306 results about "Peripheral blood mononuclear cell" patented technology

The invention discloses a pharmaceutical composition for promoting aTreg cell differentiation and application thereof, and relates to the field of biological medicine, and the pharmaceutical composition comprises zinc salt, a basic cell culture medium, fetal calf serum, antibiotic, L-glutamine and a pH buffer agent. According to the application, the basic cell culture medium is used for providing basic nutrition required by cell growth for lymphocytes, the fetal calf serum is used for providing growth factors, hormones, adhesion proteins and nutrient substances required by cell growth for the lymphocytes, and the antibiotics are used for preventing bacterial infection; l-glutamine provides key amino acid for cell energy metabolism and participates in synthesis of protein and nucleic acid, zinc salt provides zinc ions, and under the comprehensive action, the proportion of Treg cells and aTreg cells differentiated from lymphocytes in peripheral blood mononuclear cells is increased.

The invention belongs to the field of new application of medicines, and particularly relates to application of SIRT4-IN-1 in preparation of medicines for treating autoimmune diseases. It is found for the first time that SIRT4-IN-1 reduces the activation degree of immune response induced by HSV-1 virus infection by up-regulating acetylation modification of cGAS, inhibits abnormal activation of immune response in Trex1 deficient mouse cells and peripheral blood mononuclear cells of systemic lupus erythematosus patients, and improves abnormal activation of autoimmune response. The invention provides a new candidate drug for clinical treatment of systemic lupus erythematosus.

The invention belongs to the technical field of biomedicine, and particularly relates to application of GPR161 as an acute distress syndrome diagnostic marker and a therapeutic target. Experiments prove that the expression level of GPR161 in peripheral blood mononuclear cells of a patient with the acute respiratory distress syndrome is remarkably increased, and the expression quantity of the GPR161 is positively correlated with the severity of the disease, so that the GPR161 gene or GPR161 protein can be used as a molecular marker for screening or diagnosis or prognosis evaluation of the acute respiratory distress syndrome; a reagent for inhibiting GPR161 gene expression or protein activity can be used for preparing a medicine for treating the acute respiratory distress syndrome. The invention provides a new strategy for diagnosis, monitoring and targeted drug development of ARDS, and has important clinical application value.

The invention discloses a monoluciferase-based ADCP biological activity detection method, which comprises the following steps: separating a mononuclear cell from a human peripheral blood mononuclear cell as an effector cell, phagocytizing a Raji cell transfected by Luciferase as a target cell by the mononuclear cell, acidifying and degrading the Raji cell in a phagocytic body, and degrading the Luciferase so as to attenuate a light-emitting signal, so that the ADCP biological activity is detected. The ADCP biological activity of the antibody therapeutic drug is accurately quantified by detecting the weakening of the light-emitting signal. According to the present invention, the method only needs to separate the mononuclear cells and perform Luciferase infection on the target cells, and only needs to add the luciferase substrate after the phagocytosis process, such that the luminescence value can be read by using the microplate reader, the operation process is simple, the steps are less, the time consumption is short, the high-throughput screening is achieved, and the new direction is provided for the ADCP detection.

The invention provides a preparation method and a kit for reprogramming induced pluripotent stem cells of PBMC (peripheral blood mononuclear cells), particularly provides PBMC reprogramming based on mRNA-LNP (peripheral blood mononuclear cells), and more particularly relates to a preparation method for reprogramming induced pluripotent stem cells of peripheral blood mononuclear cells. The preparation method comprises the following steps: preparing a nano liposome mRNA (messenger ribonucleic acid) cocktail-LNP; according to the method, the nano-liposome mRNA-LNP is used for inducing peripheral blood mononuclear cells to reprogram, so that reprogrammed induced pluripotent stem cells are obtained, and an iPSCs reprogramming technology which is low in cost, low in toxicity, efficient, stable, repeatable and safer is realized. The invention also provides a reprogramming kit and a preparation method of the induced pluripotent stem cells.

