8 results about "Jurkat cells" patented technology

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 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 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 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 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.

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.