54 results about "T reg cells" patented technology

Species of human-derived bacteria belonging to the Clostridia class have been shown to induce accumulation of regulatory T cells (Treg cells) in the colon and suppress immune functions. Pharmaceutical compositions containing these bacteria can be used to prevent and treat immune-mediated diseases such as autoimmune diseases.

The invention discloses a sorting and amplification method of human peripheral blood CD4+CD25+Foxp3+ regulatory T cells, and belongs to the field of cell sorting amplification. The sorting and amplification method of the human peripheral blood CD4+CD25+Foxp3+ regulatory T cells specifically comprises the following steps: (1) separating peripheral blood mononuclear cells; (2) performing MACS (magnetic activated cell sorting) on CD4+CD25+Foxp3+Treg cells; and (3) performing in-vitro culture and amplification on the CD4+CD25+Foxp3+Treg cells, and finally performing amplification and culture to obtain the human peripheral blood CD4+CD25+Foxp3+ regulatory T cells. According to the sorting and amplification method disclosed by the invention, human peripheral blood CD4+CD25+Treg cells are successfully separated by using an immuno-magnetic bead two-step process, and the purity of the human peripheral blood CD4+CD25+Treg cells is subjected to flow identification to ensure that the purity of the finally-obtained CD4+CD25+Treg cells can reach 97%, and the activity of the cells can reach more than 95%.

The invention belongs to the technical field of cell culture in vitro, and particularly relates to a preparation method of high-purity, high-multiplication capacity and high-cytotoxin activity CIK (cytokine induced kill) cell. The method comprises the following steps: collecting and separating peripheral blood mononuclear cell of a patient, eliminating CD4+CD25+Treg cell by means of Mini MACS (magnetic active cell sorting) method, and sorting to obtain CD3+, CD4+ and CD8+T cells; and putting the obtained cells into culture solution containing phytohemagglutinin (PHA), so that the PHA concentration in the suspension liquid is 100ng / ml, hatching for 24h under the culture condition of 5% CO2 at 37 DEG C, transferring the hatched suspension liquid into a cell culture bottle coated by CD3 monoclonal antibody (1mug / ml), adding IFN (interferon)-gamma (1000U / ml), adding IL (interleukin)-2(500U / ml) and IL (interleukin)-21(1000U / ml) after 48h, compensating sodium selenite-containing (0.005mg / L)cell culture after four days, and continuously culturing for 7-14 days to obtain the high-purity, high-multiplication capacity and high-cytotoxin activity CIK (cytokine induced kill) cell. The quantity, the activity and the purity of the CIK cell which is prepared by the method and amplified in vitro are improved, so that the antineoplastic function of the CIK is enhanced.

An affinity ligand is reactive to the GARP protein may be capable of binding to an extracellular domain of GARP protein expressed on regulatory T (Treg) cells. The affinity ligand may be an antibody and may be used to identify Treg cells. A method comprises providing a blood sample from a subject and determining the amount of Treg cells in that sample. A composition containing Treg cells may be administered to an individual to suppress effector T cell activity in the individual. A composition containing an affinity ligand capable of binding to a GARP domain may be administered to an individual to suppress Treg cell activity and increase effector T cell activity in the individual. A kit for detecting Treg cells may include an affinity ligand reactive with mammalian GARP protein.

The present invention provides nanoparticle-coupled tolerogenic Treg cell therapy for treatment of immune and / or autoimmune disorders. In certain specific embodiments, the present invention can be used in the prevention and / or treatment of autoimmune diseases including, but not limited to, type 1 diabetes, lupus erythematosus (SLE), multiple sclerosis (MS), inflammatory bowel disease (IBD), rheumatoid arthritis, oophoritis, and autoimmune pathology associated with Graft versus Host Disease (GvHD) following hematopoietic stem cell transplantation.