The invention relates to a three-level lymphoid structure model and a preparation method thereof, and belongs to the technical field of cell biology. The method for preparing the three-stage lymphoid structure model comprises the following steps: S1, inducing and activating peripheral blood mononuclear cells by using an extracellular matrix to obtain activated peripheral blood mononuclear cells; s2, the peripheral blood mononuclear cells and the mesenchymal stem cells activated in the step S1 are coated with hydrogel, and hydrogel microspheres are obtained; and S3, culturing T cells and B cells by using the hydrogel microspheres obtained in the step S2 to obtain the three-stage lymphatic structure. The three-level lymphoid structure model obtained by the invention maintains long-term stability in vitro, the form is normal, and the activity of T cells and B cells is strong. The method can be used for dynamically observing the three-level lymphoid structure in the subsequent research immunotherapy and drug screening process, is used for high-throughput experiments, effectively improves the efficiency of drug screening experiments, and is high in accuracy.

The present invention relates to an immunoprophylactic and therapeutic vaccine comprising a peripheral blood mononuclear cell loaded with a natural killer T cell ligand and an antigen, and more particularly, to an immunotherapeutic vaccine comprising a peripheral blood mononuclear cell loaded with alpha-galactosylceramide, a natural killer T cell ligand and a glycolipid. The compositions of the present invention are readily available because there is no need to isolate specific cells from peripheral blood mononuclear cells. Besides, the immunization of the peripheral blood mononuclear cells loaded with the natural killer T cell ligands and antigens not only induces the obvious-level activation of the natural killer cells and the natural killer T cells and the response of cytotoxic T lymphocytes, but also has a synergistic treatment effect in the aspect of treating malignant tumors. Therefore, the compound can be beneficially used as an anti-cancer immunotherapeutic agent.

The invention relates to an application of a small molecule medicine STK000311 in a medicine for treating a cGAS-STING pathway abnormal activation disease, and belongs to the technical field of biological medicines. According to the application disclosed by the invention, the small-molecular medicine STK000311 or the pharmaceutically acceptable salt of the small-molecular medicine STK000311 serving as the cGAS-STING signaling pathway inhibitor has an obvious treatment effect on the cGAS-STING pathway abnormal activation diseases for the first time, and verification experiments show that the small-molecular medicine STK000311 can be used for obviously inhibiting the activity of human cGAS protein in a targeted manner and inhibiting cGAS-mediated antiviral immune response; the transcription level of ISGs in PBMCs (peripheral blood mononuclear cells) of an SLE (systemic lupus erythematosus) patient is inhibited, the fact that the small-molecule medicine STK000311 has huge potential of developing cGAS activity related diseases is fully proved, a novel treatment medicine is provided for clinic, and the small-molecule medicine STK000311 has good clinical application value and wide application prospect.

The invention belongs to the technical field of gene engineering, and relates to an esophageal squamous cell carcinoma diagnostic kit based on a peripheral blood mononuclear cell methylation marker and application. The kit comprises a primer pair which is used for detecting the methylation of a cg05914150 site, a cg06769875 site, a cg07475126 site, a cg08433285 site, a cg10038907 site, a cg17968322 site, a cg19572487 site, a cg20184330 site, a cg22741248 site and a cg22823192 site. According to the detection kit provided by the invention, the type of a detected sample is peripheral blood, the sample is relatively easy to obtain and non-invasive, and the patient compliance is relatively good. Abnormal methylation changes mostly exist in the early progression process of tumors, early diagnosis of ESCC can be achieved through the methylation detection method provided by the invention, and the five-year survival rate of ESCC patients is effectively increased.

The invention belongs to the technical field of immunology, and particularly discloses an in-vitro preparation method and application of an induced follicular regulatory T cell iTfr.The in-vitro preparation method comprises the steps that mouse spleen single-cell suspension or healthy human peripheral blood is obtained, peripheral blood mononuclear cells are obtained, then initial CD4 + T cells are obtained through immunomagnetic bead sorting, and iTfr cells are obtained; then carrying out induced differentiation by using an antibody and a specific cytokine, and carrying out continuous stimulation and culture by adopting two different cytokine polarization conditions, so as to obtain the iTfr cell. The invention provides a method for preparing the iTfr cells in vitro, which is expected to obtain an enough number of induced follicular regulatory T cells, and provides an important tool and a treatment means for in vitro research of immune tolerance related diseases such as autoimmune diseases and organ transplantation.

