41 results about "In vitro stimulation" patented technology

The invention relates to the field of biological medicines, in particular to a tumor antigen peptide and application thereof. The tumor antigen peptide provided by the invention comprises a peptide fragment with an amino acid sequence as shown in SEQ ID No.1. The tumor antigen peptide provided by the invention can be presented by the highest frequency HLA subtype (HLA-A * 11: 01) of Chinese population. In addition, through an in-vitro stimulation mode, it is verified that the tumor antigen peptide can effectively stimulate specific T cells, and a specific TCR element is successfully obtained. Therefore, a direct foundation is laid for further developing a TCR-T therapy, a therapeutic vaccine and other immunotherapies of the targeted human TP53R248Q.

The present application relates to the field of biomedicine, and in particular to a tumor antigen peptide and a use thereof. Provided is a tumor antigen peptide, comprising a peptide segment having an amino acid sequence as shown in SEQ ID NO: 1. The provided tumor antigen peptide can be presented by the HLA subtype (HLA-A*11:01) having a high proportion of Chinese population. Additionally, by means of in vitro stimulation, the tumor antigen peptide is verified to effectively stimulate specific T cells, and a specific TCR element is successfully obtained. The present application lays a direct foundation for further developing TCR-T therapy targeting human TP53 R248Q and immunotherapy such as therapeutic vaccines.

The invention discloses application of ubiquitin specific protease USP4 in treatment of acute lung injury / acute respiratory distress syndrome. The GEO database analysis finds that USP4 is increased in patients with acute respiratory distress syndrome; the expression level of USP4 is increased by stimulating human neutrophils in vitro with LPS, and experiments prove that the USP4 inhibitor Vialinin A can effectively treat acute lung injury and effectively improve the survival rate of mice; acute lung injury can be effectively relieved by using adenovirus to inhibit expression of USP4 in lung; the USP4 defect of the hematopoietic system can effectively relieve acute lung injury. The application of the USP4 serving as the target spot in screening the medicine for treating the acute lung injury or the acute respiratory distress syndrome is provided for the first time, a new target spot is provided for treating the acute lung injury or the acute respiratory distress syndrome, and a new treatment strategy is provided for clinical treatment.

This invention discloses a veterinary aluminum hydroxide adjuvant for foot-and-mouth disease (FMD) vaccines and its preparation method via aluminum alkoxide hydrolysis. The adjuvant is a boehmite phase, and through innovative processing, the crystallinity and morphology are synergistically controlled, achieving a relative crystallinity of 10%-25%. The primary particles are rod-shaped or needle-shaped, with an average aspect ratio of 4:1 to 8:1. The preparation method includes: dissolving an aluminum alkoxide compound, then subjecting it to a controlled hydrolysis reaction at 50-80℃ and pH 4.5-6.5 under normal pressure for 36 hours, followed by aging at 425℃ for 2472 hours. The product is obtained after solid-liquid separation, washing, and drying. The obtained adjuvant exhibits an adsorption rate of over 96% for FMD virus inactivated antigen within 1 hour, and a cumulative antigen dissociation rate of less than 5% after 7 days at 37℃. In vitro stimulation of macrophages to secrete IL-1β and TNF-α levels is more than 1.7 times that of commercially available aluminum hydroxide adjuvants, significantly superior to traditional aluminum hydroxide adjuvants, providing a new material for the preparation of highly effective FMD vaccines.

The invention discloses application of riboflavin in preparation of medicines for treating colorectal cancer, belongs to the field of development of new application of medicines, and particularly relates to application of riboflavin to in-vitro stimulation of primary CD8 + T cells and improvement of the activity of the CD8 + T cells to improve the capacity of killing colorectal cancer cell lines. Meanwhile, the invention also proves that the activity of immune cells in the body of a colorectal cancer patient can be obviously activated through a conventional drinking water preparation and riboflavin in an animal model for the first time, particularly through the activity of CD8 + T cells, so that the capability of killing tumors is improved, and the volume and weight of the tumors are obviously reduced; the riboflavin can be used for preparing the medicine for treating the colorectal cancer.

The invention discloses a Brucella bacterium capable of generating a lipoid A vaccine adjuvant and an application of the Brucella bacterium. According to the invention, recombinant plasmids for expressing BacA and LpxL genes are constructed, and are transformed into Brucella through electric shock, so that a double-gene knockout strain is successfully obtained. By knocking out BacA, an ultra-long fatty acid chain of lipoid A is shortened, an organism is induced to generate an inflammatory reaction, and by knocking out LpxL, the inflammatory reaction is reduced, so that the effect of the immunologic adjuvant is achieved. LPS is extracted for in-vitro stimulation analysis, and it is found that the inflammatory response of double-gene deletion is reduced compared with that of single-gene deletion. Animal experiments show that double-gene deletion can induce an organism to generate good immune response and induce relatively high cellular immunity and humoral immunity, and a new thought is provided for research and development of gram-negative bacterium inactivated vaccines.

