197 results about "Splenocyte" patented technology

The invention provides a rabbit-derived immortal B-lymphocyte capable of fusion with a rabbit splenocyte to produce a hybrid cell that produces an antibody. The immortal B-lymphocyte does not detectably express endogenous immunoglobulin heavy chain and may contain, in certain embodiments, an altered immunoglobulin heavy chain-encoding gene. A hybridoma resulting from fusion between the subject immortal B-lymphocyte and a rabbit antibody-producing cell is provided, as is a method of using that hybridoma to produce an antibody. The subject invention finds use in a variety of different diagnostic, therapeutic and research applications.

The present invention is a recombinant vector encoding and expressing at least three or more costimulatory molecules. The recombinant vector may additionally contain a gene encoding one or more target antigens or immunological epitope thereof. The synergistic effect of these costimulatory molecules on the enhanced activation of T cells is demonstrated. The degree of T-cell activation using recombinant vectors containing genes encoding three costimulatory molecules was far greater than the sum of recombinant vector constructs containing one costimulatory molecule and greater than the use of two costimulatory molecules. Results employing the triple costimulatory vectors were most dramatic under conditions of either low levels of first signal or low stimulator to T-cell ratios. This phenomenon was observed with both isolated CD4+ and CD8+ T cells. The recombinant vectors of the present invention are useful as immunogenes and vaccines against cancer and pathogenic micro-organisms, and in providing host cells, including dendritic cells and splenocytes with enhanced antigen-presenting functions.

The present invention is a recombinant vector encoding and expressing at least three or more costimulatory molecules. The recombinant vector may additionally contain a gene encoding one or more target antigens or immunological epitope thereof. The synergistic effect of them costimulatory molecules on the enhanced activation of T cells is demonstrated. The degree of T-cell activation using recombinant vectors containing genes encoding three costimulatory molecules was far greater than the sum of recombinant vector constructs containing one costimulatory molecule and greater that the use of two costimulatory molecules. Results employing the triple costimulatory vectors were most dramatic under conditions of either low levels of first signal or low stimulator to T-cell ratios. This phenomenon was observed with both isolated CD4+and CD8+T cells. The recombinant vectors of the present invention are useful as immunogenes and vaccines against cancer and pathogenic micro-organisms, and in providing host cells, including dendritic cells and splenocytes with enhanced and antigen-presenting functions.

An exemplary embodiment providing one or more improvements includes feeding animals with probiotic microbes encapsulated in a mixture of xanthan gum and chitosan, or in gelatin, specifically Pediococcus acidilactici and Saccharomyces boulardii. Such encapsulation protects the viability of the probiotic microbes against unfavorable temperatures. An exemplary embodiment providing one or more improvements includes methods of using viable probiotics in therapy of birds and mammals infected with infectious diseases. Probiotics acted as adjuvants in stimulating antibody reaction and stimulated a cellular immunity response. In particular, probiotics were shown to reduce the number of viable oocytes from fecal samples, stimulate antibody production, and stimulate of proliferation of splenocytes in chickens infected with Elimeria. In addition, probiotics were shown to relieve symptoms of parvovirus infection in dogs.

An exemplary embodiment providing one or more improvements includes feeding animals with probiotic microbes encapsulated in a mixture of xanthan gum and chitosan, or in gelatin, specifically Pediococcus acidilactici and Saccharomyces boulardii. Such encapsulation protects the viability of the probiotic microbes against unfavorable temperatures. An exemplary embodiment providing one or more improvements includes methods of using viable probiotics in therapy of birds and mammals infected with infectious diseases. Probiotics acted as adjuvants in stimulating antibody reaction and stimulated a cellular immunity response. In particular, probiotics were shown to reduce the number of viable oocytes from fecal samples, stimulate antibody production, and stimulate of proliferation of splenocytes in chickens infected with Elimeria. In addition, probiotics were shown to relieve symptoms of parvovirus infection in dogs.

The invention discloses a preparation method of an anti-bisphenol A monoclonal antibody. The preparation method comprises the following steps of: combining bisphenol A and a macromolecule carrier protein to prepare an artificial immunity antigen and a coating antigen; preparing splenocyte suspension from a bisphenol A artificial immunity antigen immunity mouse; fusing the mouse splenocyte and mouse myeloma cells SP2 / 0 in the presence of polyethylene glycol; detecting by hybridoma technology and subcloning for many times to obtain positive hybridoma cells which can stably secrete bisphenol A antibody, wherein the secreted antibody belongs to the IgG1 subclass; and establishing a bisphenol A indirect competition ELISA (Enzyme-Linked Immunosorbent Assay) test by utilizing the monoclonal antibody secreted by cell lines with high antibodytiter, wherein the minimum limit of detection can reach 0.324ng / mL. The antibody has no cross reaction with compounds with benzene ring structures, such as benzene, phenol, tert-butylphenol, p-hydroxylphengl, o-hydroxybenzoic acid and the like, has strong specificity and can be used for immunity detection of bisphenol A.