The invention belongs to the field of biological medicine, and particularly relates to a CD3-LILRB4 bispecific antibody and application thereof. The invention provides a novel bispecific antibody with high activity and low toxicity, the bispecific antibody can be combined with CD3 and LILRB4 so as to promote T cell activation and effectively promote PBMC (peripheral blood mononuclear cells) to kill tumor cells, the bispecific antibody has good anti-cancer activity, and the bispecific antibody has the performance of reducing secretion of proinflammatory cytokines, is higher in safety, and can be applied to preparation of anti-tumor drugs. Good clinical application and drug development values are realized.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-CD38 nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing CD38 antibody protein, immunizing alpaca, obtaining immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the cells, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), carrying out PCR (polymerase chain reaction) to obtain antibody gene segments, constructing the antibody gene segments on a phage expression vector to construct a CD38 immune antibody library, carrying out two rounds of panning, carrying out Clone ELISA (enzyme-linked immuno sorbent assay) detection on clones to obtain an antibody combined with the CD38 protein, carrying out sequencing analysis, and selecting a unique sequence. High-throughput expression is carried out through a mammalian cell high-throughput expression system, antibody protein is obtained, FACS detection is carried out through a CD38 high-expression cell line, and finally the antibody combined with CD38 is obtained. The method is simple and rapid to operate, a mammalian cell expression system induces efficient expression of the antibody, processing modification can be performed after translation, and the activity of the antibody is closer to that of a natural antibody.

The invention relates to a construction method of a nanometer antibody coding DNA library. The method comprises the following steps: screening animal peripheral blood mononuclear cells (PBMC) by using immunogen to obtain positive cells specifically bound with the immunogen; and amplifying a nucleic acid sample derived from the positive cells so as to obtain a coding DNA library of the nano-antibody, wherein the nano antibody is a nano antibody specifically binding to the immunogen, and wherein the amplification comprises the use of one or more groups of primer pairs, each group of primer pairs of the one or more groups of primer pairs comprising one or more forward primers and one or more reverse primers. A large number of diversified DNA sequences covering a large number of low-abundance sequences are obtained. According to the technical route for rapidly preparing the VHH antibody based on high-throughput sequencing, the high-affinity and high-specificity VHH antibody can be efficiently and rapidly obtained, and the raw material requirement of a high-quality diagnostic reagent is met.

The invention provides a method for obtaining GiNK cells through novel culture medium amplification, a large number of highly purified human natural killer (NK) cells are obtained by combining IL-15 active fragments, PI3K inhibitors, key receptors targeting the surfaces of the NK cells, antibodies enhancing activation of the NK cells and various cytokines, and the GiNK cells can be used for preparing the human natural killer cells. Expansion from human peripheral blood mononuclear cells (PBMCs) can be achieved without the use of feeder cells. Through culture for 2-3 weeks, the GiNK cells can be amplified by hundreds to thousands of times, and the proliferation efficiency, functional activity and anti-tumor effect of the GiNK cells are remarkably improved. The culture medium provided by the invention not only reduces the use concentration of cell factors, but also solves the problem of function depletion of GiNK cells in in-vitro culture through the synergistic effect of various additives, and provides important technical support for clinical application of the GiNK cells.

The invention discloses a preparation method and application of a chimeric antigen receptor NK cell, and relates to the technical field of immunization, and the preparation method comprises the following steps: (1) recovery of a peripheral blood mononuclear cell (PBMC); (2) sorting NK cells; (3) carrying out multiplication culture on the sorted NK cells; (4) adding cell factors into the NK cells subjected to multiplication culture, and performing activation culture; the cell factor is a combination of IL-2 (Interleukin-2), IL-15 and IL-21 (Interleukin-21); and (5) adding Polybrene into the activated and cultured NK cells, infecting for 2-24 hours by using lentivirus carrying CAR molecules and having MOI of 1-10 virus amount according to the cell density of 2 * 10 < 5 >-1 * 10 < 6 > / mL, changing the liquid after infection, and continuously culturing for 54-72 hours to obtain the chimeric antigen receptor NK cells. According to the invention, the optimized conditions are utilized to prepare the CAR-NK cells, and the CAR positive rate can reach 93.87%.