The invention discloses a method for detecting an immunosuppression function of human regulatory T cells. The method comprises the following steps: collecting peripheral blood for extracting a mononuclear cell; using fluorescein marked anti-CD4 and anti-CD25 antibodies for marking a cell surface antigen; using a fixing membrane-breaking agent for fixing the cell surface antigen; after breaking the membrane, using fluorescein marked anti-Foxp3 and anti-Helios antibodies for marking endonuclear transcription factor Foxp3 and Helios; and finally, using a flow cytometry for detecting CD4+CD25+FoxP3+Helios+Treg cell population. According to the method, the function of the regulatory T cells in the peripheral blood is directly analyzed in the manner of quickly detecting the expression condition of the children peripheral blood transcription factor Helios. According to the method, the anti-Helios, anti-CD4, anti-CD25 and anti-Foxp3 flow antibodies can be used for more quickly and accurately marking the peripheral blood Treg cells with higher immunosuppression function.

A method of identifying an antigen-specific regulatory T cell (Treg) from a subject is discussed wherein the method comprises quantitatively or qualitatively detecting co-expression of each of cell markers CD4, CD25 and CD134, or alternatively, N each of cell markers CD8, CD25 and CD137, as well as one or more cell markers selected from the group of Treg cell markers consisting of CD39, CD73, CD127, CTLA-4 and Foxp3 on a cell in a suitable lymphocyte-containing sample from the subject in response to exposure to a target antigen. Also discussed are methods of isolating and expanding the identified antigen-specific Treg population, which may permit antigen-specific Treg cell therapy.

The invention discloses application of ginsenoside Rh2 and particularly verifies that ginsenoside can be applied to preparation of inhibitors for inhibiting Treg cells or preparation of compositions for preventing and / or treating tumors with Treg cells increased. The ginsenoside Rh2 is capable of improving balance of T cells in human bodies by inhibition of the Treg cells, autoimmunity can be further enhanced, and accordingly an anti-tumor effect or an immunodeficiency disease improvement effect can be achieved.

The present invention provides methods and compositions for expanding Treg cells ex vivo or in vivo using one or more conjugates comprising a costimulatory moiety that stimulates at least one of three signals involved in Treg cell development and / or using dendritic cells pulsed with antigens and modified to display TGF-β, or hematopoetic stem cells or bone marrow cells modified to display TGF-β. The methods and compositions are useful, for example, in the treatment and prevention of autoimmune disease, including Type 1 diabetes and in preventing foreign graft rejection, as well as to establish mixed chimerism, induce tolerance to autoantigens, alloantigens or xenoantigens, beta cell regeneration, prevention of foreign graft rejection, and treatment of a genetically inherited hematopoietic disorder.

The invention discloses application of L-fucose in medicines and health-care products for treating gastrointestinal lesions. The L-fucose is used for treating the gastrointestinal lesions, such as inflammatory bowel disease, functional gastrointestinal disorder and irritable bowel syndrome. The medicine is a carbohydrate medicine, since the adverse effect is less, and the tolerance of a patient is good, so that the medicine can be widely applied into clinic. The medicine has the advantages that the L-fucose is used in an early period of the gastrointestinal lesions, the activation of mesenteric lymph node Treg cells is regulated by DC cells, the differentiation potential of colons Th1 and Th17 is inhibited, the generation of bile acid is inhibited by regulating the intestinal flora, the function of inhibiting the contraction and spasm of intestinal muscles is realized by an nNOS path, the intestinal inflammation and weight-losing conditions are improved, and the patient can obtain an obvious long-term prognosis benefit; because of limitations of related existing therapy medicines, namely multiple medicine types, large side effect and poor compliance, a new therapy strategy for treating the patients with the gastrointestinal lesions, such as the inflammatory bowel disease, the functional gastrointestinal disorder and the irritable bowel syndrome, is provided.

The invention discloses lactobacillus casei capable of relieving rheumatoid arthritis and application of the lactobacillus casei, and belongs to the technical field of microorganisms. A lactobacilluscasei CCFM1074 strain is screened out by the invention, and the lactobacillus casei CCFM1074 has the effect of relieving rheumatoid arthritis. The effect is specifically reflected in the following aspects: a weight loss of a rat suffering from rheumatoid arthritis is remarkably relieved; content of pro-inflammatory cytokines TNF alpha and IL-1 beta in serum of the rat with rheumatoid arthritis isremarkably reduced; content of short-chain fatty acid in excrement of the rat suffering from rheumatoid arthritis is remarkably adjusted; ratios of Th17 cells and Treg cells in mesenteric lymph nodesof the rat with rheumatoid arthritis are remarkably reduced; intestinal flora of the rat with rheumatoid arthritis is improved at a genus level; and the lactobacillus casei CCFM1074 obviously inhibitsthe growth of pathogenic bacteria Prevotella copri in vitro.