A novel representational system that is a method for improving feeding probiotics is provided. The probiotics, especially the Pediococcus acidilactici and the Saccharomyces boulardii are mixed in different colloid net including xanthan, chitosan or gelatin. By using said mixed colloid the activity of the probiotics can be protected, and multiple execrable surroundings and disadvantageous temperature for growth can be prevented. The novel representational method also provides many kinds of method for treating infectious disease for birds and mammals by using activated probiotics. Furthermore, the probiotics is usually acted as adjuvants for improving the generation of antibody and the cellularity immunity effect. In particular, probiotics were shown to reduce the number of viable oocytes from fecal samples, stimulate antibody production, and stimulate of proliferation of splenocytes in chickens infected with Elimeria. In addition, probiotics were shown to relieve symptoms of parvovirus infection in dogs.

The present invention is a recombinant vector encoding and expressing at least three or more costimulatory molecules. The recombinant vector may additionally contain a gene encoding one or more target antigens or immunological epitope thereof. The synergistic effect of these costimulatory molecules on the enhanced activation of T cells is demonstrated. The degree of Tell activation using recombinant vectors containing genes encoding three costimulatory molecules was far greater than the sum of recombinant vector constructs containing one costimulatory molecule and greater than the use of two costimulatory molecules. Results employing the triple costimulatory vectors were most dramatic under conditions of either low levels of first signal or low stimulator to T-cell ratios. This phenomenon was observed with both isolated CD4+ and CD8+ T cells. The recombinant vectors of the present invention are useful as immunogenes and vaccines against cancer and pathogenic micro-organisms, and in providing host cells, including dendritic cells and splenocytes with enhanced antigen-presenting functions.

The invention provides a new cell sorting magnetic bead, which is a 50-70 nm gold plated Fe3O4 magnetic nanoparticle (Fe3O4 and Au nanoparticle) ostensibly coupled with proteinA by covalent bonds, interacted with the specificity of a Fc section of IgG through the proteinA, and directionally coupled with IgG type mouse CD3 antibodies. The invention relates to a method for synthesizing and modifying the magnetic bead, and a method for carrying out cell sorting by using the magnetic bead. The magnetic bead of the invention is used for removing the CD3 T cells in the mouse splenocytes so as to reduce the CD3 T cell content of the splenocytes from 36.5 percent to 0.7 percent, and has extremely high sorting efficiency. The magnetic bead also has the advantages of high protein-coupled quantity and superparamagnetism, and the preparation method has the advantages of simpleness, environmental protection, low cost and good application prospect.

The invention discloses a making method of heavy-metal lead monoclonal antibody in the biological technical domain, which is characterized by the following: coupling lead ion and carrier protein through p-aminobenzene-diethylenetriamine pentoacetic acid as bifunctional metal chelant; obtaining full antigen; crossing splenocyte and Sp2 / 0 marrow tumour cell to make crossed tumour cell; sieving; obtaining stable strong secretory specificity anti-lead monoclonal monomer to inhibit Pb-DTPA.

The invention provides methods for treating pathological conditions associated with an undesirable inflammatory component. The invention is generally directed to reducing inflammation by administering cells that have one or more of the following effects in an injured subject: interact with splenocytes, preserve splenic mass, increase proliferation of CD4+ and CD8+ T-cells, increase IL-4 and IL-10, decrease IL-6 and IL-1β, and increase M2:M1 macrophage ratio at the site of injury. The invention is also directed to drug discovery methods to screen for agents that modulate the ability of the cells to have these effects. The invention is also directed to cell banks that can be used to provide cells for administration to a subject, the banks comprising cells having desired potency for achieving these effects.

The invention discloses a carbendazim monoclonal antibody, a preparation method and an application thereof. The carbendazim monoclonal antibody is prepared by a mouse hybridoma cell strain MC-3 with a preservation number of CGMCC N0.4575 and the preparation method is as follows: (1) introducing ninth amino acid residue of carbendazim into one amino group to obtain the carbendazim modified by the amino and coupling the carbendazim modified by the amino with a carrier protein to obtain a complete antigen A; (2) immunizing a mouse by the complete antigen A obtained in step (1), taking splenocyteof the immune mice to fuse with a mouse myeloma cell to screen out a positive hybridoma cell strain; extensively culturing the positive hybridoma cell strain and injecting the positive cell strain into a homologous mouse abdominal cavity to induce to generate ascitic fluid and to obtain more carbendazim monoclonal antibodies. The Carbendazim monoclonal antibody can be used for detecting the carbendazim.

