36 results about "Cytokine-induced killer cell" patented technology

The invention discloses a preparation method of human cytokine-induced killer cells, comprising the following steps: coating a cell culture flask with a coating buffer containing effective amount of fusion protein and human CD3 monoclonal antibody before culturing precursor cells of human CIK cells, and adding the human CD3 monoclonal antibody in the whole process of inducing and culturing the human CIK cells, wherein the fusion protein is human intercellular adhesion molecule-1 functional domain and human fibronectin functional domain fusion protein, and the concentration of the human CD3 monoclonal antibody in the cell culture solution is lower than the concentration of the human CD3 monoclonal antibody in the coating buffer. According to the invention, ex-vivo expansion efficiency of peripheral blood mononuclear cells and the proportion of CD3/CD56 double positive cells in the CIK cells are significantly raised, the cytotoxicity activity of the CIK cells is enhanced, thus the effect of cellular immunity treatment is raised.

The invention discloses an efficient CIK amplifying method, in particular to a method for cell populations, namely cytokine-induced killer cells utilizing in-vitro cell factors for efficiently inducing mononuclear cell expressions CD3 and CD56 of peripheral blood, wherein the cell populations have killing functions. The cultivation method for the CIK efficiently expressing CD3+CD56 comprises the following steps that the peripheral blood of a healthy person or a patient is collected in a sterile mode, after the peripheral blood is diluted through a saline solution with the same volume, a Ficoll lymphocyte separating medium is used for separating mononuclear cells, in the CIK cell inducing process, CD3 monoclonal antibodies (CD3mAb), CD28 monoclonal antibodies (CD28mAb), interferon-gamma (IFN-gamma), interleukin-2(IL-2) and interleukin 1alpha (IL-1alpha) are added, cultivation is carried out for 13-16 days to obtain cells, a CIK cell preparation is prepared, and flow cytometry detection and microorganism detection are carried out. According to the CIK cultivation method, the CIK number can be increased to be 6*10<9> or over 6*10<9> in two weeks, the cell survival rate can be increased to be 99% or over 99%, and the double-positive proportion of the CD3+CD56+cells reaches 30% or over 30%.

The invention discloses a method for stimulating the rapid proliferation of CIK (Cytokine-induced Killer) cells by using DC (Dendritic Cells). The method comprises the following steps: separating peripheral blood monouclear cells (PBMC); performing induction culture on cytokine-induced killer cells, i.e., performing induction culture on the CIK cells; performing induction culture on the DC; and detecting the phenotype CD3<+>CD56<+> of the CIK cells. According to the method, the numbers of the CIK cells are amplified substantially within a short period while the CD3<+>CD56<+> ratio of the surfaces of the CIK cells is not changed sharply, so that the amplification time of the CIK is shortened obviously; and the proliferation rate is improved at the same time. Therefore, the important scientific research value is obtained.

The invention discloses an immunity enhancing reagent comprising AAVs carrying molecule encoding genes of antibodies at immunodetection points, wherein the immunodetection points are one or several from PD-1, PD-L1 and CTLA-4. The AAVs carrying the molecule encoding genes of the antibodies at different immunodetection points are combined with each other, T cells, natural killer cells, cytotoxic T lymphocytes or cell factor induced killer cells and other immune cells are infected in vitro, and the antibodies at the immunodetection points are expressed and secreted after the infected immune cells are fed back to bodies of patients suffering from tumors, so that the combination of receptors at the immunodetection points and ligands of the receptors is blocked, the transfer of an immunosuppression signal is effectively blocked, the immune tumor suppression microenviroment is destroyed, the immune cell killing capacity is improved, and furthermore the curative effect of adoptive immune therapy is improved.

A mass production of dendritic cells (DC) and cytokine-induced killer cells (D-CIK) cells from the umbilical cord blood or peripheral blood via the activation with a cellular activator “Koyo” derived from the purified tumor cells during the aggregation and proliferation of the purified tumor cells is provided. A preparation of a vaccine with the dendritic cells (DC) and D-CIK cells is used to improve the symptoms of infectious diseases, cancers, metastasised cancers and auto-immune disorders.

