64 results about "Mouse Spleen" patented technology

The invention provides a monoclonal antibody capable of recognizing a recombinant EGF-CRM197 vaccine and a preparation method thereof, on the basis of researching, developing and preparing the recombinant EGF-CRM197 vaccine, after the recombinant EGF-CRM197 vaccine is further immunized and inoculated to a mouse, mouse spleen cells and mouse myeloma cells are fused, and the monoclonal antibody capable of recognizing the recombinant EGF-CRM197 vaccine is obtained. And screening a hybridoma cell strain which generates a specific monoclonal antibody for recognizing the recombinant EGF-CRM197 vaccine, and sequencing to obtain an antibody sequence. After the hybridoma cells are cultured in a mouse body, the monoclonal antibody for recognizing the recombinant EGF-CRM197 vaccine can be prepared in vitro through methods of chromatographic purification and the like. The method can be used for detecting the release titer of the recombinant EGF-CRM197 vaccine and evaluating the immune efficacy after the recombinant EGF-CRM197 vaccine is immunized.

The invention discloses preparation of a human CD56 immune phage display antibody library, which comprises the following steps: S1, animal immunization: expressing recombinant hCD56 protein in escherichia coli, then immunizing a Balb / c mouse, and detecting immune titer; s2, splenocyte preparation: extracting RNA (Ribonucleic Acid) of immune mouse spleen cells, and performing reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid); s3, preparation of an initial bacteriophage display library: amplifying a heavy chain variable region and a light chain variable region by using a PCR technology, and then assembling to form scFv; constructing scFv phagocytids, electrically transforming the scFv phagocytids into TG1 competent cells, and constructing a bacterial library; s4, constructing a phage single-chain library of the hCD56: infecting the bacterial library by using a helper phage M13K07, and constructing a phage single-chain antibody library; and S5, elutriation and screening of a phage library: specifically enriching a phage single-chain antibody library, screening positive clones from the phage single-chain antibody library, and determining a sequence. According to the invention, the human hCD56 antibody scFv sequence fragment can be screened out, and the research and development period is shortened.

The invention belongs to the technical field of biological medicine and molecular biology, and particularly relates to a single-chain antibody targeting NRP1, a CAR-T cell and application of the single-chain antibody and the CAR-T cell in fibrosis treatment. Four types of high-affinity single-chain antibodies targeting mouse NRP1 antigens are obtained by immunizing mice, the single-chain antibodies are applied to second-generation CAR, T cells of mouse spleen sources are infected after virus packaging, and CAR-T cells are constructed. The CAR-T cells of the targeted NRP1 can be used for effectively killing NRP1 positive cells. In-vivo experiments show that the CAR-T cells can inhibit the growth of NRP1 positive tumor cells. The CAR-T cell can improve the mouse liver fibrosis condition, collagen in the mouse liver is reduced through treatment, and the molecular level of fibrosis markers SMA and NRP1 is reduced, it is indicated that the CAR-T cell recognizes and eliminates activated HSCs through targeting NRP1, and the fibrosis degree is reduced.

The invention discloses a dendrobium huoshanense polysaccharide nanoemulsion immunologic adjuvant and a preparation method thereof. The dendrobium huoshanense polysaccharide nanoemulsion immunologic adjuvant is prepared from the following raw materials in percentage by mass: 0.1%-2.0% of dendrobium huoshanense polysaccharide, 20%-30% of a surfactant, 5%-10% of a cosurfactant, 5%-10% of an oil phase and the balance of deionized water. The preparation method of the nano-emulsion is simple and easy to implement, equipment is easy to control, the cost is low, and the obtained nano-emulsion is small in droplet particle size, high in stability and capable of improving the spleen lymphocyte proliferation activity of immunocompromised mice. The nanoemulsion can significantly improve the immunomodulatory activity of dendrobium huoshanense polysaccharide, and has the potential of developing immunopotentiators, thereby promoting the development and utilization of traditional Chinese medicine resources and active components thereof.

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.

