46 results about "Naive T cell" patented technology

A new method to capture, purify and expand antigen-specific T lymphocytes has been developed using magnetic beads coated with recombinant MHC class I molecules. This method was optimized using homogenous populations of naive T cells purified from mice transgenic for the 2C T cell receptor (TCR). These T cells were captured on beads coated with MHC class I molecules and the relevant antigenic peptides. MHC and peptide specificity was confirmed by the usage of irrelevant MHC peptide combinations. An enrichment of 800 to 1600 fold was measured, using 2C T cells mixed with irrelevant T cells, starting from a 2C T cell frequency of 1 / 3000. The same approach was used to purify antigen-specific CD8+ T cells from total CD8+ T cells from naive mice. The recovered cells could be expanded and specifically kill target cells in vitro; they had a significant effect in vivo as well. We expect this procedure to be suitable to purify and expand in vitro tumor- and virus-specific killer T cells for use in cell therapy.

The invention belongs to the technical fields of immunology and tumor therapy, and relates to a preparation method and a use of a TCR gene modified CD8+T memory stem cell. The method comprises the following steps: co-incubating a tumor antigen and an immature dendritic cell to obtain a specific tumor antigen-supported mature dendritic cell, co-culturing the specific tumor antigen-supported maturedendritic cell and a CD8+ naive T cell, and adding a stem cell differentiation inhibitor to promote the generation of a CD8+ stem cell-like memory T lymphocyte; separating the CD8+ stem cell-like memory T lymphocyte; and cloning a T cell receptor gene (TCR) for specifically identifying a specific antigenic epitope, and carrying out lentivirus packaging or retroviral vector cotransfection on an autologous CD8+ Tscm cell to in-vitro prepare a CD8+ TCR-Tscm cell specific for different tumor specific antigens. The CD8+ TCR-Tscm cell prepared by the method has the advantages of overcoming of the tumor heterogeneity, high specificity, few adverse reactions, and highly-efficient and lasting tumor preventing and treating effects.

A method for overcoming mild to moderate immune suppression includes the steps of inducing production of naïve T-cells and restoring T-cell immunity. A method of vaccine immunotherapy includes the steps of inducing production of naïve T-cells and exposing the naïve T-cells to endogenous or exogenous antigens at an appropriate site. Additionally, a method for unblocking immunization at a regional lymph node includes the steps of promoting differentiation and maturation of immature dendritic cells at a regional lymph node and allowing presentation of processed peptides by resulting mature dendritic cells, thus, for example, exposing tumor peptides to T-cells to gain immunization of the T-cells. Further, a method of treating cancer and other persistent lesions includes the steps of administering an effective amount of a natural cytokine mixture as an adjuvant to endogenous or exogenous administered antigen to the cancer or other persistent lesions.

A method for overcoming immune suppression includes the steps of inducing production of naïve T cells and restoring T cell immunity. A method of vaccine immunotherapy includes the steps of inducing production of naïve T cells and exposing the naïve T cells to endogenous or exogenous antigens at an appropriate site. Additionally, a method for unblocking immunization at a regional lymph node includes the steps of promoting differentiation and maturation of immature dendritic cells at a regional lymph node and allowing presentation of processed peptides by resulting mature dendritic cells, thus, for example, exposing tumor peptides to T cells to gain immunization of the T cells. Further, a method of treating cancer and other persistent lesions includes the steps of administering an effective amount of a natural cytokine mixture as an adjuvant to endogenous or exogenous administered antigen to the cancer or other persistent lesions; preferably the natural cytokine mixture is administered in combination with thymosin α1.

