bead of the invention is used for removing the CD3 T cells in the mouse splenocytes so as to reduce the CD3 T cell content of the splenocytes from 36.5 percent to 0.7 percent, and has extremely high sorting efficiency. The magnetic bead also has the advantages of high protein-coupled quantity and superparamagnetism, and the preparation method has the advantages of simpleness, environmental protection, low cost and good application prospect.

The invention provides a NK (natural killer) cell culture medium and a culture method of a NK cell, wherein the NK (natural killer) cell culture medium comprises the following components in parts by weight: 83 to 93 parts of serum-free basal culture medium; 4 to 6 parts of plasma; 0.5 to 1.5 parts of interleukin-2; 0.5 to 1.5 parts of anti-CD3 monoclonal antibody; 1 to 5 parts of insulin-like growth factor 1; 1 to 3 parts of lycium barbarum polysaccharide. According to the NK (natural killer) cell culture medium provided by the invention, the safety is better, and the NK cell cultured by the culture medium with the components can quickly and stably proliferate, and has better killing activity to tumor cell. Experimental results show that, after using the NK cell culture medium provided by the invention for culturing the NK cell for two weeks, the proliferation times soars almost fiftyfold; the killing activity to K562 reaches 91 percent at effector-target ratio being 40: 1.

The invention discloses a preparation method of human TSCMs (T memory stem cells), which comprises the following steps: (1) collecting peripheral venous blood with a blood cell separator or injector under aseptic conditions, and carrying out Ficoll-Hypaque density gradient centrifugation to obtain mononuclear cells; (2) putting the separated PBMCs (peripheral blood mononuclear cells) in a culture medium containing irritants and cell factors, regulating the cell density to (0.5-2)*10<6>/mL, transferring into a cell culture plate, culture bottle or culture bag, and culturing in an incubator; (3) after culturing the cells for 48-72 hours, replacing half of the culture solution, supplementing the equivalent culture solution to keep the cell density at (0.5-2)*10<6>/mL; and according to the cell growth state, harvesting the cells in the 10th-21st day. An anti-CD3 monoclonal antibody and an anti-CD28 monoclonal antibody are utilized as the irritants to activate the TSCMs, the cell factors rh IL-7, rh IL-15 and rh IL-2 are combined for irritation, and IM-12 is utilized to stop the stem cell differentiation, so that the cultured TSCMs have the common characteristics of both memory cells and stem cells, thereby providing a new optional scheme for clinical adoptive immunity cellular therapy.

Provided herein are compositions and methods for the treatment of type 1 diabetes (T1D) in mammalian subjects. The compositions include lactic acid fermenting bacteria (LAB) expressing an IL-2 gene and a T1D-specific self-antigen (e.g., proinsulin (PINS)) gene. Exemplary methods include: orally administering to a mammalian subject, a therapeutically effective amount of the composition. The composition can be administered to the subject mucosally, resulting in delivery of the LAB into the gastrointestinal tract, where the LAB is released. Bioactive polypeptides expressed by the LAB are thus administered via mucosal delivery. The LAB may be selected to deliver a low-dose of IL-2 to the subject. The methods may not require concomitant systemic anti-CD3 antibody treatment. The methods may be suited for subjects possessing residual beta-cell function, e.g., those with recent-onset T1D.

The invention provides a kit for assessing a function of cytotoxic immune cells in human peripheral blood and an assessment method. The kit for assessing the function of cytotoxic immune cells in human peripheral blood provided by the invention comprises an anti-human CD3 antibody, an anti-human CD8 antibody, an anti-human CD45RA antibody, an anti-human CCR7 antibody, an anti-human CD28 antibody,an anti-human CD38 antibody, an anti-human CD57 antibody, an anti-human HLA-DR antibody, an anti-human PD-1 antibody, an anti-human CD56 antibody, an anti-human CD94 antibody, an anti-human NKP30 antibody, an anti-human NKP46 antibody, an anti-human NKG2D antibody, an anti-human KIR antibody, an anti-human Gamma Delta antibody and an anti-human V Delta 2 antibody, wherein all the antibodies are labeled by fluorescein. The kit for assessing the function of cytotoxic immune cells in human peripheral blood provided by the invention can be used for comprehensively assessing the immunity function of the cytotoxic immune cells in human peripheral blood, and is convenient and safe for use.

The present invention relates to humanized or chimeric antibodies binding CD3. It furthermore relates to bispecific antibodies, compositions, pharmaceutical compositions, use of said antibodies in the treatment of a disease, and method of treatment.

