41 results about "Virus infected cell" patented technology

The invention discloses an Nfkb1 gene overexpression cell line and a construction method thereof, and relates to the field of gene engineering. The constructed recombinant lentiviral expression vector comprises a first LTR, a CMV promoter, an Nfkb1 gene, a Flag polypeptide tag, an EF-1a promoter, a green fluorescent protein gene, a shearing element, a screening gene and a nucleic acid fragment of a second LTR which are sequentially arranged from the 5'end to the 3 'end; co-transfecting a host cell by using the recombinant lentivirus expression vector and lentivirus packaging helper plasmid, and carrying out virus packaging to obtain an Nfkb1-overexpressed recombinant lentivirus; the HT22 cell line overexpressing the Nfkb1 is obtained by infecting the HT22 cell with the recombinant lentivirus overexpressing the Nfkb1, and screening the HT22 cell line overexpressing the Nfkb1 by using a resistance gene. Recombinant lentivirus is used as a vector, HT22 cells are infected with the recombinant lentivirus vector, a cell line capable of stably overexpressing the Nfkb1 gene is obtained through screening, and the effect of the Nfkb1 in neuroinflammation is conveniently researched.

The invention discloses a nano antibody for resisting a novel coronavirus mutant strain RBD structural domain antigen. The amino acid full-length sequence of the nano antibody is shown as any one of SEQ ID NO.5-8. The nano antibody disclosed by the invention has good capability of blocking the combination of a novel coronavirus mutant strain RBD and a receptor thereof, and can effectively prevent cells from being infected by viruses through experimental verification.

The invention discloses an application of Rottlerin in preparation of a medicine for preventing and treating nervous necrosis virus. According to the application disclosed by the invention, the fact that the rotinidin has high binding energy to RGNNV capsid protein is found, the rotinidin without toxic concentration is selected to carry out an RGNNV virus infected cell experiment, and the result shows that the rotinidin inhibits the replication of the RGNNV virus in a dose-dependent manner. The safe dosage of Rottlerin is obtained according to the survival rate of zebrafish treated with different doses, the influence of Rottlerin on the survival rate of zebrafish infected with RGNNV is researched, and finally it is proved that Rottlerin has a remarkable treatment effect on zebrafish infected with RGNNV. Rottlerin provides a new natural small molecule drug choice for prevention and treatment of VNN diseases, and has important research and development value and development significance for prevention and treatment of nervous necrosis viruses.

The invention belongs to the field of biological medicine, and relates to a CAR-T cell targeting NKG2DL and secreting IL-15 and a preparation method thereof, the method comprises the step of infecting a T cell with a recombinant lentivirus, and the recombinant lentivirus contains a nucleic acid molecule of a chimeric antigen receptor encoding NKG2D and IL-15 or is prepared through an expression vector containing the nucleic acid molecule. The CAR-T cell expresses a chimeric antigen receptor targeting NKG2DL and can secrete a cell factor IL-15, the proliferation and effector functions of the CAR-T cell are promoted, and the killing activity on tumors is enhanced.

Provided are a Dabie bandavirus mRNA vaccine and a preparation method therefor. Specifically an optimized mRNA molecule is provided, and is cloned into a pGEM-3Zf(+)mRNA vaccine vector; the plasmid is linearized by means of enzyme digestion, and subjected to capping and poly(A) tail addition by means of an in-vitro transcriptase method to prepare an mRNA; the obtained mRNA is transfected into eukaryotic cells, and it is verified by means of an immunoblotting experiment that the mRNA can be expressed in vitro; and the mRNA is encapsulated by a lipid nanoparticle delivery system to obtain the mRNA vaccine, which, after immunizing mice, can induce the generation of a relatively high antibody level in serum. A viral neutralization test further proves that the immune serum can bind to viruses to prevent the viruses from infecting cells.

The invention belongs to the field of biological medicine, and relates to a CAR-T cell targeting NKG2DL and secreting IL-10 and a preparation method thereof, the method comprises the step of infecting a T cell with a recombinant lentivirus, and the recombinant lentivirus contains a nucleic acid molecule of a chimeric antigen receptor encoding NKG2D and IL-10 or is prepared through an expression vector containing the nucleic acid molecule. The CAR-T cell expresses a chimeric antigen receptor targeting NKG2DL and can secrete a cell factor IL-10, the proliferation and effector functions of the CAR-T cell are promoted, and the killing activity on tumors is enhanced.