The invention relates to the technical field of cell culture, in particular to a culture method for improving peripheral blood CTL (cytotoxic lymphocyte) cells. The method includes the steps: 1) inoculating PBMC separated by peripheral blood into a culture vessel coated by a CD3 antibody and a CD28 antibody to cultural the PBMC; 2) adding IL-1 alpha and IL-2 with the final concentration of 900U / mL-1100U / mL, IL-7 with the concentration of 40ng / mL-60ng / mL and IL-12 with the concentration of 15ng / mL-25ng / mL into a culture system; 3) exchanging liquid. A cell factor in the culture system is IFN-gamma with the concentration of 900U / mL-1100U / mL, and a cell factor in a new culture system is IL-2 with the concentration of 900U / mL-1100U / mL. According to the method, CTL amplification effects are good, activated CTL proportion is higher, and the proportion of T-reg cells is lower.

Provided is a use of miRNA-214 inhibitor in inhibiting regulatory T cells (Treg cells). The microRNA-214 (miRNA-214) can promote Treg cells, and can assist tumours in immune escape. The experiments demonstrate that the inhibition of miRNA-214 can inhibit the growth of Treg cells, thus inhibiting the growth of tumours. Therefore, miRNA-214 can be used for developing an anti-tumour drug or a drug inhibiting immune response hyperactivity.

The invention relates to an in-vitro amplification method of cord blood Treg cells, comprising the steps of S1, isolating mononuclear cells; S2, adjusting cellular density, inoculating to a culture flask, and adding inactivated plasma; on the first day, adding coumestrol, TGF-beta, CD3 / CD28 monoclonal antibody and IL-2; S3, on the fourth day, supplementing a culture medium, adding inactivated plasma, coumestrol and TGF-beta, and keeping the final concentration constant; S4, on the sixth day, supplementing the culture medium, and keeping the final concentrations of other components unchanged; S5, on the ninth day, supplementing the culture medium, and keeping the final concentrations of other components unchanged; S6, on the twelfth day, collecting cells. The amplification of CD4, CD25 and Tregs is induced under the coaction of coumestrol and TGF-beta, the amplifying efficiency is high, and high-purity Treg cells can be acquired without pre-purification.

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.

The present invention provides materials and methods related to culturing stable Treg cells or in vivo expansion of stable Treg cells, the cells made by the methods, treatments for various inflammatory / autoimmune pathologies and transplant / graft rejection, and related materials. Ex vivo induction and expansion of the stable Tregs is described, including use of inducing compositions, such as certain mAbs and other compounds, along with expansion medium comprising IL-2. In vivo expansion of stable Treg cells and treatments for various inflammatory / autoimmune pathologies and transplant / graft rejection are described, including the use of mAbs and their variants.

The invention relates to a culture solution of universal Treg cells and a culture method and application of the culture solution. The culture method of the culture solution of the universal Treg cellscomprises the following steps: S110, co-culturing CD4+ CD25+ Treg cells and hUC-MSCs according to a certain proportion for 48 h-72 h, and centrifuging to obtain supernatant; S220, preparing peripheral blood PBMC and plasma, inactivating the plasma, and centrifuging for later use; S330, adding cytokine, calcium ionophore, plasma obtained from S220 and a RPMI1640 culture solution into the supernatant obtained from S110 to prepare a Treg cell culture solution; S440, inoculating peripheral blood PBMC into the Treg cell culture solution prepared in S330 according to a certain density, and culturing for 48 h-96 h; S550, collecting cells, wherein the cell viability is greater than 90%, the cell expression of CD4+ CD25+ Foxp3+ Treg is greater than 65%, and CD4+ CD25+ Foxp3+ Helios+ Treg cells account for more than 90% of the cell expression of CD4+ CD25+ Foxp3+ Treg. The problems that Treg cell culture operation is complex and the period is too long are solved, and the problems that the content of CD4+ CD25+ Foxp3+ Helios+ Treg cells in peripheral blood is too low and Treg cell treatment cannot be implemented due to a patient with low quality of autologous Treg cells caused by the problemof diseases of the patient are solved.