A method for overcoming immune suppression includes the steps of inducing production of naïve T cells and restoring T cell immunity. A method of vaccine immunotherapy includes the steps of inducing production of naïve T cells and exposing the naïve T cells to endogenous or exogenous antigens at an appropriate site. Additionally, a method for unblocking immunization at a regional lymph node includes the steps of promoting differentiation and maturation of immature dendritic cells at a regional lymph node and allowing presentation of processed peptides by resulting mature dendritic cells, thus, for example, exposing tumor peptides to T cells to gain immunization of the T cells. Further, a method of treating cancer and other persistent lesions includes the steps of administering an effective amount of a natural cytokine mixture as an adjuvant to endogenous or exogenous administered antigen to the cancer or other persistent lesions; preferably the natural cytokine mixture is administered in combination with thymosin α1.

A new method to capture, purify and expand antigen-specific T lymphocytes has been developed using magnetic beads coated with recombinant MHC class I molecules. This method was optimized using homogenous populations of naive T cells purified from mice transgenic for the 2C T cell receptor (TCR). These T cells were captured on beads coated with MHC class I molecules and the relevant antigenic peptides. MHC and peptide specificity was confirmed by the usage of irrelevant MHC peptide combinations. An enrichment of 800 to 1600 fold was measured, using 2C T cells mixed with irrelevant T cells, starting from a 2C T cell frequency of {fraction (1 / 3000)}. The same approach was used to purify antigen-specific CD8+ T cells from total CD8+ T cells from naive mice. The recovered cells could be expanded and specifically kill target cells in vitro; they had a significant effect in vivo as well. We expect this procedure to be suitable to purify and expand in vitro tumor- and virus-specific killer T cells for use in cell therapy.

A method of purifying a subpopulation of naive T cells that have recently emigrated from the thymus, comprising (a) contacting a biological sample susceptible of containing T cells with at least four different ligands, namely ligands directed to the markers CD3, CD4, CD45RA, and CD31; (b) sorting the cells so as to recover T cells having a phenotype comprising at least the following four markers CD3+, CD4+, CD45RA+ and CD31hi, whereby a subpopulation of naive T cells is purified.

Novel antigen-presenting cells, including but not limited to dendritic cells, that are loaded with antigens from dead or dying cells including allogenic cell lines, and the methods for making such antigen-presenting cells are described. These loaded antigen-presenting cells induce therapeutic immune responses in humans. Such loaded antigen-presenting cells are useful in the management of cancer. Antigen-loaded dendritic cells prepared as described here can prime naïve T cells to differentiate into effector cells able to recognize multiple and / or shared tumor antigens that are expressed either on the tumor cells that are used to load the dendritic cells and / or on other tumor cells. The cytotoxic T cells generated by exposure to antigen-loaded dendritic cells prepared as described here can be used in adoptive therapy. This induction of responses against multiple antigens shared between different cells, for instance tumor cells, as described here is important as it leads to broad immune responses.

The invention provides a cell culturing method for improving the curative effect and lasting effect performance of chimeric antigen receptor T cells. By adding a tyrosine kinase inhibitor dasatinib (dasatinib), activation signal transmission of the CAR-T cells is reduced, the terminal differentiation of the CAR-T cells is inhibited, the proportion of naive T cells and central memory T cells in a CAR-T cell product is increased, and the depletion trend of the CAR-T cells is inhibited. The method solves the problems of the terminal differentiation and depletion trend of the CAR-T cells in the in-vitro culture process. The method is simple in culture process, easy to implement, low in cost and safe and reliable in clinic application; the proportion of the naive T cells and the central memoryT cells of the cultured CAR-T cells is high, the depletion trend of the CAR-T cells is avoided, and the good repeatability is achieved; the cultured CAR-T cells show the better curative effect and thebetter in-vivo lasting effect performance and have high application and popularization value.

A unique CD4+CD25+ regulatory T cell population develops from naive CD4+CD25− T cells during a TH1 polarized immune response (called TH1-TR cells). These TH1-TR cells can be generated by contacting naïve T cells with mature CD8α+ dendritic cells (DCs) that have been exposed to a TH1 polarizing adjuvant and, in some cases, an antigen of interest. The TH1-TR are identified by their expression of the cytokines IL-10 and IFN-γ, the transcriptional regulators T-bet and FoxP3, and the cell surface molecules CD4, CD25, CD69, CD44 and ICOS.