The invention relates to a method for in-vitro culture of killer T cells. Specifically, IL-2 and an anti-CD3 monoclonal antibody are combined with indigowoad leaf decoction for costimulation of lymphocytes taken from 50ml of in-vitro blood so as to get the T cells with cytotoxin activity by induction; and the T cells with the cytotoxin activity, which are obtained by an induction culture method, have the advantages of large proliferation number and good vitality, and the number of the CD8-positive T cells can be increased from 20%-30% to 80%-90%. Simultaneously, the method disclosed by the invention has the advantages of low taking quantity of the blood, simple operation steps, low cost of a reagent and short maturation time of the cells, above 1*10<9> cells can be achieved by 8 days and used for performing continuous intravenous infusion on a patient six times, and above 1*10<9> cells can be infused every time. The method is suitable for a clinical biological immunotherapy of the patients with tumors and can be widely popularized and applied.

An NK cell culture medium is prepared from, by weight, 94-98 parts of serum-free basic culture medium, 7-10 parts of plasma, 0.6-1.6 parts of anti-CD3 monoclonal antibody, 2-4 parts of first para-insulin growth factor, 20-30 parts of PBS buffer solution, 1-4 parts of squid ink, 2-6 parts of flos carthami polysaccharide, 2.5-4.5 parts of allicin, 1-3 parts of alfalfa polysaccharide and 2-4 parts of cinnamaldehyde. Serum-free basic culture medium, plasma, anti-CD3 monoclonal antibody and first para-insulin growth factor together form a human body marrow environment required by NK cell growth. Flos carthami polysaccharide, allicin, alfalfa polysaccharide and cinnamaldehyde can improve activity of the NK cells easily, and accordingly killing capacity of the NK cells is enhanced. Squid ink contains IL-2 which can induce activation and differentiation of the NK cells, and accordingly multiplication efficiency of the NK cells is easily improved. The activated NK cells can synthesize and secrete various cytokines, and accordingly tumor cell differentiation and multiplication are directly inhibited.

The invention relates to an induced culture composition capable of promoting proliferation of a spleen-derived CD8 positive T cell and application of the induced culture composition. The induced culture composition comprises a culture medium and inducing factors added in the culture medium, the inducing factors comprises phytohemagglutinin, a CD3 antibody and a CD28 antibody, the final concentration of the phytohemagglutinin is 1 mu g/mL to 10 mu g/mL, the final concentration of the CD3 antibody is 1 mu g/mL to 10 mu g/mL, the final concentration of the CD28 antibody is 1 mu g/mL to 10 mu g/mL. The induced culture composition provided by the invention can promote proliferation of the spleen-derived CD8 positive T cell.

The invention discloses a kit for detecting mycobacterium tuberculosis infection by using peripheral blood. The kit provided by the invention comprises the following substances: (1) a mycobacterium tuberculosis Rv3615c mixed polypeptide library which consists of all or a part of 19 polypeptides consisting of amino acid sequences shown as sequences 1 to 19 in a sequence table, (2) an anti-CD3 antibody labeled with a fluorescent dye A, (3) an anti-gamma-interferon antibody labeled with a fluorescent dye B, and (4) a Golgi apparatus blocking agent, wherein the fluorescent dye A and the fluorescent dye B emit different fluorescent colors. A novel method which can be used for detecting mycobacterium tuberculosis infection is provided. As proved by experiments, antigenic stimulation can be performed by directly using the peripheral blood without separating single karyocytes of the peripheral blood. As indicated by experimental data, the kit has high sensitivity and high specificity when being used for detecting mycobacterium tuberculosis infection. Moreover, the kit is easy and convenient to operate and low in cost, and has high clinical application value.

The invention discloses a human immunocompetent cell DC-CIK cell preparation and an effective preparation method of the human immunocompetent cell DC-CIK cell preparation. The method includes the following steps: (1) isolating single mononuclear cell from the human peripheral blood; (2) inoculating the single mononuclear cell into the serum-free medium, adding IFN gamma, GM-CSF and IL-4 and culturing for 1 day; (3) adding IL-2, IL-15 and anti-CD3 monoclonal antibody and culturing for 2 days; (4) adding TNF alpha, continuously culturing for 1day and promoting the DC cell maturation; (5) adding IL-2 and IL-15, continuously culturing for 8 days and obtaining the DC-CIK cell; and (6) collecting the DC-CIK cell and preparing the cell preparation. Through the culturing program of the DC-CIK cell, the human immunocompetent cell DC-CIK cell preparation not only can shorten the cell culturing time, improve the multiplication rate and strengthen the killing activity of the DC-CIK cell on the cancer cell, but also can simplify the operating steps and facilitate the clinic popularization and application.

The invention discloses a culture medium for activating, amplifying and culturing NK (Natural Killer) cells in vitro, a method for activating, amplifying and culturing the NK cells in vitro by utilizing the culture medium, and a biological agent containing the NK cells which are prepared through the method and are activated and amplified in vitro. Mononuclear cells are re-suspended by using the special culture medium for the NK cells, and a human CD3 antibody and a TLR7/8 agonist are added. The special culture medium for the NK cells is a serum-free medium containing human albumin, IL-2, IL-15 and in-vitro human plasma.