The invention relates to the technical field of deep brain electrical stimulators, and discloses a deep brain electrical stimulator for treating heart and cerebral vessels, which comprises an in-vitro stimulation generator and an extension wire, the device further comprises a head fixing assembly and a control assembly, the left side of the in-vitro stimulation generator is movably connected with the head fixing assembly, the control assembly is installed on the upper portion in the in-vitro stimulation generator, the output end of the control assembly is connected with an extension wire, the head fixing assembly comprises a spring belt, and the spring belt is arranged on the left side of the in-vitro stimulation generator. Fixing plates are connected between the spring belts in a central symmetry mode, mounting boxes are movably connected to the opposite sides of the fixing plates, and rubber plates are movably connected to the opposite sides of the mounting boxes; through the arrangement of the spring belt and the mounting box, the electrode can be more tightly attached to the skull of a patient, so that the stability of the electrode is improved, the situation that the treatment effect is affected by displacement or falling of the electrode is avoided, meanwhile, the spring belt can be adjusted according to the shape and size of the head of the patient, and a more personalized fixing mode is provided.

The invention discloses a TIL cell culture medium and a TIL cell culture method, an activation and proliferation culture medium of TIL cells comprises the following components: a T cell serum-free culture medium, IL-2, platelet lysate and gentamicin, a CD3 monoclonal antibody, a CD28 monoclonal antibody and phytolectin PHA-P are added for cell activation, and then the activation and proliferation culture medium is used for activation and proliferation culture. Wherein CD3 and CD28 antibodies are used in combination to stimulate TIL cells and simulate the double-signal effect of in-vivo TIL cell activation, meanwhile, PHA-P is utilized, by means of the characteristics of PHA-P, small gonorrhea cells are stimulated in vitro to be converted into gonorrhea cells with immunocompetence, division of the gonorrhea cells is promoted, and the proliferation capacity of the TIL cells is improved through the synergistic effect of PHA-P and IL-2; platelet lysate is used for replacing plasma of a patient and is used as an efficient culture supplement for supporting TIL cell growth; finally, more TIL cells are obtained through the technology, and the number of immune cells required by clinical effective treatment can be sufficiently met.

Disclosed herein are methods of stimulating and expanding polyclonal gamma delta T cells (GDTCs) in vitro. More specifically, anti-gamma delta T cell receptor (GDTCR) antibody is used in combination with anti-CD28 antibody and one or more of IL-2, IL-7, and IL-15 to efficiently expand GDTCs in vitro.

Some embodiments of the methods and compositions provided herein relate to the use of hapten labeled cells to stimulate chimeric antigen receptor (CAR) T cells. In some embodiments, CAR T cells can include a CAR that specifically binds to a hapten. Some embodiments relate to the in vivo or in vitro stimulation CAR T cells by hapten labeled cells.

To provide peptides, proteins, nucleic acids and cells for use in immunotherapeutic methods.SOLUTION: In particular, the present invention relates to the immunotherapy of cancer. The present invention furthermore relates to tumor-associated T-cell peptide epitopes, alone or in combination with other tumor-associated peptides, which can serve, for example, as active pharmaceutical ingredients of vaccine compositions that stimulate anti-tumor immune responses, or stimulate T cells ex vivo and transfer into patients. Peptides bound to molecules of the major histocompatibility complex (MHC), or peptides as such, may also be targets of antibodies, soluble T-cell receptors, and other binding molecules.SELECTED DRAWING: None

The invention discloses a tumor neoantigen polypeptide aiming at BRD4: NUTM1 gene fusion mutation and an application of the tumor neoantigen polypeptide in NUT cancer treatment. The tumor neoantigen polypeptide comprises: (1) a tumor neoantigen polypeptide with an amino acid sequence represented by SEQ ID No.2; and (2) polypeptides with the same or similar functions obtained by substitution and / or deletion and / or addition of at least one amino acid in the amino acid sequence. The antigen peptide disclosed by the invention can stimulate and activate T cells specifically aiming at BRD4:: NUTM1 gene fusion mutation of a human body in vitro and perform mass amplification for adoptive reinfusion treatment of a patient; the polypeptide has obvious immunogenicity, and the killing ability of T cells on NUT cancer cells with BRD4: NUTM1 gene fusion mutation is improved; meanwhile, the BRD4: NUTM1 fusion mutant tumor antibody can be synthesized on a large scale and is used for subsequent standardized and individualized immunotherapy of BRD4:: NUTM1 fusion mutant tumor patients.