To provide methods and compositions for reducing virus titer and eliminating virus-infected cells from an individual.SOLUTION: Provided is the use of an anti-CD24 antibody in the production of medicament for use in a method for reducing hepatitis virus in an individual, where the method comprises: contacting the individual to a therapeutically effective dose of the anti-CD24 antibody for a period of time sufficient to reduce the amount of hepatitis virus, where the hepatitis virus is one or more of hepatitis B virus (HBV) and hepatitis delta virus (HDV).SELECTED DRAWING: None

Disclosed herein is a small molecule targeted drug conjugate for anti-influenza chemotherapy and immunotherapy. The disclosed drug conjugate may form an adaptor to recruit additional CAR T cells or other immune cells for precise elimination of influenza virus-infected cells in a subject. Concurrently administered antibodies or pre-existing immunity in influenza-virus infected subject works well with the targeted conjugate to eliminate virus infected cells, saving valuable time for rescuing late stage patients.

The application provides a benzene [d] isoxazole compound and an application thereof. The compound has the structure shown in the following formula I. The benzene [d] isoxazole compound provided by the application can induce degradation of BET protein and / or GSPT1 protein, and has an anti-proliferation effect on cancer cells. Therefore, the compound and the composition provided by the application can be used for preparing a drug for treating or preventing tumor formation, inflammation, viral infection, cell proliferative disorder, autoimmune disease, sepsis and the like.

The invention belongs to the technical field of virus infection cell lines, and particularly relates to a rats kidney cell line and application thereof. Wherein the preservation number of the kidney cell line of the rats with the yellow chest is CCTCC (China Center For Type Culture Collection) NO: C2025271. The rats kidney cell line (RtK-15W) provided by the invention has multiple advantages of high sensitivity, high replication efficiency, quantitative detection, passage stability and the like on arenavirus; a reliable cell tool and an efficient, reliable and generalizable experimental platform are provided for separation and identification, in-vitro amplification, titer determination, pathogenesis research, drug screening, antibody neutralization experiment, vaccine effect evaluation and the like of arenaviruses (including lymphocytic choriomeningitis virus (LCMV) and Wenzhou virus (WENV)).

The application discloses a kind of targeted exosome chimeric antigen receptor molecules for treating HIV infection and its application, belong to the field of biological medicine technology.The application is packaged into targeted exosome by coding anti-HIV CAR mRNA directionally.The nano antibody sequence of targeted CD3, CD4, CD8+T receptor on T cell is fused to the N-terminal of LAMP-2B to construct specific targeting plasmid, so that the targeted peptide is expressed on the surface of exosome membrane, the expression plasmid containing RNA binding protein L7A and exosome tag protein CD63 is constructed, and the exosome loaded with CAR mRNA and targeted to T cell is obtained by three-plasmid co-transfection of HEK293F cells containing C / D box CAR plasmid.The exosome can construct human CAR-T cell when incubated with PBMC in vitro or back into humanized mouse in vivo, and has significant killing virus infected cell activity.

The present disclosure relates to the field of methods for culturing virally-infected cells. More particularly, this disclosure relates to methods of culturing cells with an influenza A virus. The disclosure relates further to impact of certain culture factors such as trypsin concentration, pH levels, incubation time, and combinations thereof on the production of viral proteins, particularly haemagglutinin.

[Problem] To provide an antiviral oligonucleotide that acts on cells infected by viruses having multiple RNA genomes, exhibiting an antiviral effect, and does not tend to the emergence of resistant viruses, and a pharmacological composition thereof. [Solution] An oligonucleotide that takes on a G-quadruplex structure that inhibits the binding of the genomic RNA and nucleocapsid (N) protein of a virus having an RNA genome, the antiviral oligonucleotide suppressing proliferation of the virus by contact with virus-infected cells, and a pharmacological composition thereof are used.

