28 results about "Cell subpopulations" patented technology

The application provides a high-activity iPSC exosome and a preparation method and application thereof, relates to the technical field of biological medicine, and the preparation method comprises the following steps: obtaining induced pluripotent stem cells; performing flow cytometry sorting to obtain an iPSC subpopulation; performing suspension expansion culture on the iPSC subpopulation to collect a cell culture solution; sequentially performing clarification filtration, ultrafiltration concentration and CD9 antibody affinity chromatography purification, and then eluting and collecting an eluent containing exosomes. The induced pluripotent stem cells overcome donor dependence and batch difference. The specific screening of a cell subpopulation that is positive for CD105, CD73 and CD90 and maintains pluripotency eliminates heterogeneity and guarantees the high activity and uniform efficacy of the exosomes. The use of microcarriers for serum-free suspension culture breaks through the yield bottleneck and realizes large-scale production. Finally, the high-purity and high-activity exosomes are efficiently extracted through ultrafiltration and CD9 antibody affinity chromatography purification.

This application relates to the field of biotechnology, specifically disclosing the application of immune cell subsets in the preparation of detection products for assessing the efficacy of neoadjuvant therapy and the risk of recurrence and metastasis in breast cancer. The detection method described in this application is non-invasive and convenient, requiring only peripheral blood, and is easy to repeat sampling and dynamic monitoring. This application also provides a predictive model for the efficacy of neoadjuvant therapy in breast cancer, and a predictive model for the risk of recurrence and metastasis in breast cancer treated with neoadjuvant therapy. The constructed predictive model has a high AUC value (0.82-0.87), and good sensitivity and specificity. Furthermore, through regular postoperative monitoring, abnormal dynamic changes in immune cell subsets can be detected, potentially indicating signs of recurrence earlier than imaging and traditional tumor markers. This helps identify advantageous populations before treatment and high-risk populations after treatment, enabling optimization of treatment plans and individualized adjustment of follow-up density.

Provided herein are compositions and methods for treating an autoimmune disease or disorder in a subject. In some cases, the compositions and methods use chemical inducers of proximity (CIPs). In some cases, the CIPs include a first moiety having specific binding for a BTB domain-containing protein or a transcriptional repressor expressed in B cell subpopulations and a second moiety having specific binding for a modulator of gene regulation. In some cases, the BTB domain-containing protein comprises BCL6. In some cases, the transcriptional repressor expressed in B cell subpopulations is PRDM1 or IRF4. In some cases the modulator of gene regulation comprises BRD4, CDK9, and / or p300.

The application provides an application of a hair follicle immune homeostasis regulation strategy in treating alopecia diseases. A group of pathological LAMP3 + CCR7 + IL4I1 + DC3 dendritic cell subpopulation. A DC3 dendritic cell subpopulation marker gene Il4i1 Drive conditionally knockout one of the CGRP receptor complexes Calcrl The constructed spontaneous alopecia areata animal model can well simulate the progressive and active hair loss of alopecia areata patients, form stable hair loss patches, and combine with pathological changes such as hair follicle immune cell infiltration, which indicates that the dendritic cell CGRP signal loss is an important inducement for mediating systemic alopecia areata. The application also discloses a pharmaceutical composition or kit for preparing a drug for relieving / treating hair loss, improving hair follicle homeostasis, or enhancing hair growth / black coloration (reducing white hair), which comprises: (a) a JAK kinase inhibitor, and (b) a neuropeptide CGRP or a receptor agonist thereof.

The application relates to the technical field of medical detection, in particular to a kit, a method and a system for predicting bacterial infection. The application provides a kit for predicting bacterial infection, which comprises an antibody combination for specifically recognizing CD64, CD14 and CD14, and the antibodies are respectively labeled with mutually distinguishable fluorescent dyes. Meanwhile, the application provides a method for predicting bacterial infection, which comprises obtaining the counts of neutrophils, lymphocytes and monocytes in a to-be-detected sample, and the fluorescent intensities of CD64 and CD14 on the cell subgroups, and calculating four characteristic parameters IDXNLR, IDXLMR, IDXCD64 and IDXCD14; and then inputting the parameters into a pre-trained infection diagnosis model to output a risk score for judging the infection state (especially bacterial infection). The model significantly improves the accuracy and specificity of distinguishing bacterial infection from non-bacterial infection (such as viral infection or healthy state) by comprehensively integrating immune parameters in multiple dimensions.