The invention discloses a method for in-vitro broad-spectrum stimulation of immune checkpoint up-regulation and application, the method comprises the following steps: placing a Jurkat cell line in an RPMI-1640 culture medium containing 10% of fetal calf serum and 1% of penicillin-streptomycin, carrying out in-vitro stimulation for 24-72 hours by adopting PHA with a concentration of 0.1-5 [mu] g / mL under the conditions of 37 DEG C and 5% of CO2, activating NF-kB and / or MAPK signal channels, and carrying out in-vitro broad-spectrum stimulation of immune checkpoint up-regulation. The broad-spectrum up-regulation of co-stimulatory immune checkpoints of CD28, CD40L, ICOS and the like and co-inhibitory immune checkpoints of PD-1, LAG-3, CTLA-4 and the like is realized, and the high expression state can be maintained for at least 72 hours after the PHA is removed. The invention also discloses the in-vitro broad-spectrum immune checkpoint positive T cell model constructed by the method, and the up-regulation of immune checkpoints in the model is accompanied by the increase of dose-dependent secretion of various cytokines. The model can replace a traditional model, is applied to toxicology and pharmacology research of immune checkpoint antagonists and combined synergistic effect and immune checkpoint regulation and control mechanism research, is simple and convenient to operate and high in repeatability, and provides an efficient tool for research and development of related drugs.

The present application relates to a multi-channel extracorporeal stimulator and a control method thereof. The multi-channel extracorporeal stimulator comprises a power module, a controller module, a stimulation module and a program control module. The power module is electrically connected with the controller module and the stimulation module respectively, and supplies power to the controller module and the stimulation module respectively. The program control module is in communication connection with the controller module, and sends preset program control parameter information to the controller module. The controller module is electrically connected with the stimulation module, and generates a power control signal and a stimulation program signal according to the received preset program control parameter information, sends the power control signal to the power module, and sends the stimulation program signal to the stimulation module. The stimulation module generates stimulation waveforms of multiple channels according to the received stimulation program signal. The extracorporeal stimulator and the method can realize more stimulation modes and wider stimulation current amplitude by supporting more stimulation channels, so as to set multiple treatment schemes and achieve the best treatment effect.

The present disclosure provides compositions and methods for stimulating, enriching, expanding, and / or activating engineered cells that express a recombinant receptor, e.g., a chimeric antigen receptor. In some embodiments, the provided methods include ex vivo or in vitro stimulation, enrichment, expansion, and / or activation of cells by incubation with a particle, e.g., a bead particle, attached to a binding molecule, such as a polypeptide antigen or anti-idiotype antibody, that recognizes or binds to the recombinant receptor. Also provided herein are methods for transfecting or transducing cells that have not been previously incubated with an activating or stimulating agent, such as have not been incubated with anti-CD3 / anti-CD28 antibodies and / or one or more recombinant cytokines, by transducing or transfecting the cells in the presence of the particles with attached binding molecules. In some embodiments, the provided compositions can be used in methods to prepare cells, e.g., genetically engineered T cells, for of adoptive immunotherapy.

To provide peptides, proteins, nucleic acids and cells for use in cancer immunotherapeutic methods.SOLUTION: The present invention relates to tumor-associated T-cell peptide epitopes, alone or in combination with other tumor-associated peptides, which can serve as, for example, active pharmaceutical ingredients of vaccine compositions that stimulate anti-tumor immune responses, or to stimulate T cells ex vivo and transfer into patients. Peptides bound to molecules of the major histocompatibility complex (MHC), or peptides as such, can also be targets of antibodies, soluble T-cell receptors, and other binding molecules.SELECTED DRAWING: None

The present invention discloses a mesenchymal stem cell derived from a definitive endoderm cell, a differentiation method and an application thereof, and relates to the technical field of mesenchymal stem cells. The differentiation method comprises: inducing human pluripotent stem cells to differentiate into mesendoderm cells by culturing in a first differentiation medium containing a WNT pathway activator and Activin A for 1-2 days; inducing the formation of definitive endoderm cells (DE) by culturing in a second differentiation medium containing no WNT pathway activator and Activin A for 2-8 days; inducing the acquisition of midgut, hindgut and foregut cell fate by culturing in a MSC generation medium for 2-4 days; inducing the generation of a cell population containing mesenchymal stem cells by culturing in a MSC generation medium for 8-10 days; performing digestion and passage 2-3 times to obtain mesenchymal stem cells (DE-MSCs) derived from definitive endoderm cells. It can secrete anti-inflammatory factors under in vitro stimulation and has the potential to treat colitis.