The invention provides an ischemic stroke auxiliary diagnosis system based on a peripheral blood T cell single cell transcriptome and application thereof, and the auxiliary diagnosis system comprises a peripheral blood mononuclear cell acquisition module, a single cell RNA sequencing module, a data processing module, a data analysis module and a result discrimination module. With a coding gene of a protein molecule significantly related to the occurrence of ischemic stroke as a target gene, a data processing module obtains the expression level of the target gene in each T cell; the data analysis module obtains a risk score of each T cell of the subject by using a built-in single cell risk scoring model, and weights the risk scores of all the T cells through distributed perception identification to obtain an individual risk score of the subject; and the result judgment module is used for classifying the subjects into ischemic stroke negative and ischemic stroke positive according to the individualized risk scores of the subjects. The auxiliary diagnosis system disclosed by the invention is high in ischemic stroke discrimination capability, and high in sensitivity and specificity.

The invention relates to the field of traditional Chinese medicine, in particular to a construction method for an individualized traditional Chinese medicine intelligent prescription evaluation model, a medicine evaluation method, a processor and electronic equipment or device. The curative effect of different traditional Chinese medicine combinations on the patient is evaluated by reversing the degree of a peripheral blood mononuclear cell imbalance pathway of the patient through the traditional Chinese medicine combinations. According to the method, the traditional Chinese medicine composition is evaluated in a targeted manner or a personalized intelligent formula is prepared on the basis of the signal path enrichment condition of the peripheral blood mononuclear cells of the patient in the current disease state. According to different disease states of patients, targeted existing traditional Chinese medicine compositions are dynamically screened in real time or new effective prescriptions are intelligently designed. According to the method, a quantification method is adopted, and real-time and dynamic composition and evaluation of the traditional Chinese medicine or the composition are achieved according to the disease state of a patient.

A method of assessing the safety of a gene therapy vector comprising: providing a population of genetically modified cells 18 wherein the cells have been transfected with a vector; exposing the genetically edited cells to a carcinogen 20; growing the cells to produce immortal cell lines; and comparing the survival of the test cells with a control 14 to determine if the vector contributes to carcinogenicity. Also claimed is a method of testing the safety of a vector comprising: providing test cells; genetically editing the cells with a vector and exposing cells to a carcinogen or exposing the cells to a carcinogen and modifying the cells with a vector; growing the cells; and comparing the survival of the cells to a control. The cells may be used in immunotherapy. The cells may be CAR-T cells. The cells may be from peripheral blood mononuclear cells or induced pluripotent stem cells (iPSCs). The carcinogen may be ENUA. Also claimed is a method of designing a vector for use in gene therapy by harvesting nucleic acid from the test cells, characterising the modification of the test cells, redesigning the vector, and repeating the method of testing the safety of the vector.

The invention discloses an application of Notch2 overexpression lentivirus in promoting aTreg cell differentiation, and relates to the field of immune cell culture, and the application comprises the following steps: suspending PBMC in a T cell culture medium, laying a plate, adding Notch2 overexpression lentivirus, and culturing at 36.8-37.2 DEG C. The invention also discloses a preparation method of the Notch2 overexpression lentivirus. According to the application, the Notch2 overexpression lentivirus and the PBMC (peripheral blood mononuclear cell) are co-cultured on a culture medium, and the PBMC (peripheral blood mononuclear cell) is transfected by utilizing the Notch2 overexpression lentivirus, so that Notch2 protein molecules are overexpressed, and the number of Treg cells and aTreg cells in lymphocytes is increased.

The present invention discloses a method for evaluating the anti-tumor activity of immune cells in vitro and its application. The present invention first constructs a co-culture model: (1) peripheral blood mononuclear cells are separated from the peripheral blood of healthy people, and γδT cells are obtained after pre-stimulation and amplification culture; (2) tumor tissue is digested using a tumor tissue dissociation kit Tumor Dissociation Kit, and then the digested tumor cells are cultured for organoids, and tumor organoids of 37μm to 100μm are obtained after screening; (3) Corning 96-well all-white flat-bottom polystyrene microplates are pre-coated with anti-adhesion reagents, and then tumor organoids are plated, counted, and then mixed with γδT cells to form a co-culture system to obtain the co-culture model; finally, the co-culture model is further used to evaluate the anti-tumor activity and evaluate the killing effect of γδT cells on tumors.