This invention discloses an antibody that specifically recognizes Toxoplasma gondii surface antigens and its applications. The invention utilizes hybridoma technology to establish a hybridoma cell line by fusing Balb / C mouse spleen cells with SP2 / 0 cells. Monoclonal antibodies were prepared from these cells using in vitro culture and in vivo ascites induction methods, yielding the monoclonal antibody SH12-S7. This invention provides clinical and research institutions with an antibody that specifically recognizes Toxoplasma gondii surface antigens. It can be used for ELISA assays to detect clinical samples, for immunohistochemical assays to diagnose pathological specimens, and provides a convenient research tool for toxoplasmosis research departments. It has broad application value in the diagnosis and detection of definitive host infections and provides technical support for epidemiological investigations and evaluation of control effects of Toxoplasma gondii.

The present invention provides the preparation, epitope identification and application of a PRRSV non-structural protein 12 antibody. By constructing a eukaryotic expression vector pCDNA3.4-Nsp12-strep, transiently transfecting CHO cells, and purifying the PRRSV Nsp12 recombinant protein using the downstream strep tag, the purified Nsp12 is used as an antigen to immunize Balb / C mice. The spleen cells of the mice are isolated, and single specific B cells are separated by the FACS method. The antibody variable region sequences of the single B cells are obtained by RT-PCR. Recombinant expression plasmids of the heavy chain and light chain are constructed respectively, transfected into CHO suspension cells, expressed, and 2 complete anti-PRRSV Nsp12 monoclonal antibodies 1N14 and 2S18 are obtained by Protein G purification. The application scopes of the 1N14 and 2S18 antibodies and the specific antigen epitope recognized by the 1N14 antibody are further determined, and its sequence is 104 YEFTGNGEDW 113 。

The invention discloses an application of a natural product WithangatinA in preparation of erythroleukemia induced differentiation treatment drugs, the natural product WithangatinA promotes HEL and K562 cells to be transformed from malignant proliferative cells to normal megakaryocytes by inducing the cell particle size to become larger and significantly increasing multinuclear megakaryocytes and promoting expression of surface labeled molecules differentiated from megakaryocytes at the same time, and the erythroleukemia induced differentiation treatment drugs are obtained. The effect of resisting erythroleukemia is achieved. After the natural product Withangulin A is adopted for treatment, the size and the weight of the spleen of a leukemia mouse can be remarkably reduced, the hematocrit and the platelet number of the mouse are remarkably increased, in addition, treatment of the natural product Withangulin A has no obvious influence on the weight of the mouse, and the safety is high. In-vivo research shows that the medicine can induce megakaryocyte differentiation of erythroleukemia in a mouse body, the effect of resisting erythroleukemia is achieved, and a research basis is provided for development of differentiation treatment medicines of erythroleukemia.

The present invention provides a monoclonal antibody that recognizes a recombinant EGF-CRM197 vaccine and a preparation method thereof. Based on the research and development and preparation of a recombinant EGF-CRM197 vaccine, the present invention further immunizes mice with the recombinant EGF-CRM197 vaccine, fuses the mouse spleen cells with mouse myeloma cells, screens and produces a hybridoma cell line of specific monoclonal antibodies that recognize the recombinant EGF-CRM197 vaccine, and obtains the antibody sequence after sequencing. After the hybridoma cells are cultured in mice, monoclonal antibodies that recognize the recombinant EGF-CRM197 vaccine can be prepared in vitro by methods such as chromatography purification. The present invention can be used for the release potency detection of the recombinant EGF-CRM197 vaccine and for evaluating the immune efficacy after immunization with the recombinant EGF-CRM197 vaccine.