Provided herein are methods of producing a plurality of T cells which has been ex vivo stimulated and expanded to enrich for antigen-specific T cells. This plurality of T cells can be used to provide highly specific and efficacious T cell therapies for treatment of cancers and other conditions.

Disclosed are methods of obtaining a cell population enriched for T cells having antigenic specificity for a cancer-specific mutation using in vitro stimulation of memory T cells. Also disclosed are related methods of isolating a T cell receptor (TCR), populations of cells, TCRs or antigen-binding portions thereof, pharmaceutical compositions, and methods of treating or preventing cancer.

The present invention relates to the prevention and treatment of disease like cancer. The inventors have previously characterized MELOE-1 antigen as an IRES dependent, melanoma specific translation product from a lncRNA mainly transcribed in the melanocytic lineage. MELOE-1 contains numerous class II epitopes and one HLA-A*0201-restricted CD8 epitope eliciting a frequent repertoire of high avidity T cells. They designed various synthetic long peptide (SLPs) comprising a CD4 epitope coupled to the CD8 epitope by a serie of linkers of 4 to 6 aa and studied the efficacy of T cell clone activation by SLP-loaded DC in vitro. Particularly, they evaluated the ability of a few selected SLPs to stimulate specific T cells proliferation of PBL from healthy donors in vitro and finally, they explored the vaccination potential of their best SLP candidate in vivo in an HLA*A0201 / HLA-DRB0101 transgenic mouse. Thus, the present invention relates a SLP comprising a CD4 class II peptide linked to a CD8 class I peptide by a specific linker and its use in the treatment of disease like cancers.

The present invention discloses a humanized anti-human γδTCR monoclonal antibody and its encoding gene, preparation method and application, which belongs to the humanized monoclonal antibody preparation technology field in the biotechnology field. The heavy chain variable region of the humanized anti-human γδTCR monoclonal antibody provided by the present invention comprises the amino acid sequence shown in SEQ ID NO:4 or SEQ ID NO:5, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:7. The humanized anti-human γδTCR monoclonal antibody can stimulate and amplify peripheral blood γδT cells in vitro, and the obtained γδT cells can secrete a variety of cytokines and can effectively kill a variety of tumor cells.

The present invention relates to peptides, proteins, nucleic acids and cells for use in immunotherapeutic methods. In particular, the present invention relates to the immunotherapy of cancer. The present invention also relates to tumor-associated T cell peptide epitopes, alone or in combination with other tumor-associated peptides (active pharmaceutical ingredients of vaccine complexes that stimulate anti-tumor immune responses or stimulate T cells ex vivo and transfer into patients). Peptides bound to molecules of the major histocompatibility complex (MHC), or peptides as such, may also be targets of antibodies, soluble T-cell receptors, and other binding molecules.

This invention discloses an electromagnetic modulation method for improving the formation of glial scars around implanted electrodes. The method selects at least two electrode channels in an implanted brain-computer interface electrode array as stimulation channels and connects them to an external stimulation driving circuit. By outputting a charge-balanced low-frequency pulse stimulation current, it implements staged modulation of the local tissue around the electrodes at different stages after implantation. Simultaneously, it continuously monitors the electrode-tissue interface impedance and / or the signal-to-noise ratio of neural signals, and performs closed-loop switching between different stimulation parameters based on the monitoring results. This method is drug-free and can utilize bipolar or tripolar current-guided stimulation configurations to regulate the spatial distribution of the local electric field. Combined with the accompanying near-field electromagnetic effect, it provides electromagnetic parameter-dependent long-term modulation of the inflammatory response of microglia and astrocytes at the implantation interface, thereby improving the long-term electrical stability of the implantation interface and extending the lifespan of the implanted electrodes.

Disclosed are methods of obtaining a cell population enriched for T cells having antigenic specificity for a cancer-specific mutation using in vitro stimulation of memory T cells. Also disclosed are related methods of isolating a T cell receptor (TCR), populations of cells, TCRs or antigen-binding portions thereof, pharmaceutical compositions, and methods of treating or preventing cancer.