The invention relates to a culture method of peripheral blood CTL cells. The method comprises the following steps: 1) inoculating PBMC obtained by separating from placental blood to a culture vessel coated with a CD3 antibody and a CD28 antibody for culture, wherein the cell factor in the culture system is IFN-gamma 900U / mL-1100U / mL; 2) adding IL-1alpha and IL-2 into the culture system, with the final concentrations being both 900U / mL-1100U / mL, and also adding IL-7 into the culture system, with the final concentration being 4-6ng / mL; and 3) changing the solution, wherein cell factors in the new culture system are IL-2 900U / mL-1100U / mL, IL-7, 2-3ng / mL, and the basal culture medium is a VIVO culture medium containing 4-6% autologous plasma. The method has the advantages that the CTL amplification effect is good, the activated CTL proportion is relatively high, and the T-reg cell proportion is relatively low.

The invention relates to an in vitro isolation and amplification method and an application of human peripheral blood Treg cells, and belongs to the technical fields of in vitro isolation and amplification methods and applications of cells. The method comprises the following steps: 1) collecting peripheral blood of the mate of an unexplained recurrent spontaneous abortion patient, and separating peripheral blood mononuclear cells by gradient centrifugation; 2) washing the peripheral blood mononuclear cells twice with a phosphate buffer solution, and counting the obtained mononuclear cell; 3) separating CD4<+>T cells and Treg cells by a magnetic bead sorting technology; and 4) amplifying the Treg cells. The method allows the Treg cells with a high purity to be obtained, the number of cells,required for clinical application, is obtained by the in vitro amplification method, and the human peripheral blood Treg cells obtained in the invention can be used for treating the unexplained recurrent spontaneous abortion, and can significantly improve the pregnant rate of unexplained recurrent miscarriage patients.

The present invention provides an immunomodulatory agent for use in the local treatment of tumours, wherein the treatment comprises patient-specific optimisation of the dose of the immunomodulatory agent to identify the maximum therapeutic dose that does not induce an increase in the number of local regulatory T cells (Treg cells) in the patient The invention further provides methods for the local treatment of tumours as well as methods for optimising treatments for the same

The invention discloses a traditional Chinese medicine composition for regulating chronic hepatitis B. The traditional Chinese medicine composition comprises the following components in parts by weight: 10-20 parts of raw astragalus, 5-15 parts of radix sophorae flavescentis, 10-20 parts of fried Chinese yams, 5-15 parts of glossy privet fruits, 5-15 parts of semen cuscutae, 5-15 parts of rhizoma smilacis glabrae, 5-15 parts of radix curcumae, 10-20 parts of polygonum cuspidatum, 10-20 parts of underleaf pearls and 5-15 parts of salvia miltiorrhiza. The invention also provides application of the composition to preparation of a drug for treating chronic hepatitis B and regulating peripheral blood Treg cells. Concretely, the invention provides a decoction prepared according to the traditional Chinese medicine formula and application of the decoction to preparation of a drug for treating chronic hepatitis B by matching with a western drug. As a whole, the traditional Chinese medicine composition prepared according to the whole formula has the effects of tonifying the spleen and benefiting the kidneys as well as removing heat from the liver and dispersing blood stasis; and when being united with the western drug for treating chronic hepatitis B, the traditional Chinese medicine composition can be used for effectively regulating the peripheral blood Treg cells and is beneficial to the treatment of chronic hepatitis B.

The invention relates a method for inducing in-vitro expansion of CD8<+> regulatory T cells by immunosuppressants. According to the method, expansion of CD8<+> regulatory T cells is jointly induced by TGF-beta and RAPA (rapamycin). The invention further provides a CD8<+> Treg (regulatory T cell) with an immunosuppression function and application of the CD8<+> Treg with the immunosuppression function. The method has the advantages that the TGF-beta and the RAPA are added into a polycloning in-vitro expansion system of the CD8<+> regulatory T cells for the first time, and a great number of CD8<+> Tregs with the remarkable immunosuppression function are obtained successfully; as multi-round expansion is conducted, the CD8<+> Tregs can increase exponentially so as to meet the demands of clinical cell treatment. The CD8<+> Tregs can be kept stable in the aspects of activity, purity, phenotype, nullipotency, suppression function and the like and can be used for treating various autoimmune diseases through adoptive infusion, thereby being huge in clinical application potential.