To conduct detection of Salmonella, serotype transversely and specifically relative to other strains.SOLUTION: A heat-killed bacterial cell of Salmonella and antigen liquid containing an adjuvant are inoculated in a BALB / c mouse. After the inoculation, splenocytes or lymph node cells of the mouse and P3U1 are fused to prepare a hybridoma, and therefrom, a hybridoma for producing a monoclonal IgG antibody showing Salmonella specificity is selected, whereby three hybridomas for producing superior Salmonella specific antibodies are successfully acquired. Next, these monoclonal antibodies are used to detect Salmonella of a plurality serotypes and other strains according to sandwich ELISA, thereby obtaining a result in which Salmonella is detected serotype transversely, whereas other strains are not detected.SELECTED DRAWING: Figure 1

The invention discloses a method for purifying a toxoplasma gondii recombinant antigen, which comprises the following steps: selecting a full-length sequence of a toxoplasma gondii surface antigen SRS3, connecting the full-length sequence with a carrier through enzyme digestion to form plasmids, performing mass expression in escherichia coli, sequentially performing ultrasonication and centrifugation to collect cell supernatant, and performing deep filtration; the specific affinity chromatography column is coupled with an IgG subtype antibody obtained by hybridizing splenocytes and myeloma cells of a BALB / c mouse infected with toxoplasma gondii through a hybridoma technology. And through affinity chromatography purification, the toxoplasma gondii antigen protein with relatively high purity is finally obtained. According to the method, the toxoplasma gondii recombinant antigen can be stably and effectively purified from the cell supernatant. The method provided by the invention has the advantages of high extraction purity, simple process, capability of realizing one-step purification, stable antibody and the like, and has important significance for realizing purification of the toxoplasma gondii recombinant antigen.

The invention belongs to the technical field of medicines, and particularly relates to a novel tuberculosis subunit vaccine as well as a preparation method and application thereof, and the preparation method comprises the following steps: S1, culturing a mycobacterium bovis attenuated strain; s2, extracting the cell wall of the attenuated strain of mycobacterium bovis; s3, identifying cell walls of the attenuated strains of the mycobacterium bovis; s4, preparing a cell wall and manganese adjuvant subunit vaccine; s5, subunit vaccine intranasal immunization of mice; and S6, performing a mouse challenge protection experiment. Two novel tuberculosis subunit vaccines named as CW-MnJ (alpha) and CW-MnJ (beta) are prepared by extracting cell wall components of a mycobacterium bovis attenuated strain and using the cell wall components as antigens and combining the cell wall components with manganese adjuvants (MnJ (alpha) and MnJ (beta)). A mouse immune protection experiment proves that the lung and spleen bacterium load can be remarkably reduced by the mouse intranasal immunization CW-MnJ (alpha) and CW-MnJ (beta), and the spleen bacterium load of the mouse immunized with the CW-MnJ (alpha) is remarkably reduced compared with a positive control group (BCG immunization group), so that the CW-MnJ (alpha) and CW-MnJ (beta) subunit vaccines prepared by the invention have very good immune protection effects.

The invention discloses a 15-antibody combination for accurately detecting a mouse tissue immune cell subset and application of the 15-antibody combination. Aiming at the problems of large fluctuation and instability of detection results caused by adjustment of voltage and fluorescence compensation during detection by a flow cytometer and different processing modes during result analysis, the antibody combination disclosed by the invention takes monoclonal antibodies CD45, CD3, CD4, CD8, CD19, B220, Ly-6C, Ly-6G, MHC-II, CD11c, CD49b, CD62L, CD44 +, PD-1 and PD-L1 as effective components, and a stable and standard detection method for mouse tissue immune cell subsets is established. The method can analyze the proportion, number and distribution of various immune cells (T cells, B cells, NK cells, dendritic cells, mononuclear cells and granulocytes) in spleen, lung, liver, brain or kidney tissues of mice, can reflect the immune state of the mice and evaluate the immunosuppression state, and can be applied to the fields of disease models, drug evaluation, vaccine development and the like.