The invention discloses a monoclonal antibody preparing method of high-flux, which comprises the following steps: (a) proceeding immune inoculation for non-human mammal animal through multiple immunogen; (b) compounding splenocyte of immune inoculation animal and bone marrow oncocyte to fuse into cross oncocyte; (c) collecting the cross oncocyte; culturing; preparing cross oncocyte suspension; (d) screening cross oncocyte through biological chip to produce cross oncocyte system of immunogen monoclonal antibody.

Leptin was previously demonstrated to exert potent immune modulatory properties in several immune mediated disorders. The aim of the study was to determine leptin's anti-tumor effect in a murine model of human hepatocellular carcinoma (HCC). In vivo, Athymic T cell deficient (nude) mice transplanted with 1×106 human Hep3B cells, followed by administration of two daily intraperitoneal doses of 0.5 mg / gram leptin for 6 weeks. Leptin administration induced a significant reduction in tumor size and improved survival in nude mice. Histologically, tumors of leptin-administered mice featured increased inflammatory exudate in interphase areas. Leptin-induced tumor suppression was associated with a significant increase in peripheral natural killer (NK) cell number. Splenocytes from leptin-treated mice featured decreased expression of CIS mRNA. To determine which lymphocyte subset is a prerequisite for the anti tumor effect of leptin, T&B cell deficient (Scid) mice and T,B& NK deficient (Scid-Beige) mice were subcutaneously implanted with Hep3B tumor cells, with and without the daily intraperitoneal administration of 0.5 mg / gram leptin for 6 weeks. SCID mice featured leptin-associated tumor suppression similar to those of nude mice. In contrast, NK-deficient SCID-Beige mice developed larger tumors. To further establish natural killer cell's central role in mediation of leptin's anti-tumor effect, NK cells were incubated in vitro with increasing doses of leptin, demonstrating a dose-dependent increase in cytotoxic activity. Incubation of leptin with hepatoma cell line was found to induce a dose-dependent reduction in hepatoma cell proliferation, suggesting an additive direct anti-tumor effect. Further synergism in inhibition of hepatoma cell proliferation in vitro was achieved following addition of natural killer cells. HCC cells expressed leptin receptor mRNA, while addition of leptin induced increased mRMA expression of STAT2 and SOCS1 on tumor cell lines. Leptin administration induces a significant suppression of human HCC. This effect is mediated by induction of natural killer cell proliferation and activation, and by direct inhibition of tumor growth. Decreased natural killer cell expression of inhibitory CIS protein and over expression of the anti-proliferative STAT2 and SOCS1 proteins in HCC lines may underline both anti cancerous effects of leptin.

The invention provides materials and methods for modulating adenoviral tropism for hepatocytes and other cell types such as splenocytes. It relates to the findings that hypervariable regions (HVRs) of the viral hexon protein interact with the Gla domain of the blood clotting factor FX as part of the infective process in vivo. The invention provides means to disrupt the interaction between hexon and FX, thus reducing infection of hepatocytes and splenocytes, as well as use of targeting agents comprising the Gla domain or a fragment thereof to direct adenoviral vectors to desired target cell or tissue types.

Provided herein are a hybridoma cell line producing monoclonal antibody against foot-and-mouth disease virus (FMDV), the monoclonal antibody therefrom, reagent and kit for ELISA, and immunoassay method. The hybridoma cell line is produced by cell fusion of a parental cell and a myeloma cell line and has the same characteristics as the cell line whose strain designation is CmA40 and deposition number is ATCC (To be Provided). The parental cell is a splenocyte isolated from the spleen of a mouse immunized by an antigen derived from a 3ABC non-structural protein (NSP) of FMDV. The antigen used here is expressed by a prokaryotic cell. The monoclonal antibody produced by the hybridoma cell line can specifically recognize a 3ABC polypeptide and does not cross-react with an antiserum of swine vesicular disease virus.

The present invention relates to a modified CpG oligodeoxynucleotide (ODN) which is prepared by coupling a consecutive sequence of deoxyribothymine (dT) to the 3′-terminus of CpG ODN having immunoregularory function, thereby improving immunoactivity of splenocytes, macrophages and peripheral mononuclear cells, and therefore, can be effectively used as a vaccine adjuvant for preventing and treating hepatitis B or an anti-cancer agent. Since the phosphorothioate CpG ODN having the consecutive sequence of dT at its 3′-terminus shows high activity inducing Th-1 immune response and does not elicit in vivo toxicity with guaranteeing its safety, it can be effectively used as a vaccine adjuvant.