Methods are disclosed for adding adapters to fragmented nucleic acids for next generation sequencing, including providing numerical codes based on variable adapter molecular barcode lengths on both sides of the fragmented nucleic acids and identifying reads from the same fragment based on both barcodes. The methods and products allow for the amplification of the fragmented nucleic acids when there is a low yield of isolated fragmented nucleic acids and also for efficient and reliable detection of low-frequency mutations including in subpopulations of cells within a subject.

The application relates to a thymoma epithelial cell subpopulation with neuromuscular characteristics and application, and a thymoma epithelial cell subpopulation with neuromuscular characteristics is obtained through single clone dilution culture screening from a thymoma cell line Thy0517 of a patient with myasthenia gravis (MG) in combination, and a thymoma epithelial cell with neuromuscular characteristics in the cell subpopulation is named as Thymus_NMi; the cell subpopulation has synapse-like structures and neuromuscular adhesion characteristics, and efficiently expresses genes participating in neural cell adhesion and synapse connection, highly integrates neuromuscular double characteristics, and simulates key pathological characteristics of abnormal thymus-induced immune tolerance of MG patients in a molecular phenotype and physiological function, so that a cell model closest to a real clinical state is provided for exploring a myasthenia gravis occurrence mechanism.

The present application relates to the technical field of medical diagnosis, in particular to a flow cytometry T cell subset detection automatic classification method based on machine learning, cell data of multiple blood samples is collected by using a flow cytometer, and a two-dimensional scatter plot is drawn; the cell population in the two-dimensional scatter plot is manually labeled, and the labeling result is mapped to high-dimensional data; the high-dimensional data is standardized to obtain high-dimensional space data; the high-dimensional space data is reduced in dimension based on a uniform manifold approximation and projection algorithm to obtain low-dimensional data; the low-dimensional data is density clustered based on a density-based spatial clustering algorithm, and the classification result is matched with the manual labeling; and the classification result is mapped back to the two-dimensional scatter plot for visual display, which overcomes the problems of strong subjectivity, low efficiency and poor reproducibility in the traditional manual classification method, and significantly improves the accuracy, efficiency and consistency of T cell subset classification.

The application discloses a cell subpopulation automatic annotation method and device, and relates to the technical field of single-cell sequencing. The method comprises the following steps: acquiring single-cell transcriptome data; in a preset target resolution range, an initial clustering result of each resolution, i.e., a first target cluster, is obtained by means of a community discovery algorithm based on modularity optimization; if there is a cluster in the first target clustering result at each resolution that does not meet the specificity requirement, a second target clustering result is obtained by using similarity iterative merging; a representative marker of each cluster in the second target clustering result is determined by using a getSpecificityScore function; and a cell subpopulation annotation result under the resolution is recommended by comparing the specificity score. By means of a multi-scale analysis strategy, the application combines the specificity score and similarity merging cell subpopulation automatic annotation, realizes automatic identification and annotation of novel cell subtypes without relying on prior annotation information, can adapt to diversified data characteristics, and improves the accuracy and robustness of cell subpopulation annotation.

This invention provides a neurodevelopmental drug screening system and its applications based on a set of microglia markers. Specifically, this invention provides a neurodevelopmental supportive microglia subset that expresses markers of neurogenesis and development, as well as energy metabolism. This invention also provides a neurodevelopmental drug screening system and method using changes in the state of this cell subset as the core evaluation indicator. The system of this invention can sensitively and quantitatively reflect the regulatory effect of candidate drugs on the pro-neural maturation function of microglia in developmental brain injury, filling the technical gap in existing drug screening systems that lack "developmental immune-metabolic-synaptic linkage indicators."

The application relates to the technical field of biological medicine, and particularly discloses a monoclonal antibody combined with human CD4 and a preparation method and application thereof. The antibody is obtained by combining single B cell sorting, gene cloning and mammalian cell expression technology after rabbit immunization of a CD4 polypeptide antigen. The antibody has high affinity and good specificity, and can effectively recognize CD4 positive T cells in human peripheral blood. The antibody is verified by ELISA and flow cytometry, and has high titer, low background and a signal-to-noise ratio superior to that of existing mouse-derived antibodies. Further, the amino acid sequences of the light chain and the heavy chain variable region of the antibody are obtained, so that controllability and repeatability of antibody expression are realized. The antibody can be widely applied to scenes such as immunodetection, HIV auxiliary diagnosis, T cell subgroup research and antibody engineering development, and has important scientific research and industrial values.