The present disclosure provides methods and kits for determining antigen-specific T cell responses in a subject based on the use of a whole blood sample rather than isolated, enriched, and / or extracted peripheral blood mononuclear cells. The use of a whole blood sample to measure antigen-specific T cell responses in a subject streamlines the method by reducing sample preparation and consumption. These methods for determining antigen-specific T cell responses in a subject include contacting a whole blood sample from the subject with at least one antigen to form a mixture, contacting the whole blood sample with at least one staining reagent, and analyzing the mixture for antigen-specific T cells.

An apparatus and method for T-cell separation, activation, transduction and expansion. Three-dimensional (3D) bioreactors may be employed that include antibody coatings. Such 3D bioreactors can be employed for T-cell separation from peripheral blood mononuclear cells including attachment of T-cells to the 3D bioreactor surface for activation and transduction by lentivirus vectors to produce CAR T-cells. The CAR T-cells can then be expanded in a separate downstream bioreactor therein providing a scalable automated system.

The present disclosure describes PLPC-DB, an ultrapure phospholipoproteomic composition consisting of essential phospholipids, bioactive proteins, and intercellular regulatory factors, derived from the supernatant of peripheral blood mononuclear cells (PBMCs). This composition achieves a purity level exceeding 99% through a patented purification process that integrates high-speed advanced centrifugation and selective ultrafiltration, ensuring the structural stability and functional integrity of its bioactive components. PLPC-DB is optimized for advanced research and diagnostic applications, providing reproducibility, consistency, and safety across multicenter studies. The essential biomolecular components of PLPC-DB include phosphatidylcholine and phosphatidylserine, which contribute to membrane stability and intracellular signaling, while cell communication peptides enhance intercellular signaling, homeostatic regulation, and biochemical coordination. Structural and regulatory lipids support cell membrane biogenesis and functional stability, whereas adhesion and signaling proteins mediate cell-cell interactions and immune response coordination. Additionally, bioactive regulatory factors modulate immune and inflammatory responses, contributing to tissue regeneration and metabolic homeostasis.

The present disclosure belongs to the field of biotechnology, and specifically relates to a preparation method for an adenovirus p53 (Ad-p53)-loaded dendritic cell (DC) vaccine. The present disclosure includes steps of peripheral blood collection and peripheral blood mononuclear cell (PBMC) separation, PBMC sorting, DC activation, Ad-P53-transfected DC and DC vaccine preparation. P53 can be expressed on a surface of DC as a tumor-associated antigen (TAA) through DC purification, specific multiplicity of infection (MOI) and infection modes, and the Ad-P53-transfected DC has obvious antigen presentation effect, which can be used as a vaccine to activate T cells to kill tumors.

The invention discloses a method for constructing astrocytes serving as a smoke disease model, and belongs to the technical field of crossing of stem cells and neuroscience. The method comprises the following steps: S1) reprogramming CD34 + cells in in-vitro PBMCs (peripheral blood mononuclear cells) of smoke disease patients carrying and not carrying RNF213p.R4810K mutation to obtain induced pluripotent stem cells; and S2) directionally inducing and differentiating the induced pluripotent stem cells into astrocytes through a neural progenitor cell way, wherein the obtained astrocytes are the astrocytes capable of being used as the smoke disease model. The astrocyte model prepared by the invention can be used for researching pathogenesis, nerve-blood vessel interaction process and blood-brain barrier (BBB) related functions of smoke diseases, and can be further applied to molecular typing of diseases and in-vitro function evaluation of candidate drugs.

The application discloses application of falcarinol-7-methyl ether and derivatives thereof in preparation of medicines and functional foods for delaying aging or immune regulation. In the application, the falcarinol-7-methyl ether and derivatives thereof are derived from falcarinol-7-methyl ether, and can effectively delay replicative senescence of mesenchymal stem cells, inhibit expression of aging-related markers of mouse liver and lung induced by adriamycin, improve grip strength of the mouse, enhance muscle function, and promote proliferation of T cells separated from human peripheral blood mononuclear cells, and have an immune regulation effect. Therefore, the falcarinol-7-methyl ether and derivatives thereof have good effects of improving telomerase activity, anti-aging and immune regulation, and can be developed into anti-aging or immune regulation products.