The invention provides a PK-15 cell line capable of stably expressing porcine Mx2 protein as well as preparation and application of the PK-15 cell line. The PK-15 cell line capable of stably expressing porcine Mx2 gene is constructed through a lentiviral vector system. A specific primer is designed according to the nucleotide sequence of the porcine Mx2 gene in a GeneBank database, recombinant lentivirus plasmids are constructed after target genes are amplified, HEK293T cell packaging lentivirus is transfected by utilizing a three-plasmid system, PK-15 cells are infected by the obtained recombinant lentivirus, and the PK-15 cell line for stably expressing the porcine Mx2 gene is obtained through puromycin screening. Experimental results show that the PK-15 cell line PK-15-poMx2 capable of stably expressing the porcine Mx2 protein is successfully constructed by the method disclosed by the invention. Compared with normal PK-15 cells, the expression quantity of Mx2 in the porcine Mx2 stable expression cell line is remarkably increased, and the porcine Mx2 stable expression cell line has an inhibiting effect on replication of PRV (Porcine Reproductive Virus). Therefore, the successful construction of the cell line for stably expressing the porcine Mx2 protein provides an important tool for deeply exploring the functional characteristics and the antiviral action mechanism of the porcine Mx2 protein.

Methods for providing a vaccine composition capable of effectively inducing a systemic immune response and / or a topical immune response upon administration wherein the composition comprises a human leukocyte antigen class I (HLAI) restrictive epitope selected from the viral pathogen non-virion constituent protein (non-VIP) and thus elicits a CD8T cell response specific against viral infected cells.

A method for providing a vaccine composition capable of effectively inducing a systemic immune response and / or a local immune response upon administration, wherein the composition comprises a human leukocyte antigen class I (HLA I)-restricted epitope selected from viral pathogen non-viral particle constituent proteins (non-VIPs), and thus elicits a CD8 T cell response specific for virus-infected cells.

The application belongs to the technical field of biological medicine, and provides a recombinant type III human collagen peptide for resisting HPV infection and a preparation method and application thereof, an amino acid sequence of which is a coding sequence of positions 991-1032 of type III human collagen UniProtKB: P02461 repeated three times, named as ATHPV3, and an amino acid sequence of which is shown as SEQ ID NO. 1. After expression and purification, the recombinant type III human collagen peptide with a molecular weight of about 13 kD is obtained, can be specifically combined with HPV 16 / 18 L1 protein, has multiple hydrogen bond actions, and can inhibit pseudo-virus infection of 293T cells at high and medium concentrations. The application successfully obtains a recombinant type III human collagen peptide with good purity, which can be combined with HPV 16 / 18 L1 and can effectively inhibit HPV 16 / 18 pseudo-virus infection of 293T cells, and provides an effective candidate drug for preventing and treating HPV infection.

The application provides a kind of 2,6-piperidinedione derivatives and preparation method and application thereof.The compound has the structure shown in the following formula I, formula II or formula III:2,6-piperidinedione derivatives described in the application can induce the degradation of GSPT1 protein, and have cytotoxic effect on cancer cells.Therefore, the compound and composition provided by the application can be used for preparing drugs for treating or preventing tumor formation, inflammation, viral infection, cell proliferative disorder, autoimmune disease, sepsis and other related diseases.

The invention provides a preparation method of a lymphoma autologous tumor vaccine with high expression of TRIM-21, which comprises the following steps: amplifying a TRIM-21 gene from EL-4 cells through a PCR (Polymerase Chain Reaction) method, cloning the TRIM-21 gene into a lentiviral vector to construct a recombinant lentivirus, infecting the EL-4 cells with the recombinant lentivirus to obtain a lymphoma cell line with high expression of TRIM-21, and carrying out irradiation inactivation to obtain the lymphoma autologous tumor vaccine with high expression of TRIM-21. The lymphoma autologous tumor vaccine is obtained. The lymphoma cell line with high expression of TRIM-21 is prepared for the first time, and the lymphoma autologous tumor vaccine is prepared from the lymphoma cell line. A mouse tumor model verifies that the vaccine efficiently activates CD8 + T cell immune response and has an obvious tumor immunoprophylaxis effect, so that the vaccine can be applied to the field of preparation of tumor immunoprophylaxis drugs.