The invention discloses a universal phthalic acid esters monoclonal antibody hybridoma cell strain and application thereof, and belongs to the technical field of food safety immunological detection. Hapten is prepared and is coupled with albumen based on the glutaraldehyde method to obtain phthalic acid dibutyl ester complete antigen, and the phthalic acid dibutyl ester complete antigen and freund's adjuvant are evenly mixed to be injected to immune BALB / c mice in a subcutaneous injection mode; envelope antigen is formed in a synthesis mode based on the diazotization method and is used for screening mouse serum and cell supernatant. The splenocyte of the immune mice is fused with myeloma cells of the mice based on a PEG method, and indirect ELISA, indirect competition ELISA screening and three times of subcloning are carried out to obtain selective group hybridoma cell strain monoclonal cell strain C. The monoclonal cell strain C has certain recognition capability on DEHP and DINP, and the requirement for phthalic acid ester plasticizer immunodetection products in current market can be met.

The invention discloses a recombination Staphylococcus aureus enteritis toxin E with SEQ ID NO.1 amino acid sequence, which is composed of pGEX-4T-1 and SEQ ID NO.2 nucleotide sequence. The invention utilizes hyperantigen activity to accelerate the breeding of splenocyte and inhibit tumour from growing, which is fit for preparing high-purity enterotoxin and hyperantigen agent.

The present invention provides a monoclonal antibody having a pathogenic activity that can induce pemphigus lesion, a peptide specifically recognized by the monoclonal antibody and useful as a therapeutic drug for pemphigus autoimmune disease, etc. As anti-mouse Dsg3 antibody-producing cells are present in the splenocytes of pemphigus vulgaris mouse model constructed by using autoantigen knockout mouse, cell fusion was conducted with the splenocytes of said mouse model and mouse myeloma cells using polyethyleneglycol, hybridomas were constructed, and monoclonal antibodies against Dsg3 were constructed. Among them, a monoclonal antibody having a pathological activity that can induce pemphigus lesions was screened, the base sequence and amino acid sequence in its variable region (heavy chain, light chain) was determined, and the specific epitope part was identified.

A new protein, named ACA1, has been isolated and purified from the medical fungi Antrodia camphorata using the technique of anion-exchange chromatography. ACA1, a glycoprotein with a molecular mass of 29 kDa, has a pI value of pH 5.3 and contains 118 amino acids in its peptide moiety. In addition, ACA1 contains methionine, half-cystine and histidine residues, which are not existent in FIP-fve and Ling Zhi-8. ACA1 is not able to agglutinate red blood cells from human and mouse. Moreover, ACA1 possesses immunomodulatory activities, which are demonstrated by their stimulatory activity toward RAW 264.7 macrophages and mouse splenocytes. ACA1 can directly enhance the production of tumor necrosis factor-alpha and nitric oxide by RAW 264.7 macrophages, and induce cell proliferation and interferon-gamma secretion by mouse splenocytes.

The invention provides a rabbit-derived immortal B-lymphocyte capable of fusion with a rabbit splenocyte to produce a hybrid cell that produces an antibody. The immortal B-lymphocyte does not detectably express endogenous immunoglobulin heavy chain and may contain, in certain embodiments, an altered immunoglobulin heavy chain-encoding gene. A hybridoma resulting from fusion between the subject immortal B-lymphocyte and a rabbit antibody-producing cell is provided, as is a method of using that hybridoma to produce an antibody. The subject invention finds use in a variety of different diagnostic, therapeutic and research applications.

The invention discloses the application of cyclocarya paliurus extract in the preparation of a medicament for preventing and treating leukemia. High-content cyclocarya paliurus extract is obtained by performing the purification treatment combined of water extraction, ethanol extraction, membrane dialysis, and gel chromatographic column on cyclocarya paliurus leaves. The cyclocarya paliurus extract contains 90 to 98 percent of polysaccharide; killing activity detection experiments show that the cyclocarya paliurus extract can be used for improving the activity of the splenocytes for killing human leukemia cells K652 in vitro; the killing multiples can reach 2.47 to the maximum. After granules, tablets, buccal tablets, capsules, pills, powder injection or oral liquid prepared by adding certain additives into the cyclocarya paliurus extract disclosed by the invention are orally taken, the effects of preventing and treating the leukemia can be achieved.

