17 results about "Jurkat cells" patented technology

The invention discloses an application of a DUF3109 protein in preparation of an antitumor drug. The DUF3109 protein is a bacteroides ovatus DUF3109 protein. The DUF3109 protein can promote the formation of a three-level lymph structure in a tumor by regulating CXCL13 secreted by CD4 + T cells, so that the progress of liver cancer is inhibited. In a specific embodiment, the DUF3109 protein shows that the DUF3109 protein has a remarkable inhibition effect on liver in-situ tumors and remarkably promotes CD4 < + > T lymphocytes and Jurkat cells to secrete CXCL13. According to the technical scheme, the application of the DUF3109 protein is expanded, a potential innovative solution is provided for tumor treatment, and a new research direction is provided for development of anti-tumor drugs.

The invention belongs to the technical field of cell culture, and particularly relates to a method for preparing Jurkat cells on a large scale by utilizing a bioreactor. The preparation method comprises the following steps: (1) cell resuscitation: resuscitating cells from liquid nitrogen; (2) carrying out cell multiplication culture: carrying out step-by-step amplification multiplication culture in T bottles and cell culture bags with different specifications; (3) large-scale culture of cells: performing large-scale culture by using a bioreactor; and (4) harvesting the cells: collecting the cell suspension, and centrifugally harvesting the cells. The preparation method overcomes the problem that a large number of thymus cells or human lymphocytes need to be used as immunogens when the anti-human T lymphocyte immune globulin is prepared, can simply and quickly prepare a large number of Jurkat cells, is used for preparing the anti-human T lymphocyte immune globulin, improves the quality, reduces the biological safety risk, and has a wide application prospect. The method is beneficial to industrial production and has a relatively good economic effect.

The invention belongs to the field of gene engineering, and particularly relates to application of chikungunya virus structural protein in improvement of pseudovirus stability, a construction method of the chikungunya virus structural protein and vaccines, antibody evaluation and in-vivo gene therapy products. The method comprises the following steps: constructing a human codon optimized Asian strain CHIKV-E3 + E2 + 6K + E1 structural protein eukaryotic vector, and designing two types of binder expression plasmids of targeted T cells; the vector, a psPAX2 helper plasmid and a lentivirus target vector containing an SFFV promoter and a ZsGreen reporter gene (or a CAR therapeutic gene) are co-transformed into a 293T cell, and the lentivirus is prepared through culture, filtration and centrifugation. The lentivirus can efficiently infect Jurkat cells and activate human T cells, obviously reduces the infection efficiency on 293T cells so as to reduce the off-target effect, has excellent stability in human serum, and provides a safe and efficient gene delivery tool for in-vivo CAR-T therapy.

The present invention provides the use of isogaminol compounds in combination with glucocorticoids in the preparation of a leukemia treatment drug, belonging to the technical fields of biology and medicine. The isogaminol compounds described herein, when used in combination with glucocorticoids, can enhance the sensitivity of Jurkat cells to glucocorticoids, induce G1 cell cycle arrest in Jurkat cells, and induce apoptosis in Jurkat cells. The combined use of isogaminol compounds and glucocorticoids in the present invention provides a research foundation for overcoming glucocorticoid resistance in acute lymphoblastic leukemia.

The present invention relates to a method for inducing and expanding natural killer cells derived from peripheral blood mononuclear cells, which comprises co-culturing, as feeder cells, irradiated Jurkat cells and irradiated Epstein-Barr virus transformed lymphocyte continuous line (EBV-LCL) cells in the presence of cytokines, along with peripheral blood mononuclear cells. According to the present invention, a large quantity of natural killer cells can be induced and proliferated from a small quantity of peripheral blood mononuclear cells even without the use of high-cost equipment or various kinds of expensive cytokines, thereby making it possible to significantly improve the efficiency and efficacy of the prevention and treatment of cancer using the natural killer cells.

The present invention relates to CD3e / g targeting antibodies or antigen binding fragments thereof, their preparation and uses. The invention discloses a CD3e / g targeting antibody or an antigen binding fragment thereof, the antibody comprises VH and VL, the VH comprises VH CDR1 shown as any one amino acid sequence of SEQ ID NO: 1-6, VH CDR2 shown as any one amino acid sequence of SEQ ID NO: 7-15 and VH CDR3 shown as any one amino acid sequence of SEQ ID NO: 16-22; the VL comprises a VL CDR1 shown as an amino acid sequence of any one of SEQ ID NO: 23 to 27, a VL CDR2 shown as an amino acid sequence of SEQ ID NO: 28 or 29, and a VL CDR3 shown as an amino acid sequence of any one of SEQ ID NO: 30 to 32. The antibody can be combined with human or monkey CD3e / g, and can effectively activate an NFAT downstream signal channel of a Jurkat cell.