The invention discloses a CIK (Cytokine-Induced Killer Cell) capable of knocking down endogenous PD-1 (programmed death 1) expression and a preparation method and application thereof. The invention provides a method for preparing a CIK preparation capable of knocking down endogenous PD-1 expression, comprising the following steps: A) inducing the differentiation of PBMC to CIK in vitro by using cytokines; B) transferring a vector capable of knocking down endogenous PD-1 expression into the CIK obtained in step A), culturing for 13 to 16 days and harvesting a cell, thereby obtaining the CIK preparation capable of knocking down the endogenous PD-1 expression, wherein the vector capable of knocking down endogenous PD-1 expression contains a plurality of tandem encoded genes targeting miRNAs of PD-1, and the number of the encoded genes is 2 to 30. Experimental results show that the vector can significantly reduce the expression level of endogenous PD 1 in the CIK, thereby significantly enhancing the tumor-killing activity of a CIK cell.

The invention relates to an application of dendritic cells combined with cytokine-induced killer cells in lung cancer treatment. The invention prepares sensitized DC cells and CIK cells; the two cells are transfused back to patients with lung cancer, which can effectively prolong the survival time of the patients; and the toxic and side effect is low.

The invention belongs to the field of biomedicine and discloses application of DC-CIK cells in preparation of a medicine used for resisting HIV infection. Through a lot of experiments, the inventor discovers that when CIK cells impacted by HIV and DC cells are co-cultured, the CIK cells can obviously promote maturation of the DC cells. When the DC and CIK cells are combined for use, the DC and CIK cells have mutual promotion presentation and a function for killing cells infected by HIV. After 30 days of the DC-CIK cell infusion, the number of CD4T cells of a HIV infector is obviously increased, the killing function of NK (Natural Killer) and CIK cells is enhanced, and HIV viral load is obviously restrained. Thus, kill cells induced by co-cultured dendritic cells and cytokines can be applied to the preparation of the medicine used for resisting HIV infection.

The invention provides a dendritic cell, a preparation method thereof and application. Concretely, the invention provides application of combination of the dendritic cell and cytokine-induced killer cell to prepare a medicine for treating or inhibiting liver cancer, wherein the dendritic cell is capable of specifically recognizing liver cancer nidus. According to the embodiment in the invention, the combination of the dendritic cell and the cytokine-induced killer cell is capable of effectively treating or inhibiting liver cancer.

The invention provides a method for intensifying functions of cytokine-induced killer cells. The method comprises the following steps: analyzing and obtaining an epitope through a bioinformatics analysis method, fusing the epitope and Ii-Kye into an epitope peptide, anchoring the epitope peptide on MHC-II molecules on the surfaces of dendritic cells, and infecting the dendritic cells with recombinant adenovirus rAd-RNAi-SOCS1; and further carrying out co-cultivation on the dendritic cells and cytokine-induced killer cells, and treating tumor-bearing mice with co-cultivated cells, so that the processed dendritic cells are capable of activating the cytokine-induced killer cells, and intensifying the significant killing functions of the cytokine-induced killer cells on related tumors. The invention provides a method for treating tumors employing activated cells; and the core technology of the method has reference significance on treatment of other viroses.

The invention relates to applications of BCG-PSN (BCG-polysaccharide nuceic acid). The BCG-PSN is capable of promoting proliferation of CIK cells (cytokine-induced killer cells) and capable of improving an anti-tumor effect of the CIK cells; the expression of related cytotoxic activity factors is also enhanced when the anti-tumor efficiency is improved; and the CIK cells, which accept induction culture of the BCG-PSN, are applicable to cell therapy of tumor diseases.

The invention discloses a frozen resuscitation solution for multiple cytokine induced killer cells, and an application thereof. The frozen resuscitation solution for multiple cytokine induced killer cells comprises albumin and dextran. The multiple cytokine induced killer cells obtained through the subsequent induction and amplification process of single karyocytes obtained through resuscitation using the frozen resuscitation solution have the advantages of fast proliferation rate, high induced differentiation efficiency and good cell viability.