An effective prophylactic mucosal gene expression vaccine (GXV), made up of a cocktail of at least 4 different plasmid DNAs encoding corresponding RSV antigens, coacervated with chitosan to formulate nanospheres. In a murine model of RSV infection, intranasal administration with GXV results in significant induction of RSV-specific antibodies, nasal IgA antibodies, cytotoxic T lymphocytes, and IFN-γ production in the lung and splenocytes. A single dose of GXV induces a drastic reduction of viral titers.

The invention provides application of 1 beta-hydroxy alantolactone in preparation of medicine for preventing and curing rheumatoid arthritis. The structural formula of the 1 beta-hydroxy alantolactone is as follows: the 1 beta-hydroxy alantolactone is used for carrying out animal pharmaceutical effect tests, and results show that the 1 beta-hydroxy alantolactone obviously suppresses attack severity and morbidity of CIA (collagen-induced arthritis). TNF-alpha (tumor necrosis factors alpha), IL-17 (interleukin 17) and IFN-gamma (interferon gamma) in supernate of splenocyte can be reduced, secretion of proinflammatory cytokine TNF-alpha, IFN-gamma and IL-17 is suppressed, and accordingly CIA diseases are prevented. Accordingly, the application can be used for preparing medicine for treating rheumatoid arthritis. Medicine composition in the invention is prepared by the 1 beta-hydroxy alantolactone used as active ingredients and conventional medicine carriers and can be prepared into tablets, dispersible tablets, lozenge, orally disintegrating tablets, sustained-release tablets, capsules, soft capsules, dropping pills, granular formulation, injection, powder injection or aerosol and the like. The application has great clinical application values.

The invention relates to a monoclonal antibody for II type carp herpes virus ORF72 albumen. The monoclonal antibody is secreted by hybridoma of which the preservation number is CCTCC NO: C2016141. The hybridoma is prepared by the steps of immunizing mice with GST-ORF72 recombinant protein as antigen to obtain mice splenocyte, and fusing mice splenocyte with mice myeloma cell to obtain the hybridoma. The invention further provides the application of the monoclonal antibody. The monoclonal antibody for II type carp herpes virus ORF72 albumen has the advantages that the monoclonal antibody for II type carp herpes virus ORF72 albumen is made for the first time and can be used for preparing diagnostic reagent for II type carp herpes virus and anti-II type carp herpes virus medicine.

The invention discloses a triadimefon monoclonal antibody hybridoma cell strain B11S and application thereof, and belongs to the technical field of food safety immunodetection. The triadimefon monoclonal antibody hybridoma cell strain B11S disclosed by the invention is preserved in China General Microbiological Culture Collection Center (CGMCC), is classified and named as a monoclonal cell strain,is preserved on October 14, 2019, and has a preservation number of CGMCC No. 18518. A BALB / c mouse is immunized by a triadimefon complete antigen. High-titer and low-IC50 mouse splenocytes are takenand fused with myeloma cells through a PEG method, and the hybridoma cell strain B11S is obtained through screening of an indirect competitive enzyme-linked immunosorbent assay and three times of subcloning. A monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (IC50 value is 5.25 ng / mL) to triadimefon, provides a raw material for immunodetection of triadimefon residues in foods, and has practical application value.

The invention discloses a thiacloprid and acetamiprid monoclonal antibody hybridoma cell strain GW and application thereof, and belongs to the field of food security immunodetection. Thiacloprid and acetamiprid complete antigen of the strain is uniformly mixed with an equal amount of QuickAntibody-Mouse 5W adjuvant, and is injected to BALB / c mice through leg muscle. The dosage is 100mu g / mouse for the first time of immunization, the dosage is 50[mu]g / mouse for multiple times of intensified immunization, the immunization is implemented at an interval of 21 days, and thiacloprid and acetamiprid complete antigen (25[mu]g / mouse, without adjuvant) is adopted for immunization impact for the last time. Splenocyte of high-potency low IC50 mice is taken and fused with mouse myeloma cells by using a PEG method, indirect competitive inhibition enzyme-linked immunosorbent assay is adopted for screening, and three times of subcloning is implemented, so as to obtain the hybridoma cell strain. A monoclonal antibody secreted from the cell strain has relatively good specificity and detection sensitivity (the IC50 values are 0.1ng / mL and 0.4ng / mL respectively) for thiacloprid and acetamiprid, detection on the residual amounts of thiacloprid and acetamiprid in water, fruits and vegetables and cereals can be achieved, conditions are provided for immunodetection on thiacloprid and acetamiprid residues in food can be provided, and practical use values can be made.