The application belongs to the field of medical biotechnology, and particularly relates to a method for in-vitro ADP stimulation determination of platelet activation and application. The method provided by the application comprises: S1, adding buffer solution, ADP solution and anticoagulant whole blood sample into a homologous control tube and a to-be-detected tube respectively; S2, placing the homologous control tube and the to-be-detected tube at 18-28 DEG C for 5-30 min for stimulation; the final concentration of the ADP solution in the step S1 is 5-160 micromol / L; and the addition amount of the anticoagulant whole blood sample is 5-10 microliters. The method provided by the application is more direct and simple to operate, and only needs to be completed at normal temperature. After the stimulation is completed, the antibody reagent is directly added without the need of taking a flow tube for secondary dispensing and adding the antibody reagent. In addition, the sample amount of the method provided by the application is significantly reduced, the burden of patients is reduced, the ADP amount is greatly reduced, the reagent amount is saved, the collection time is significantly shortened, and the detection efficiency is greatly improved.

The present invention relates to peptides, proteins, nucleic acids and cells for use in immunotherapeutic methods. In particular, the present invention relates to the immunotherapy of cancer. The present invention furthermore relates to tumor-associated T-cell peptide epitopes, alone or in combination with other tumor-associated peptides that can for example serve as active pharmaceutical ingredients of vaccine compositions that stimulate anti-tumor immune responses, or to stimulate T cells ex vivo and transfer into patients. Peptides bound to molecules of the major histocompatibility complex (MHC), or peptides as such, can also be targets of antibodies, soluble T-cell receptors, and other binding molecules.

To provide novel reagents to improve the in vitro and in vivo expansion of CAR T cells for ACT.SOLUTION: The present application is directed to bispecific polypeptides comprising a first binding domain binding an antigen on an antigen presenting cell (ARC) and a second binding domain binding an antigen on an immune cell expressing a chimeric antigen receptor (CAR). Nucleic acids, vectors and host cells used in producing the polypeptide of the invention are also disclosed. Compositions comprising the bispecific polypeptides and methods of treating cancer and stimulating activation and expansion of immune cells in vivo and in vitro are also disclosed.SELECTED DRAWING: None

The invention discloses an application of an anti-lymphocyte function associated antigen 1 (LFA-1) neutralizing antibody in a medicine for relieving neuroinflammation of Parkinson's disease, and belongs to the technical field of biological medicines. The invention further discloses a preparation method of the anti-lymphocyte function associated antigen 1 (LFA-1) neutralizing antibody and an application of the anti-lymphocyte function associated antigen 1 (LFA-1) neutralizing antibody in the medicine for relieving the neuroinflammation of the Parkinson's disease. The method comprises the following steps: co-culturing Teff cells stimulated and activated in vitro or Teff cells activated after being treated by using an LFA-1 neutralizing antibody with MN9D cells of an MPP + untreated group and an MPP + treated group for 48 hours, and observing the influence of the Teff cells on the activity of the MN9D cells, the number of apoptotic and necrotic cells, LDH release and apoptotic protein expression conditions. Results show that after pretreatment with the LFA-1 neutralizing antibody, MN9D cell viability decline, apoptosis and necrotic cell number increase, LDH release increase and apoptosis protein expression up-regulation caused by Teff cells are all relieved (Plt; 0.05) of the method.

This invention discloses the application of ubiquitin-specific protease USP4 in the treatment of acute lung injury / acute respiratory distress syndrome (ARDS). Analysis of the GEO database revealed elevated USP4 levels in ARDS patients; in vitro stimulation of human neutrophils with LPS showed increased USP4 expression levels; experiments confirmed that the USP4 inhibitor Vialinin A effectively treats ARDS and significantly improves mouse survival; adenovirus inhibition of USP4 expression in the lungs effectively alleviates ARDS; and USP4 deficiency in the hematopoietic system effectively alleviates ARDS. This invention is the first to propose the application of USP4 as a target in screening drugs for the treatment of ARDS, providing a new target and a novel treatment strategy for clinical practice.

The invention discloses a peripheral blood sample stabilizing solution for inhibiting platelet activation through a multi-target synergistic effect and also discloses a composite anticoagulant formula containing the stabilizing solution. The composite anticoagulant formula comprises three parts, namely a stabilizing solution, a platelet antibody detection reagent and a fixing agent. The platelet can be protected or platelet activation can be reversibly inhibited, on one hand, the detection time of the platelet resting state can be prolonged, on the other hand, the activation, agglutination and adhesion functions of the platelet are reserved, and convenient conditions are provided for research and detection of in-vitro stimulation of the platelet.