The present application provides identification of a new visceral adipose tissue Treg cell subpopulation and its application. Specifically, the present application identifies two visceral adipose tissue Treg cell subpopulations: 1) ICOS hi Treg cell subpopulation that specifically highly expresses Icos, Nkg7, Itgb1, Tmsb10, Ms4a4b and Ccl5; and 2) KLRG1 hi Treg cell subpopulation that specifically highly expresses Atf3, Lmna, Klrg1, S100a6 and Cxlc2. Experimentally verified, the new Treg cell subpopulation can improve lipid metabolism, reduce body weight, reduce fat inflammation, thereby effectively improving obesity in vivo.

The application discloses a targeted cytokine composition for immunotherapy and a preparation method thereof. The fusion protein comprises, from N-terminal to C-terminal, an anti-PD-L1 single-domain antibody, a hinge region, a double-mutated human IL-15, a hinge region, an anti-NKG2D single-domain antibody and a double-mutated human IgG1 Fc fragment. In vitro experiments show that the fusion protein can simultaneously bind to double targets, significantly promotes NKG2D-positive CD8-positive T cell proliferation and has weak effect on regulatory T cells, and presents synergistic effect in a tumor-immune cell co-culture system. In vivo pharmacodynamic research shows that in a humanized mouse model, the fusion protein significantly inhibits the growth of colon cancer, the number of CD8-positive T cell and NK cell infiltrations in the tumor is obviously increased, and the effect is better than that of non-targeting and single-targeting controls. The fusion protein provided by the application has long-acting circulation, tumor targeting enrichment and immune cell subpopulation precise regulation functions, and can be used for preparing an antitumor drug.

The invention discloses a single cell 3'end transcriptome variable polyadenylation (APA) analysis method, and belongs to the field of bioinformatics. The analysis process provided by the invention is based on 3'end transcriptome sequencing data of a single cell, combines multiple processes of UMI deduplication, false comparison removal, site merging, internal amplification removal, polyA site removal with few supporting cells, site verification and the like, and can realize efficient detection and quantification of APA at the single cell level. The method is especially suitable for single cell APA analysis of single cell 3'end transcriptome sequencing data, including 3 'end single cell sequencing data after 10x scRNA-Seq cell subset combination, 3' end single cell sequencing data based on a C1 chip, and even full-length transcriptome data.

The present invention provides a chimeric antigen receptor based on the intracellular domain, comprising sequentially linked extracellular domains, transmembrane domains, and intracellular domains, wherein the extracellular domain comprises an antigen recognition region and a hinge region, and the intracellular domain comprises sequentially linked CD3ε intracellular domain cleavage, a co-stimulatory signaling region, and a CD3ζ intracellular domain. Compared to prior art, the provided chimeric antigen receptor exhibits superior therapeutic effects and fewer side effects, as reflected in improved membrane surface CAR molecule levels of CAR-T cells, better immune synapse formation, superior antigen sensitivity, fewer cytokines, better growth capacity, better sustained proliferation capacity, better repeated killing capacity, reduced NK cell activation and cell depletion, and an increase in stem cell-like and memory cell subsets of CAR-T cells. Furthermore, the chimeric antigen receptor can further enhance the therapeutic effect on tumors and, at the same time, reduce the production of inflammatory cytokines associated with the activation of macrophages and monocytes by downregulating its own cytokines, thereby enabling early prevention of cytokine storms.

The application belongs to the technical field of antibody preparation, and particularly relates to an antibody against B lymphocyte antigen CD19, an antibody conjugate and application thereof in immune detection. The amino acid sequences of CDR1-3 on the light chain variable region of the antibody are respectively shown as SEQ ID NO. 3-5, and the amino acid sequences of CDR1-3 on the heavy chain variable region are respectively shown as SEQ ID NO. 8-10. The antibody of the application has high recognition sensitivity and binding affinity to CD19 protein naturally expressed by human and monkey cells, has no cross reaction or weak non-specific binding to non-target proteins on cells, is beneficial to improving the sensitivity, specificity, accuracy and detection precision of detection, provides an antibody tool with good performance, reliability and practicality for the fields of immune cell subpopulation typing and B lymphocyte function identification, and is particularly suitable for the establishment and application of a flow cytometry and multi-color flow cytometry detection system.