An affinity ligand is reactive to the GARP protein may be capable of binding to an extracellular domain of GARP protein expressed on regulatory T (Treg) cells. The affinity ligand may be an antibody and may be used to identify Treg cells. A method comprises providing a blood sample from a subject and determining the amount of Treg cells in that sample. A composition containing Treg cells may be administered to an individual to suppress effector T cell activity in the individual. A composition containing an affinity ligand capable of binding to a GARP domain may be administered to an individual to suppress Treg cell activity and increase effector T cell activity in the individual. A kit for detecting Treg cells may include an affinity ligand reactive with mammalian GARP protein.

The invention relates to the field of medicine and discloses application of vitamin D in resisting Aspergillus fumigatus. Compared with the prior art, the invention discloses, for the first time, application of vitamin D in resisting Aspergillus fumigatus; in-vivo experiments show that vitamin D can inhibit activity of conidia of Aspergillus fumigatus and decrease the amount of in-vivo Treg cells;in-vitro experiments show that vitamin D can inhibit the formation of autophagic lysosomes so as to resist infection of Aspergillus fumigatus; in-vivo and in-vitro experiments verify that vitamin D can maintain effective autophagy level and the certain amount of Treg cells in early stage of Aspergillus fumigatus removal in lungs so as to prevent damage from excessive autophagy; vitamin D enhancesthe cellular immunity of a host and can, after the infection of Aspergillus fumigatus, adjust autophagy and Treg cells to fight against the infection of Aspergillus fumigatus.

The invention belongs to the technical field of cell culture in vitro, and particularly relates to a preparation method of high-purity, high-multiplication capacity and high-cytotoxin activity CIK (cytokine induced kill) cell. The method comprises the following steps: collecting and separating peripheral blood mononuclear cell of a patient, eliminating CD4+CD25+Treg cell by means of Mini MACS (magnetic active cell sorting) method, and sorting to obtain CD3+, CD4+ and CD8+T cells; and putting the obtained cells into culture solution containing phytohemagglutinin (PHA), so that the PHA concentration in the suspension liquid is 100ng / ml, hatching for 24h under the culture condition of 5% CO2 at 37 DEG C, transferring the hatched suspension liquid into a cell culture bottle coated by CD3 monoclonal antibody (1mug / ml), adding IFN (interferon)-gamma (1000U / ml), adding IL (interleukin)-2(500U / ml) and IL (interleukin)-21(1000U / ml) after 48h, compensating sodium selenite-containing (0.005mg / L)cell culture after four days, and continuously culturing for 7-14 days to obtain the high-purity, high-multiplication capacity and high-cytotoxin activity CIK (cytokine induced kill) cell. The quantity, the activity and the purity of the CIK cell which is prepared by the method and amplified in vitro are improved, so that the antineoplastic function of the CIK is enhanced.

The present invention relates to a composition comprising an isolated CD4+ Treg cell population, wherein the Treg cells have signatures for i) identifying that the T-cells are CD4+ regulatory Tcells, ii) identifying that the Treg cells are tissue type tropic, i.e they can migrate to the diseased tissue, iii) optionally identifying that the Treg cells are tropic with respect to the diseased tissue, i.e. they are homing cells, iv) identifying that the Treg cells are emigrant cells, i.e. they originate from the target tissue, and v) optionally identifying that the Treg cells are retained in the target tissue and optionally one or more X-signatures and / or one or more Y-signatures and one ore more Z-signatures.

The present invention relates to composition comprising an isolated CD8+ Treg cell population, wherein the Treg cells have signatures for i) identifying that the T-cells are CD8+ regulatory Tcells, ii) identifying that the Treg cells are tissue type tropic, i.e they can migrate to the diseased tissue, iii) optionally identifying that the Treg cells are tropic with respect to the diseased tissue, i.e. they are homing cells, iv) identifying that the Treg cells are emigrant cells, i.e. they originate from the target tissue, and v) optionally identifying that the Treg cells are capable of being retained in the target tissue and optionally one or more X-signatures and / or one or more Y-signatures.