An object of the present invention is to provide a technique to distinguish between Treg cells and activated T cells in a live state. Another object of the present invention is to provide a pharmaceutical composition for immunostimulation that can reduce the number of Treg cells in vivo and effectively express the immune response of activated T cells. In the method of the present invention, Treg cells are detected from test cells containing (i) regulatory T cells and (ii) at least one type of cell selected from the group consisting of naive T cells and activated T cells, wherein expressions of folate receptor 4 on the surfaces of cells are measured and Treg cells are detected using the expressions as an indicator. The present invention uses anti-folate receptor 4 antibody or folate receptor 4-binding fragment as an active ingredient contained in a pharmaceutical composition for immunostimulation.

Flt3-ligand can be used to generate large numbers of dendritic cells from hematopoietic progenitor and stem cells. Flt3-ligand can be used to augment immune responses in vivo, and expand dendritic cells ex vivo. Such dendritic cells can then be used to present tumor, viral or other antigens to naive T cells, can be useful as vaccine adjuvants. When flt3-L is used and / or administered in combination with other reactive agents, e.g. CD40 binding proteins, 4-1BBL or antibodies reactive with 4-1BB, CD30 ligand antagonists, RANKL, and / or interferon alpha the combination further enhances immune responses and the effectiveness of vaccine adjuvants.

Approximately 50% of lymphocytic cells derived from peripheral blood is occupied by naive T cells before an activation culture is performed, but the ratio decreases through the activation culture. The occupancy of the memory T cells, which is less than half before the start of the culture, is increased by an activation culture to reach 95%, but decreases if the culture continues. An object of the present invention is to prepare CD45RO positive memory T cell groups in quantity without having to use an antibody or the like, conveniently, and rapidly, and to provide a pharmaceutical composition consisting mainly of the memory T cell groups prepared by the method. According to the present invention, harvested lymphocytic cells are activation-cultured, and the occupancy of the memory T cells in the lymphocyte cell group reaching a desired value is detected through the index of the density of the cultured lymphocyte cell group exceeding a predetermined threshold, the related cell group is harvested for free preservation, and the related free-preserved cell is thawed, cultured, and grown so as to produce the lymphocyte cell groups consisting mainly of the memory T cells in quantity.

Flt3-ligand can be used to generate large numbers of dendritic cells from hematopoietic progenitor and stem cells. Flt3-ligand can be used to augment immune responses in vivo, and expand dendritic cells ex vivo. Such dendritic cells can then be used to present tumor, viral or other antigens to naive T cells, can be useful as vaccine adjuvants. When flt3-L is used and / or administered in combination with other reactive agents, e.g. CD40 binding proteins and 4-1BBL or antibodies reactive with 4-1BB, the combination further enhances immune responses and the effectiveness of vaccine adjuvants.

The present invention is directed to generating suppressor cells by treating naive T cells with a suppressor-inducing composition such as anti-CD3, anti-CD28, IL-2, TGF-β, or some combination thereof. Such suppressor cells are administered to patients to prevent or treat immune disorders and are allogeneic to the patient.

The invention provides an application of vorinostat for preparation of immunomodulatory drugs. New properties of vorinostat are determined through a large quantity of experiments and include that vorinostat can inhibit STAT-1 and STAT-3 signal pathways, so that naive T cells are inhibited from differentiating to Th1 and Th17 lymphocytes, and IFN-gamma and IL-17 expression quantities are reduced; vorinostat can increase the number of nTreg cells of CD4<+>CD25<+>Foxp3<+>; vorinostat can inhibit expression of NF-kappa B P65, thereby inhibiting the expression quantity of TNF-alpha and relieving infiltration of macrophages; vorinostat can further increase the expression quantity of tight junction protein claudin-5, and accordingly, blood retinal barrier injury is repaired. On the basis of the properties, the novel application of vorinostat as the immunomodulatory drug is determined, the application of vorinostat used for experimental autoimmune uveitis therapeutic drugs is further determined, and the experiments prove that vorinostat has definite curative effect on experimental autoimmune uveitis, has a brand-new treatment pathway, is free of side effects and has a good application prospect.