This invention discloses a method and application for screening polymorphic genetic markers of disease resistance traits in livestock, relating to the fields of animal molecular genetics and genome breeding technology. This invention uses single-cell transcriptome sequencing to locate key cell subpopulations for disease resistance, and then conducts targeted eQTL analysis on these purified subpopulations. This addresses the shortcomings of relying on mixed samples, which leads to a lack of cell type specificity in markers. Immune cells are grouped using single-cell transcriptome sequencing, and key cell subpopulations in the disease resistance response are screened using pivotal scoring. Cell sorting technology is then used to purify these subpopulations, ensuring that subsequent eQTL analysis is conducted only on these core subpopulations. This process clearly distinguishes the specific cell types involved in the genetic variation, enabling the screened eQTL markers to be directly associated with disease resistance regulatory mechanisms. This achieves a precise correspondence between markers and cell functions, eliminating the interference of mixed samples on marker specificity and giving the markers clear cell type specificity and functional targeting.

This invention belongs to the field of cell preservation technology, specifically disclosing a cell preservation solution. The cell preservation solution of this invention contains a buffer solution, an osmotic pressure regulator, a metal ion chelating agent, and a cell stabilizer. The concentration of the buffer solution in the cell preservation solution is 10-15 mM, the concentration of the osmotic pressure regulator is 0.1-0.3 M, the concentration of the metal ion chelating agent is 2-5 mM, and the concentration of the cell stabilizer is 30-40 mM. The cell preservation solution of this invention not only enables long-term stable preservation of cells under room temperature or refrigeration conditions, but also results in clearer peaks and fewer impurities during subsequent subpopulation analysis such as flow cytometry, leading to better cell subpopulation segmentation. This invention also discloses a method for preparing the cell preservation solution and its application in cell preservation.

The application relates to a lymphocyte subpopulation and cytokine combined detection method, kit and application based on flow cytometry. The method constructs a parallel detection system of cell phenotypes and cytokines on the same flow platform, solves the signal interference problem by using spectrum compatible markers, and realizes synchronous analysis of immune cell subpopulations, immune checkpoints and multiple cytokines in trace blood samples. The scheme avoids system errors and sample waste caused by multi-platform detection, can provide a comprehensive atlas reflecting the synergistic state of cell immune structure and humoral immune function, and improves the accuracy and efficiency of clinical immune evaluation.

This invention provides the application of JAK1 inhibitors in the preparation of drugs for treating adverse reactions related to immune checkpoint blockade. The CD8 inhibitor of this invention... + CTLs in T cells irAE Subgroups are key mediators of adverse reactions associated with multi-organ immunotherapy, specifically overexpressing JAK1 and activating the JAK-STAT signaling pathway, while anti-tumor CD8... + T cell subset 1 (CTL1) shows low JAK1 expression. Inhibitors specifically targeting JAK1 can block CTL expression. irAE‑Ⅱ Inhibiting the JAK-STAT pathway in subpopulations alleviates adverse reactions in multiple organs, including the heart and lungs, without affecting the anti-tumor CD8 inhibitors. + The anti-tumor function of T cell subsets can also enhance the efficacy of immune checkpoint inhibitors.

This invention discloses a rapid method for detecting calcium signals in undifferentiated and differentiated spermatogonia. The method uses a mild enzymatic digestion method to prepare a testicular single-cell suspension. Indo-1 AM, Thy-APC, and c-kit-PE are used for staining and labeling. The cell population to be analyzed is determined based on the difference between the lateral and forward scattering areas of the cells in the testicular cell sample. Then, Thy-APC and c-kit-PE are used to label undifferentiated and differentiated spermatogonia, respectively, to further select specific cell groups. Finally, the fluorescence intensity changes of indo-1 AM in each specific cell subpopulation are detected, thereby obtaining the changes in calcium ion concentration in undifferentiated, undifferentiated-to-differentiated, and differentiated spermatogonia. This method can rapidly analyze the calcium ion concentration in undifferentiated, undifferentiated-to-differentiated, and differentiated spermatogonia. The method is simple and convenient, and provides a foundation for identifying spermatogonial dysfunction caused by calcium signal imbalance.

This invention discloses a novel CD8 + T cells and CD4 + T cell subsets. The CD8... + T cell subsets include: subsets characteristically expressing GZMK and PDCD1; subsets characteristically expressing GZMK, GPR183, IL7R, and ZNF683; subsets characteristically expressing GZMK and CXCL13; subsets characteristically expressing GZMK and ZNF683; and subsets expressing at least one of the following KIR genes: KIR2DL1, KIR2DL3, KIR2DL4, KIR3DL1, KIR3DL2, and KIR3DL3. The CD4+... + T cell subsets include: subsets that characteristically express FOXP3, TNFRSF9, and CTLA4; and subsets that characteristically express FOXP3 and BACH1.