The invention discloses an application of a circRNA (Ribonucleic Acid) marker in preparation of a reagent for diagnosing rheumatoid arthritis, the circRNA marker is hsacircRNA001715, and the expression of the hsacircRNA001715 in peripheral blood mononuclear cells of a cold-dampness arthralgia type rheumatoid arthritis patient (active stage and inactive stage) is obviously up-regulated compared with that of a healthy control person; in addition, the hsacircRNA001715 can be used for up-regulating the expression of a peripheral blood mononuclear cell NLRP3 of a patient with the cold-dampness arthralgia type RA in the active period through an hsacircRNA001715 / miR-1249-5p axis and promoting an inflammatory reaction; the invention further discloses a kit for rheumatoid arthritis diagnosis, the kit can be rapidly used for early diagnosis of rheumatoid arthritis by detecting the expression level of hsacircRNA001715, operation is easy and convenient, accuracy is high, and a new strategy can be provided for clinical diagnosis of rheumatoid arthritis.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-NKG2D nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing NKG2D protein, immunizing alpaca to obtain immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the PBMC, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), obtaining an antibody gene segment through PCR (polymerase chain reaction), and constructing the antibody gene segment on a phage expression vector; the method comprises the following steps: carrying out electrotransformation on a vector with an antibody gene segment to an SS320 electrotransformation competent state to obtain an NKG2D immune antibody library, carrying out two rounds of NKG2D protein panning and Clone ELISA detection to obtain an antibody combined with NKG2D protein, carrying out sequencing analysis, selecting a unique sequence, and carrying out high-throughput expression through a mammalian cell high-throughput expression system to obtain an antibody protein. And carrying out FACS detection by using an NKG2D high-expression cell line to finally obtain the antibody combined with the NKG2D. The method disclosed by the invention is simple and quick to operate; a mammalian cell expression system induces high-efficiency expression of the antibody, processing modification can be performed after translation, and the activity of the antibody is closer to that of a natural antibody.

The invention belongs to the technical field of biomedicine, and particularly relates to a PAD-T cell targeting acute myelogenous leukemia cells and a preparation method and application of the PAD-T cell. The PAD-T cell comprises a T cell, a capsule shell coating the T cell, and a DNA aptamer connected with the capsule shell; wherein the capsule shell is formed by alternately overlapping a first polylysine layer, a first sodium alginate layer and a second polylysine layer, the DNA aptamer layer is an S30-T1 DNA aptamer, and the nucleotide sequence is as shown in SEQ ID NO. 1. According to the invention, the peripheral blood mononuclear cells of a healthy donor can be prepared on a large scale by utilizing a nano coating technology, and the recurrence after hematopoietic stem cell transplantation can be effectively relieved and the occurrence of GVHD can be reduced by combining an immune isolation concept and a biological material-nucleic acid material coated immune cell technology; the clinical application prospect is very high.

The application provides a construction method of a humanized keloid mouse model, comprising the following steps: BAC plasmid construction and preparation; superovulation of experimental mice and collection of zygotes; pronuclear microinjection of zygotes; post-injection embryo transplantation; genotype identification; breeding and genetic analysis; and verification of construction results of the humanized immune system by immunohistochemistry and immunofluorescence. In the application, peripheral blood mononuclear cells treated by sCD27 and skin around keloids (homologous cells and tissues) are transplanted into NSG-MHC-DKO immune-deficient mice with overexpression of CD70 genes, so that HLA rejection of different homologous immune cells and tissues can be avoided, the internal microenvironment of keloids and the interaction between the internal microenvironment and the immune system can be restored to the maximum extent, and the influence of immune factors on the occurrence and development of keloids can be realized in vitro. The application first discovers and verifies that the activation of the CD27-CD70 axis can promote the occurrence and development of keloids, and provides a suitable animal model for the research and development of anti-keloid drugs, especially immunotherapy.