The application relates to application of a CaMKII-alpha inhibitor in preparation of a medicine for treating T cell lymphoma. The application can obviously down-regulate the expression level of CaMKII-alpha in T cell lymphoma through si-RNA, determines that CaMKII-alpha is a potential treatment target of T cell lymphoma, and identifies Evans blue as an effective CaMKII-alpha inhibitor through high-throughput screening. Experiments show that the Evans blue can obviously inhibit the growth of T cell lymphoma Jurkat cells and H9 cells, and the effect is similar to that of siRNA treatment. In-vivo experiments of Jurkat transplanted tumors also prove that the Evans blue realizes the in-vivo tumor inhibition effect by inhibiting CaMKII-alpha, and reveal a good tumor inhibition effect of the Evans blue on T cell lymphoma.

The invention discloses a reporter gene cell line based on an IL2 promoter and construction and application of the reporter gene cell line, and belongs to the technical field of biology. The reporter gene cell line based on the IL2 promoter is obtained by inserting multiple copies of NFAT, AP-1 and NF-kappa B nucleoprotein binding site genes into the upstream of a modified miniaml IL2 promoter, connecting a modified luciferase reporter gene to the downstream of the modified miniaml IL2 promoter and packaging and infecting Jurkat cells through lentivirus. Further, the expression quantity of luciferase reporter genes in the reporter gene cell line based on the IL2 promoter is detected, so that the activation of the T cells is efficiently and quickly detected; besides, the IL2 promoter-based reporter gene cell line can be used for screening drugs for targeting T cell pathway activation or preparing a model for monitoring IL2 promoter activation and / or inhibition, so that the IL2 promoter-based reporter gene cell line has a relatively good application prospect.

The invention relates to the technical field of immunotherapy, and discloses a TCR mRNA rapid synthesis method and application thereof, a TCR double-stranded template is rapidly synthesized through two rounds of PCR amplification by adopting a mode of complementary CDR3 + J region primer + fixed fragment '5' UTR + V region 'and' C region + 3 'UTR' fragment, and then TCR mRNA is synthesized through IVT. According to the application, the reliability of the method is confirmed through first-generation sequencing, TCR specific screening is carried out by co-transforming alpha / beta double-stranded mRNA into Jurkat cells, and finally, the TCR recognition capability is further verified by co-transforming alpha / beta double-stranded mRNA into human primary T cells. The synthesis period is short, vector construction is avoided, time cost is saved, TCR synthesis flexibility is improved, and high-throughput screening of specific TCRs can be achieved in a short time.

This invention discloses the application of Huanglian Shangqing tablets combined with glucocorticoids in the preparation of drugs for treating leukemia. The Huanglian Shangqing tablets combined with glucocorticoids can inhibit the activity of Jurkat cells in a concentration-dependent manner; inhibit the proliferation of Jurkat cells in a concentration- and time-dependent manner; and significantly induce apoptosis in the human acute lymphoblastic leukemia cell line Jurkat. In animal experiments, compared with the model group, the Huanglian Shangqing tablets combined with glucocorticoids can increase the hematocrit ratio of leukemia mice, reduce the spleen burden in mice, have no obvious toxic side effects, and improve the survival rate of leukemia mice. Therefore, the Huanglian Shangqing tablets combined with glucocorticoids have the potential value of developing drugs to reverse glucocorticoid resistance in leukemia. Furthermore, Huanglian Shangqing tablets are already used clinically, which can effectively shorten the preclinical research period and has a very good application prospect.

The invention discloses an application of tautogen in preparation of an anti-cancer drug. Studies show that the Taiwanin has an obvious anti-cancer effect, research data with Jurkat cells, Hela cells and A549 cells as objects show that the Taiwanin has an obvious inhibition effect on leukemia cells, cervical cancer cells and lung cancer cells, the Taiwanin has the value of preparing the anti-cancer drugs, and the Taiwanin is derived from natural plants, is relatively high in safety and has a good application prospect. The compound has great value in the aspect of developing and preparing anti-cancer drugs. The invention not only provides a new choice for anti-cancer drugs, but also opens up a new direction for the utilization of taiwania flousiana plant resources.

According to the invention, the chemiluminescence and electrochemical dual-mode sensor based on the magnetic biological aggregate is innovated through the electrostatic interaction between DNA and Fe3O4-NH2 nanoparticles and the catalytic reduction capability of the Fe3O4-NH2 nanoparticles to H2O2. The method comprises the following steps: firstly, synthesizing Fe3O4-NH2 by adopting a hydrothermal method, and then combining the Fe3O4-NH2 with TdT extended DNA to form a sensor core; the sensor can generate chemiluminiscence and electrochemical signals through catalytic reaction when H2O2 exists, and the intensity of the chemiluminiscence and electrochemical signals changes along with the change of the concentration of H2O2 or TdT. The TdT inhibitor PP is added to limit the TdT activity and reduce the signal strength. The technology realizes efficient detection of H2O2, TdT and PP, is applied to quantitative analysis of TdT in Jurkat cells, and shows the application potential in biomedicine and clinical diagnosis. The method is simple to operate, low in cost and accurate in detection.