A method of culturing cells for vaccine production comprises culturing cells in the presence of microcarriers with cell culture media and infecting the cells with a virus. The microcarrier comprises a bead and a coating. The coating comprises a thermo-responsive polymer having a lower critical solution temperature (LCST) of between about 20 °C and about 34 °C. The cells adhere to the coating of the microcarrier at a temperature above the LCST. The microcarrier comprises a polymeric bead, and a hydrophobic polymer. The hydrophobic polymer is a block copolymer which is connected to the bead by a covalent bond or by physical adsorption. The block copolymer comprises at least one hydrophobic block and at least one thermo-responsive block.

The application provides a polypeptide molecule for specifically recognizing HIV gp120 protein and an application thereof, and belongs to the technical field of biological medicine. The application provides a lead polypeptide derivative for specifically targeting and combining HIV gp120, wherein the lead polypeptide comprises an amino acid sequence shown in SEQ ID No. 1. The application also provides a lead polypeptide derivative prepared based on the lead polypeptide, and a polypeptide nanoparticle constructed by the lead polypeptide derivative, and both of them have gp120 protein combining capacity. The derivative and the nanoparticle have good virus neutralization inhibition effect. The series of polypeptide molecules can specifically combine with free HIV virus, HIV infected cells and free gp120, so as to block HIV infected cells, delay the AIDS disease process, realize AIDS treatment drug targeted delivery, detection and diagnosis.

The invention discloses a reagent and a kit for detecting EB (Epstein-Barr) virus infection and a method for detecting EB virus infected cell lineage, and belongs to the field of medical diagnosis. The technical problem to be solved by the invention is how to accurately locate different cell lineages of EBER + cells. The reagent for detecting EB virus infection disclosed by the invention contains an EBV probe and an antibody for marking a cell lineage, the sequence of the EBV probe is shown as SEQ ID No.1 and SEQ ID No.2, and the antibody for marking the cell lineage is a CD3 antibody, a CD20 antibody and / or a CD56 antibody. The reagent and the kit for detecting EB virus infection can be used for simultaneous marking of EBER genes and antigen proteins, accurate positioning of different cell lineages of EBER + cells is achieved, and cell lineage positioning of EB virus infection is further achieved.

This invention belongs to the field of virus-infected cell line technology, and particularly relates to a striped field mouse lung cell line and its applications. The striped field mouse lung cell line has the accession number CCTCC NO: C2025268. The AaL-09W cell line (including primary and passaged cells) provided by this invention has multiple advantages, including high sensitivity to Hantavirus, high replication efficiency, high isolation success rate, closer resemblance to the host's natural environment, passage stability, and easy scaling-up. It provides a reliable cellular tool and an efficient, reliable, and scalable experimental platform for the isolation, identification, in vitro culture, titer determination, infection mechanism and pathogenesis research, new drug screening, and vaccine development of Hantavirus, meeting the needs of efficient culture and multi-scenario applications of HTNV and related Hantaviruses.

A human-derived SIRT6 gene is amplified and cloned to a plasmid vector to construct a pSIRT6 plasmid, and under the action of Sirt6, the proportion of memory CD8 + T cells causing mitochondrial dysfunction after SARS-CoV-2S protein stimulation of a new coronavirus is reduced, so that the number of normal memory CD8 + T cells is increased. A further research shows that the protein phosphorylation levels of PI3K, Akt and mTOR in the CD8 + T cells stimulated by the SARS-CoV-2S protein are inhibited, and the whole phosphorylated protein level in the cells is reduced. For a constructed mouse influenza A virus infection model, the virus load is reduced under the action of Sirt6, the inflammatory response of a mouse lung pathological section is obviously reduced, the generation of memory CD8 + T cells in a mouse PBMC (peripheral blood mononuclear cell) is increased, and the mitochondrial function of the memory CD8 + T cells gradually returns to normal. The invention records that the human STRT6 gene has a remarkable treatment effect on the cellular immune process caused by the new coronavirus and the influenza virus.

The application discloses a compound with a structural formula I, wherein the compound has good 3CL protease inhibitory activity, has significant proliferation inhibitory activity on virus-infected cells, and has potential application value in treating diseases related to virus infection. The compound has good solubility and permeability, good in-vivo metabolic stability, high in-vivo exposure, and high bioavailability, and is a potential drug compound.