The invention discloses an immune preparation for treating tumor; the immune preparation contains dendritic cells (DC) loaded with a tumor antigen and cytokine-induced killer cells (CIK), the concentration of the DC loaded with the tumor antigen in a culture liquid is 1*10<7> to 1*10<8>, and the concentration of the CIK in the culture liquid is 1*10<9> to 1*10<10>. The invention further provides a preparing method of the immune preparation for treating tumor; the preparing method comprises the steps: collection of mononuclear cells of peripheral blood; isolation and culture of the DC and the CIK; extraction of a tumor antigen gene and viral vector packaging; transfection of the DC with a virus mediated tumor antigen gene; and co-culture of the DC loaded with the tumor antigen and the CIK. The accuracy of positioning of re-transported immune cells in a human body can be improved. The objective defect of a conventional cell immunotherapy technology in the same field is not good in solid tumor treatment effect can be effectively improved; adaptation diseases are wide, and the use is safe and effective.

The invention provides a cytokine-induced killer cell activity-associated protein and an application thereof, relates to an immune cell activity-associated protein and especially relates to influences of haptoglobin on variation of cell activity of cytokine-induced killer (CIK) cells. The expression of the haptoglobin at a protein level and a nucleic acid level is significantly increased with enhancement of the activity of the CIK cells, which means that a positive correlativity exists between the haptoglobin and the activity of the cytokine-induced killer cells. The invention provides an index of detection of an activity intensity of the CIK cells, or provides basis for researching a recombinant protein and a relative medicine which are used for improving an immune system with the haptoglobin as an active component.

The invention belongs to the technical field of biotechnology, and particularly relates to a DC (Dendritic cell)-CIK (Cytokine-induced killer cell) immune cell modified by both AFP (Alpha-fetoprotein)and HBsAg (Hepatitis B surface antigen) antigen genes as well as a preparation method and application of the cell. The preparation method provided by the invention comprises the following steps: firstly constructing a recombinant plasmid pHAGE-AFP-HBsAg; then the recombinant plasmid pHAGE-AFP-HBsAg, a helper plasmid psPAX2 and pMD2G being co-transfected to 293T cells with calcium phosphate transfection reagent, to obtain recombinant lentivirus AFP-HBsAg; subsequently, infecting DC cells with the recombinant lentivirus AFP-HBsAg, to obtain the DC cells modified by the AFP-HBsAg; and finally, co-culturing the DC cells modified by the AFP-HBsAg with the CIK cells, to obtain the DC-CIK immune cells.

The invention relates to a cell cryopreservation solution for cryopreservation of immune cells and a cryopreservation method thereof, wherein killer cells induced by cytokines are particularly good. The cell cryopreservation liquid does not contain serum and is prepared from the following components: (1) a mixed basic culture medium, (2) dimethyl sulfoxide (DMSO), (3) a serum substitute and (4) dextran. The serum-free cryopreservation liquid is characterized in that the serum-free cryopreservation liquid is a mixed culture medium and a serum substitute so as to achieve a serum-free cryopreservation function. After immune cells, especially natural killer cells, are unfrozen, 80% or more of the cell poisoning function and functional cell markers can be maintained, and high-concentration cell loading capacity is achieved.

The present invention relates to activated lymphocytes comprising cytokine-induced killer cells in which CD8⁺CD56⁺NKG2D⁺ cells are present at a proportion of 20% or more, and a preparation method therefor, and more particularly, to activated lymphocytes comprising cytokine-induced killing cells which have high tumor cell killing abilities and growth rates and are almost free of side effects because they do not require the combined administration of interleukin-2, and a preparation method therefor.

The invention discloses a preparation method of cytokine-induced killer cells. The method comprises the following steps of: separating a collected human peripheral blood sample to obtain PBMNC; inoculating the PBMNC into a culture flask coated with a recombinant humanized anti-CD25mAb monoclonal antibody for culture; and transferring non-adherent cells obtained in the step S2 and culture supernatant into a culture flask coated with a recombinant humanized anti-CD3 epsilon monoclonal antibody, and supplementing INF-gamma for continuous culture. According to the preparation method of the cytokine-induced killer cells, CIK cells are induced and amplified by adopting a traditional culture process of CD25 monoclonal antibody improved CIK cells, so that the yield of CD3+ CD56+ cells is increased, the proportion of CD3+ CD4+ cells is reduced, the killing potential of the CIK cells on K562 and SK-MEL-5 cells is effectively improved, and the preparation method has important significance for formulating a preparation method and quality standard of a clinical-grade CIK cell preparation.