The invention belongs to the technical field of biology, and discloses application of a 5 '-UTR sequence in improvement of RNA translation efficiency and a nucleic acid molecule. The invention provides application of a 5 '-UTR sequence as shown in any one or more of nucleotide sequences of SEQ ID NO: 1-51 in improvement of the translation efficiency of RNA (Ribonucleic Acid). The 5 '-UTR sequence provided by the invention is introduced into a linear mRNA and / or circular RNA structure, so that the translation efficiency of the 5'-UTR sequence in human cells such as HEK293 cells, HepG2 cells, Jurkat cells and 293A cells can be effectively improved, and the 5 '-UTR sequence shows huge application potential in the aspect of optimizing the treatment effect of RNA molecules such as linear mRNA and circular RNA and has a good application prospect.

The invention belongs to the technical field of cell biology, and particularly relates to a triple co-culture cell model for immune function research as well as a construction method and application of the triple co-culture cell model. Based on HMEC-1, THP-1 and Jurkat cells, a triple co-culture cell model is constructed by adopting a Transwell three-dimensional construction mode, and a microenvironment in which T cells exist in human blood can be simulated. The obtained triple co-culture cell model is subjected to toxic attack by using mixed benzene, and the expression of CD3 on the surface of the Jurkat cell is not destroyed by co-culture when contamination is carried out for 24-48 hours, so that the TCR signal transduction function is reserved. After contamination is carried out for 24 hours, the expression of ICAM-1 of Jurkat cells of the triple co-culture cell model is up-regulated, and the regulation of an in-vivo immune microenvironment on the activation state of the T cells can be better reflected. The sensitivity of the triple co-culture cell model is higher than that of single culture, and the immune function can be better researched.

The present invention belongs to the field of drug detection and specifically discloses a method for detecting the antibody-dependent cell-mediated cytotoxic biological activity of human immunoglobulin, comprising the following steps: 1) taking a target cell suspension and adding it to a well plate, then adding a series of concentrations of human immunoglobulin solutions for incubation, and finally adding an effector cell suspension for incubation; 2) taking the well plate incubated in step 1) to detect fluorescence, draw a dose-effect curve and calculate the half-maximal effective concentration; the target cells are CHO-K1 cells expressing the SARS-CoV-2 Spike protein, and the effector cells are Jurkat cells stably expressing CD16a on the cell membrane and stably transfected with the firefly luciferase gene driven by the NFAT response element. The detection method of the present invention can accurately and reliably detect the antibody-dependent cell-mediated cytotoxic biological activity of two different human immunoglobulin preparations, IVIG and SCIG.

A method for simulating sensitive response of Jurkat cells in mechanical microenvironments, includes the following steps: S1, transfecting FRET probes into the Jurkat cells by using an electroporation method; S2, simulating the Jurkat cells in the mechanical microenvironments for a first time; S3, collecting FRET images of the Jurkat cells by using a FRET microscope; S4, performing quantitative analysis and statistical analysis on the FRET images; S5, simulating the Jurkat cells in the mechanical microenvironment for a second time; and S6, detecting an ERK phosphorylation level by using a Western blot method. By simulating different states of Jurkat cells, such as no-coated adhesion, charge adsorption, extracellular matrix adsorption, and endothelial cell layer adhesion, and by using methods of FRET living cell observation and biochemical detection method of ERK phosphorylation antibody, the method detects the Jurkat cells in different states, to obtain ERK activity of Jurkat cells in different mechanical microenvironments.

The present invention discloses the use of 1,3-dicyclohexylbarbituric acid combined with dexamethasone in the preparation of a drug for treating leukemia. Experimental testing shows that 1,3-dicyclohexylbarbituric acid can increase the sensitivity of leukemia to dexamethasone. In vitro testing using the glucocorticoid-resistant acute T lymphocytic leukemia cell line Jurkat reveals that 1,3-dicyclohexylbarbituric acid and dexamethasone have a significant synergistic effect on Jurkat cells, and 1,3-dicyclohexylbarbituric acid can enhance the sensitivity of Jurkat cells to dexamethasone in a concentration- and time-dependent manner. The combination of 1,3-dicyclohexylbarbituric acid and dexamethasone can induce G1 arrest in Jurkat cells and regulate GR / BIM and JAK2 / STAT3 signal transduction to activate the caspase cascade reaction, thereby inducing Jurkat cell apoptosis. The 1,3-dicyclohexylbarbituric acid of the present invention has the potential to become a dexamethasone sensitizer, and also provides a research basis for the development of drugs to overcome glucocorticoid resistance.