Embodiments described herein generally provide for expanding cells in a cell expansion system. The cells may be grown in a bioreactor, and the cells may be activated by an activator (e.g., a soluble activator complex). Nutrient and gas exchange capabilities of a closed, automated cell expansion system may allow cells to be seeded at reduced cell seeding densities, for example. Parameters of the cell growth environment may be manipulated to load the cells into a particular position in the bioreactor for the efficient exchange of nutrients and gases. System parameters may be adjusted to shear any cell colonies that may form during the expansion phase. Metabolic concentrations may be controlled to improve cell growth and viability. Cell residence in the bioreactor may be controlled. In embodiments, the cells may include T cells. In further embodiments, the cells may include T cell subpopulations, including regulatory T cells (Tregs), helper, naïve, memory, or effector, for example.

The application provides an enhancer for enhancing CAR-T cell expansion and memory phenotype maintenance and an application thereof, relates to the technical field of biological medicine, and adds a small molecule compound IWR-1-endo to the activation of T cells, virus transduction and / or a culture system in a subsequent expansion culture stage, so that the in-vitro expansion efficiency of CAR-T cells can be significantly improved, a larger number of effector cells can be obtained in the same culture time, T cells are effectively inhibited to differentiate into terminal effector cells, the development of the T cells to CAR-Tscm cells and CAR-Tcm subgroups with long-term activity and memory potential is guided and maintained, the product quality is improved, the specific killing capacity on target tumor cells is higher in-vitro, tumors are more effectively removed and long-term control is achieved in an animal model, stronger in-vivo persistence and anti-tumor efficacy are shown. A new strategy for efficiently expanding CAR-T cells in-vitro is provided.

The application discloses a bispecific single-domain antibody (VHH) tandem structure with synergistic effect, a chimeric antigen receptor (CAR) containing the structure, and application of the tandem structure in preparation of anti-tumor immune cells. The tandem structure is sequentially connected by a VHH targeting CD133, a flexible connecting peptide, and a VHH targeting EGFRvIII, and the specific N-terminal to C-terminal sequence (VHH-CD133-linker-VHH-EGFRvIII) is found to be capable of significantly increasing the proportion of the central memory T cell (T_CM) subpopulation in the CAR-T cell and enhancing the penetration ability of the CAR-T cell to solid tumor spheroids. The CAR-T cell containing the tandem structure exhibits better in-vitro and in-vivo synergistic killing activity, persistence and survival benefit than conventional single-target CAR and double-target CAR constructed based on a single-chain antibody (scFv) when treating a heterogeneous tumor expressing CD133 and / or EGFRvIII, in particular, a glioblastoma. The application provides a novel and efficient CAR-T product for overcoming tumor antigen heterogeneity and solid tumor infiltration obstacles.

This invention discloses a method, targets, and applications for the diagnosis and treatment of hemophilic arthritis. The method includes: obtaining cartilage and synovial membrane samples from patients with hemophilic arthritis undergoing total knee arthroplasty; performing single-cell RNA sequencing, single-cell ATAC sequencing, and spatial transcriptomics analysis to determine the main cell types and key cell subpopulations constituting the microenvironment of hemophilic arthritis; identifying the molecular characteristics and differentiation processes of chondrocyte and synovial fibroblast subpopulations; measuring intercellular interactions in the knee joint microenvironment of the samples; and identifying specific regulatory transcription factors in hemophilic arthritis through paired epigenetic analysis to identify diagnostic and therapeutic targets for hemophilic arthritis. This method integrates single-cell multi-omics analysis to analyze the cellular characteristics of synovial and cartilage tissues and their interactions in the epigenetic pathogenic mechanism, identifying potential therapeutic targets.

The present application relates to the application of spatial transcriptome in lung cancer brain metastasis and / or targeted therapy, and in particular to a method for determining a marker epithelial cell subpopulation for lung cancer targeted therapy or lung cancer brain metastasis, which comprises: performing spatial transcriptome sequencing on a to-be-tested sample to obtain gene expression profile data containing spatial position information, taking single-cell transcriptome data of lung cancer tissue as a reference, performing deconvolution analysis on the gene expression profile data containing spatial position information to obtain a cell subpopulation containing spatial position information, circumscribing epithelial cells, performing dimension reduction and cluster analysis to obtain epithelial cell subpopulation types and their proportions, and finding out a marker epithelial cell subpopulation for lung cancer targeted therapy or lung cancer brain metastasis. This method makes full use of spatial position information to analyze cell characteristics and molecules in the development process of lung cancer targeted therapy and brain metastasis, and obtains a marker epithelial cell type and a marker expressed gene, which is of great significance.