The present invention belongs to the technical fields of biomedicine and molecular biology, and particularly relates to a single-chain antibody targeting NRP1 and its application in CAR-T immunotherapy for fibrosis. In the present invention, four single-chain antibodies with high affinity targeting mouse NRP1 antigen are obtained by immunizing mice, and are applied to the second-generation CAR. After viral packaging, they are used to infect T cells derived from mouse spleens to construct CAR-T cells. CAR-T cells targeting NRP1 can effectively kill NRP1-positive cells. In vivo experiments show that CAR-T cells can inhibit the growth of NRP1-positive tumor cells. CAR-T cells can improve the condition of liver fibrosis in mice. After treatment, the collagen in the livers of mice decreases, and the molecular levels of fibrosis markers SMA and NRP1 decrease, indicating that CAR-T cells recognize and eliminate activated HSCs by targeting NRP1, reducing the degree of fibrosis.

The application provides application of psoralen in activating NK cells. The psoralen provided by the application is a coumarin component extracted from traditional Chinese medicine psoralea; through in-vitro cell experiment verification, the psoralen can activate NK-92 to secrete IFN-gamma, Granzyme B and Perforion, and activate surface protein NKG2D of NK-92; meanwhile, the psoralen also has the same effect on a cell group containing NK cells; through in-vivo experiment verification, the psoralen helps to improve the total amount of NK cells in the mouse spleen, inhibit tumor growth, and improve the expression amount of IFN-gamma, Granzyme B, Perforion, NKG2D, NKP46 and CD107a in a tumor infiltration environment. In summary, the small-molecule compound psoralen has good effect in activating NK cells.

The invention relates to the technical field of vaccines, in particular to an adjuvant of a pasteurella inactivated vaccine and application of the adjuvant. According to the specific technical scheme, the adjuvant of the pasteurella inactivated vaccine comprises radix tetrastigme polysaccharide. According to the invention, the THP adjuvant is added into the Pm inactivated vaccine to immunize mice; the Pm inactivated vaccine can effectively promote humoral immune response of the Pm inactivated vaccine, promote secretion of serum Th1 type, Th2 type and Th17 type cell factors after mice are immunized with the Pm inactivated vaccine, enhance cellular immune response of the mice induced by the Pm inactivated vaccine, increase the proportion of T lymphocyte subclasses in splenic lymphocytes of the mice inoculated with the Pm inactivated vaccine, and effectively avoid infection of Pm to the mice.

The invention discloses a partridge extraction isolate and application thereof. According to the present invention, the chemical components of the partridge are researched, such that eight compounds are separated, and the eight compounds are respectively identified as 2-(1H-indol-3-yl) ethyl-tricosanoate (2-(1H-indol-3-yl) ethyl-tricosate) (1), lariciresinol (2), pinoresin (3), syringesin (4), dioscin (5), coniferyl aldehyde (6), syringaldehyde (7) and sinapinalde (8); the compounds are separated from partridge for the first time, the compound (1) is a new compound, the immunomodulatory effect of the compound (1) on mouse splenic lymphocytes is measured, and the result shows that the compound (1) can remarkably induce secretion of TNF-alpha and IL-6, and shows that the compound has immune activation potential equivalent to that of a classical TLR agonist.

The invention provides an anti-medetomidine monoclonal antibody as well as application and a product thereof, and relates to the technical field of biology. Medetomidine is used for immunizing a Balb / c mouse, splenocytes of the mouse are fused with myeloma cells, hybridoma cells with high specificity are obtained through specific high-throughput screening, a large amount of mouse ascites is obtained through culture and re-immunization, and the mouse ascites with high specificity is obtained. And the medetomidine monoclonal antibody with high purity, high sensitivity and high specificity is obtained through multi-step separation and purification, so that a required raw material is provided for developing an immune test strip for detecting medetomidine. The anti-medetomidine monoclonal antibody disclosed by the invention can be used for immunological detection such as immunoblotting and immunofluorescence, and the obtained antibody is proved to have good specific binding capacity.

The present application belongs to the field of biomedical technology, and more particularly relates to the application of Periplaneta americana polypeptide HFDT1 as an immune adjuvant in the preparation of a brucellosis vaccine. The present application uses Brucella OMP10, BP26 and L7 / L12 proteins in combination with HFDT1 to induce mice to produce higher titers of anti-OMP10, BP26 and L7 / L12 protein specific IgG antibodies, and the titers increase with the increase in the number of immunizations, while promoting the body to produce mixed Th1 / Th2 type cellular immune responses and stimulating the proliferation and differentiation of mouse spleen T and B cells, and also facilitating mass production. The Periplaneta americana polypeptide HFDT1 described in the present application is obtained by convenient mass synthesis, and can be used as a novel small molecule immune adjuvant to enhance specific immune responses of the body.