The invention discloses a method for predicting a CAR-T killing effect in vitro based on NO probe cell imaging, and relates to the field of immunotherapy curative effect evaluation. The method comprises the following steps: detecting the expression quantity of CD58 in cells through virus infection, a protein imprinting experiment and a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) experiment, then incubating tumor cells in vitro through an NO fluorescent probe, imaging to detect a fluorescent signal, finally co-culturing CAR-T and the tumor cells in vitro, and evaluating the killing effect through flow cytometry. The expression level of CD58 is reflected through the intensity of a fluorescence signal generated when the NO probe acts on tumor cells, and the killing effect of CAR-T is reflected through the expression level of CD58. According to the invention, the NO probe technology is utilized for simple, sensitive, rapid and visual detection, and the killing activity of the CAR-T is evaluated in vitro, so that a basis is provided for follow-up in-vivo killing treatment of the CAR-T.

The invention provides a high-throughput screening method of an anti-Nipah virus drug. The method comprises the following steps: firstly, completing drug gradient dilution and planking pretreatment in a BSL-2 laboratory; then carrying out virus infection, cell culture and 72-hour incubation in the BSL-4 core region; the method comprises the following steps: carrying out rapid and low-cost visual preliminary screening on a whole-plate sample through crystal violet dyeing, and selectively carrying out TRIzol LS inactivation treatment on a small amount of potential effective sample supernatant; after the inactivated sample is transferred to a BSL-2 laboratory, the virus copy number is accurately quantified by adopting a qRT-PCR technology, and finally the drug inhibition ratio and the EC50 value are calculated. According to the method, through a staged screening strategy, on the premise of ensuring biological safety, the screening flux is greatly improved, the detection cost is reduced, and a reliable solution is provided for drug screening of high-pathogenicity pathogenic microorganisms.

The embodiment of the invention provides a macrophage overexpressing IL-10, and further provides a preparation method of the macrophage overexpressing IL-10, and application of the macrophage overexpressing IL-10 in preparation of a product for preventing or treating atherosclerosis. According to the invention, RAW264.7 cells are infected by packaging recombinant lentivirus overexpressing mouse source IL-10, the macrophage overexpressing IL-10 is finally prepared, and the macrophage overexpressing IL-10 can reduce the plaque area and the plaque necrosis core and increase the plaque stability when intervening AS. The macrophage overexpressing IL-10 is expected to be applied to clinical prevention and / or treatment of atherosclerosis, reduction of ischemic cardiovascular disease burden and reduction of occurrence of cardiovascular events.

The present application belongs to the field of prevention and control of aquatic pathogenic microorganisms, and particularly relates to a fish rhabdovirus binding protein and a preparation method and application thereof. The protein can efficiently bind to the site of fish rhabdovirus binding to a host cell receptor, effectively block the invasion channel of fish rhabdovirus invading a host cell, thereby blocking virus infection of a cell and avoiding virus replication and proliferation in a host body. The fish rhabdovirus binding protein provided by the present application has a high-efficiency anti-rhabdovirus effect, and a premixed feed additive prepared by taking the protein as an active material can significantly improve the anti-rhabdovirus capacity of fry, is safe and non-polluting, can be applied on a large scale, and can further improve the economic benefits of fish culture.

The invention discloses an application of cis-BBP in inactivation of viruses with envelope structures. Any one of the following applications of the cis-BBP comprises an application of the cis-BBP in photodynamic inactivation of the virus with the envelope structure, an application of the cis-BBP in preparation of a product for preventing and treating the virus with the envelope structure, and an application of the cis-BBP in preparation of a product for inhibiting cells infected by the virus with the envelope structure. According to the application of the cis-BBP in photodynamic inactivation of the virus with the envelope structure, the antiviral efficiency is high, and a broad-spectrum antiviral strategy for the virus with the envelope structure is developed. The broad-spectrum antiviral ability of the cis-BBP is explored by utilizing the characteristics of the cis-BBP, and the action effect of the cis-BBP on various viruses is clarified. The cis-BBP shows antiviral activity and excellent biocompatibility in vivo. When applied to transportation and packaging, it shows significant potential in controlling virus propagation.