The inventoin provides a use of ginsenoside F2 in the prevention, improvement or treatment of liver disease, and a pharmaceutical composition, a health functional food, and a feed composition including ginsenoside F2. Ginsenoside F2 inhibits fat synthesis and accumulation in the liver, and increases distribution of regulatory T cells capable of inhibiting activity of inflammatory cells, thereby preventing hepatitis, and also increases expression of anti-inflammatory cytokine IL-10 in regulatory T cells, and inhibits differentiation of naive T cells into Th17 cells, and is thereby effectively used for the treatment of various liver diseases.

A method for generating a purified regulatory T cell composition having a CD4+CD25+Foxp3− phenotype from naive T cells having CD4+CD45RA+RO−CD11alo CD27+ phenotype comprising the steps of stimulating a T cell receptor on the naive T cells and stimulating ICAM-1 on the naive T cells.

A method for generating a purified regulatory T cell composition having a CD4+CD25+Foxp3− phenotype from naive T cells having CD4+CD45RA+RO−CD11alo CD27+ phenotype comprising the steps of stimulating a T cell receptor on the naive T cells and stimulating ICAM-1 on the naive T cells.

The invention provides a kit for induced amplification of regulatory T cells from naive T cells. The kit comprises an anti-CD3 antibody, a PBS (phosphate buffer solution), a lymphocyte separation solution, a naive CD4<+>T Cell Biotin-Antibody Cocktail II antibody, Naive CD4[+]T Cell Micro Bead Cocktail II magnetic beads and a Treg induced culture medium. The invention also provides an application method of the kit. The kit and application method thereof are simple to operate, are safe and effective, and have the advantages of shorter amplification induction period, higher stability and lower cost.

Safe, rapid and efficient methods for producing antigen-specific T cells recognizing human papilloma virus or HPV antigens.

Disclosed are compositions derived from the commensal microbiota and related methods for inducing and differentiating naive T cells. In some embodiments, the compositions and methods can be selectively used to generate stable regulatory T-cells (Tregs or iTregs) or stabilize such Tregs to prevent inflammatory responses and instead promote antigen tolerance. Alternatively, in other aspects, select compositions can be used to promote pro-inflammatory T cell development, such as through the induced development and stabilization of Th1 and Th7 cells.

The invention discloses a method for evaluating the immunomodulatory function of a substance, which comprises the following steps: preparing a primary natural immune cell suspension from the spleen of an acquired immune cell deficient mouse; culturing the immune cells and OVA to obtain a culture system; collecting spleen and lymph node single-cell suspensions of a T cell receptor transgenic mouse, and carrying out magnetic bead sorting to obtain CD4 + CD62L + Naive T cells; adding the CD4 + CD62L + Naive T cells into the culture system, and adding a substance to be evaluated into the culture system at the concentration of 100-500mu g / mL in the culture process; collecting all cells, performing stimulation culture on the cells by using phorbol and ionomycin, performing membrane rupture, matching and fixing, adding a fluorescent antibody, and detecting the proportion of Th1, Th2 or Treg by using a flow cytometer so as to evaluate the immunomodulatory function of the substance. The method can accurately evaluate whether the active substance has the immune regulation function, and is low in cost and short in period.

The invention provides an optimized method for inducing amplification of regulatory T cells from naive T cells. The method comprises the following steps: (1) coating an anti-CD3 antibody; (2) isolating peripheral blood mononuclear cells; (3) sorting the naive T cells by virtue of magnetic beads; and (4) inducing amplification culture. The method provided by the invention is simple and convenient to operate, safe and effective, relatively short in amplification inducing cycle, relatively good in stability and low in cost.

Safe, rapid and efficient methods for producing antigen-specific T cells recognizing human papilloma virus or HPV antigens.