The invention provides an anti-trihexylol monoclonal antibody as well as application and a product thereof, and relates to the technical field of biology. The method comprises the following steps: immunizing a Balb / c mouse by using trihexylamine, fusing splenocytes of the mouse with myeloma cells, carrying out specific high-throughput screening to obtain hybridoma cells with high specificity, and carrying out culture and re-immunization to obtain a large amount of mouse ascites. And the monoclonal antibody resisting the trihexylamine with high purity, high sensitivity and high specificity is obtained through multi-step separation and purification, so that a required raw material is provided for developing an immune test strip for detecting the trihexylamine. Through verification, the monoclonal antibody for resisting the trihexylamine disclosed by the invention has good specific binding capacity.

The invention discloses application of sitagliptin in preparation of a medicine for treating Graves eye diseases, and belongs to the field of biological medicine. The invention discloses the application of sitagliptin in preparation of the medicine for treating Graves eye diseases for the first time, and sitagliptin can improve expression of CD4 + CTL cytotoxic functional molecules and improve eye symptoms, extraocular muscle thickening and fibrosis, orbit fat neogenesis and orbit T lymphocyte infiltration of mice; the mouse spleen CD4 + CTL cell proportion is improved; in addition, the thyroid hormone level, thyroid hyperplasia and hypertrophy and lymphocyte infiltration of mice can be improved. Sitagliptin is a clinically common hypoglycemic drug, and is perfect in safety database, small in side effect and easy to popularize clinically. Therefore, the experimental result of the invention fully supports the new application of sitagliptin in treating Graves eye diseases, and is a new direction worthy of exploration for treating Graves eye diseases due to strong clinical operability.

The invention relates to an application of a traditional Chinese medicine composition in preparing a medicine for atopic dermatitis, which is characterized in that the traditional Chinese medicine composition is prepared from astragalus membranaceus, dried ginger, salted morinda officinalis, cortex dictamni, calcined oyster shell, schisandra chinensis and divaricate saposhnikovia root. The composition provided by the invention can obviously inhibit increase of spleen index and thymus index of infected mice, reduce back skin lesion score of the infected mice, reduce content of IL-6, IL-33 and TNF-alpha in serum of the infected mice, and has a good treatment effect on the specific dermatitis.

To provide a microbial cell material having excellent property.SOLUTION: This is a dead cell body of bacteria from the Blautia genus, characterized by being obtained by heat treatment of Blautia genus bacteria under acidic conditions. Additionally, it is a dead cell body of Blautia genus bacteria, where the IL-12 production induction capability measured using mouse spleen cells is more than ten times greater compared to the IL-12 production induction capability by live Blautia genus bacteria under the same conditions. It also refers to a method for producing the dead cell body of Blautia genus bacteria, characterized by subjecting live Blautia genus bacteria to heat treatment under acidic conditions.SELECTED DRAWING: None

This invention provides a monoclonal antibody against the VP2 protein of a mandarin fish two-segmented RNA virus, its preparation method, and its application. The VP2 protein was obtained by expression and purification using recombinant plasmids. Mice were immunized three times, and mice with the highest serum titer were selected for intraperitoneal booster immunization against VP2 protein. Mouse spleen cells and SP2 / 0 myeloma cells were fused, and hybridoma cell lines secreting VP2 protein antibodies were screened. These cells were then subcloned to obtain four stable hybridoma cell lines secreting anti-VP2 protein antibodies, and their heavy and light chain variable region sequences were determined. Mice were injected intraperitoneally with Freund's incomplete adjuvant, followed by the injection of the hybridoma cell lines. Ascites fluid was collected from the mice, extracted, purified, and used to obtain the monoclonal antibody against the VP2 protein. Using these four monoclonal antibodies, three highly conserved and specific antigenic epitopes of the VP2 protein of the mandarin fish two-segmented RNA virus were screened and identified. This invention provides support for establishing specific serological methods for mandarin fish two-segmented RNA viruses and for further research on the biological function and clinical diagnosis of VP2.

The invention relates to the technical field of collagen peptides, in particular to a collagen peptide capable of enhancing immunity and a preparation method thereof.The preparation method comprises the steps that squid skin is subjected to enzymolysis under the alkaline condition to obtain enzymatic hydrolysate, pitch-based activated carbon is added into the enzymatic hydrolysate for decoloration and deodorization, and then low-temperature freeze drying is conducted. The collagen peptide disclosed by the invention can effectively improve spleen and thymus indexes of mice, remarkably improve the phagocytic rate and phagocytic index of macrophages, play a role in enhancing immunity, and have wide application prospects in the fields of health foods and medicines.

Isolated peptides capable of reducing the amount of dexamethasone-induced spleen and / or thymus weight loss in a mouse are disclosed as well as uses thereof. 20 24 27 84 77 16 D ec 2 02 4 A B S T R A C T 2 0 2 4 2 7 8 4 7 7 1 6 D e c 2 0 2 4

The invention belongs to the technical field of biology, and relates to an in-vitro preliminary screening method of an IL-6 / JAK2 / STAT3 signal channel inhibitor. The method comprises the following steps: co-culturing mouse spleen lymphocytes and a to-be-detected compound, and determining the critical concentration of the to-be-detected compound which does not reduce the cell activity; dividing the mouse spleen lymphocytes into a model group, an experimental group and a control group; only 10-20ng / mL of IL-6 is added into the culture solution of the model group, 10-20ng / mL of IL-6 and a to-be-detected compound with critical concentration are added into the culture solution of the experimental group, and no additional substance is added into the culture solution of the control group; if the STAT3 mRNA expression quantity, the STAT3 and p-STAT3 protein expression quantity and the IgG content of the model group are all higher than those of the control group and the experimental group, the compound to be detected is judged to be the IL-6 / JAK2 / STAT3 signal channel inhibitor. The method can reduce resource investment of animal experiments and clinical tests.

The invention belongs to the technical field of biology, and particularly relates to application of a pulse ultrasound instrument in preparation of a medical instrument for improving the immunocompetence of T lymphocytes. The application creatively discovers for the first time that when the T lymphocytes of the spleen of the mouse are irradiated by adopting a pulse ultrasound instrument, endogenous IL-2 secretion can be promoted, PD-1 expression can be reduced, the immunocompetence of the T lymphocytes can be enhanced, the tumor killing effect of CD4 + and CD8 + effector T lymphocytes can be improved, and a new means is provided for tumor treatment.

The present invention discloses a monoclonal antibody specifically binding to Staphylococcus aureus manganese ion transporter C and its application. The antibody MntC-15 provided by the present invention can specifically bind to the MntC protein, can effectively inhibit the growth of Staphylococcus aureus, can resist sepsis caused by MRSA infection, and inhibit the release of mouse spleen lymphocyte cytokines induced by MntC, and has broad application prospects in clinical practice.

This invention provides a neutralizing monoclonal antibody 3E3 against the hemagglutinin protein of H7 subtype influenza virus and its applications. This antibody specifically recognizes the hemagglutinin protein of H7 subtype influenza virus and exerts an antiviral effect through neutralization. The hybridoma cells that produce this monoclonal antibody are obtained by fusing BALB / C mouse spleen lymphocytes immunized with purified hemagglutinin protein and mouse myeloma cells SP2 / 0 through a process of cloning, selection, and passage, ultimately resulting in cells that stably secrete antibody 3E3. The amino acid sequence of the heavy chain variable region of the antibody is shown in SEQ ID No. 2, and the amino acid sequence of the light chain variable region is shown in SEQ ID No. 4. 3E3 can effectively bind to, neutralize, and treat H7 subtype influenza virus infection.