12 results about "Murine cell" patented technology

The invention belongs to the technical field of biological medicine, and relates to pharmaceutical application of an oxidative phosphorylation inhibitor, in particular to application of the oxidative phosphorylation inhibitor in preparation of an immunomodulatory drug, and the immunomodulatory drug has the advantages that the function damage of newborn myeloid-derived suppressor cells caused by antagonism BCG (bacillus calmette guerin) vaccination is inhibited, and the immunomodulatory effect is improved. The compound is used for preventing or treating neonatal immune-related diseases caused by bacillus calmette guerin vaccine inoculation. The invention finds that the BCG can cause damage to the immunosuppression function of the MDSC of the suckling mouse through up-regulation oxidative phosphorylation, and the negative effect caused by inoculation of the BCG can be reversed by using the OXPHOS inhibitor, so that the immunosuppression function of the MDSC of the suckling mouse is successfully recovered in an in-vivo and in-vitro model, and the immune-related adverse reaction caused by inoculation of the BCG is expected to be improved.

The invention discloses an application of a Pik3cb inhibitor in treatment of Sjogren's disease. Experiments prove that the Pik3cb inhibitor TGX-221 can reduce the number, activation and chemotaxis of NOD mouse B cells of a Sjogren's disease model mouse, plasma cell differentiation and antibody secretion, improve the immune microenvironment of the submaxillary gland of the Sjogren's disease model mouse, reduce the infiltration degree of submaxillary gland lymphocytes and promote the salivary secretion function; and after the Pik3cb is over-expressed, the symptom and pathology of the Sjogren's disease are aggravated. Therefore, the Pik3cb inhibitor can be used for treating the Sjogren's disease. The invention provides a new thought for research and development of medicines for treating the Sjogren's disease, and has important application value.

The invention discloses a method for inhibiting an RAW264.7 cell from differentiating into an M1 cell. The method comprises the step of inhibiting the RAW264.7 cell from differentiating into the M1 cell by using miR-143-3p or a simulant thereof. According to the application, miR-143-3p is utilized to transfect and inhibit RAW 264.7 cells of mice, and the miR-143-3p targets and inhibits JAK-STAT and NF-kappa B pathways, so that the differentiation of the RAW 264.7 cells to M1 cells is inhibited, and the polarization effect of the M1 cells is remarkably inhibited.

The invention discloses a mouse NK cell in-vitro culture method and application, and belongs to the technical field of cell culture. The method comprises the following two culture steps: firstly, culturing for 1.5-2.5 days by using an activated culture medium containing an MEMS alpha culture medium, 15-25v / v% of FBS, beta-ME and mIL-2 (optional mIL-15 and mIL-12); and culturing for 16.5-17.5 days by using an amplification culture medium containing an MEMS alpha culture medium, FBS, beta-ME and mIL-2, wherein the cell density is controlled to be less than or equal to 2E < 6 > / mL in the whole culture process. According to the culture method provided by the invention, the amplification efficiency of the mNK cells can be remarkably improved, the initial proliferation speed of a multi-factor activation group is higher, the cells are maintained in a normal form, the transduction efficiency is improved, and sufficient high-activity cells are provided for research of the mNK cells.

The present application relates to the field of bioengineering, and particularly relates to a kind of artificial codon optimization nucleic acid molecule of the red fluorescent protein coding.The present application provides nucleic acid molecule of red fluorescent protein coding, and the red fluorescent protein is mCherry, compared with the existing red fluorescent protein nucleotide sequence, the number of CpG site is increased from 56 to 92, the content of the third position of codon G / C (GC3) is not less than 99%.The nucleotide sequence provided by the present application is the mCherry synonymous codon substitution sequence obtained by combining the increase of the number of CpG sites and the use of optimal codon strategy, which is 1.3-2.4 times higher than the fluorescence intensity of the sequence before modification in mammalian (human / mouse) cells;The present application uses the method of codon optimization to obtain the nucleic acid molecule of the red fluorescent protein that does not exist in nature, which can encode the red fluorescent protein with stronger fluorescence intensity, and promotes the development of live cell imaging.

The invention discloses a construction method and application of an in-vitro depletion type CD8 + T cell mouse model. The construction method comprises the following steps: S1, preparing antibody coating liquid and pre-coating plate holes; s2, separating, extracting and culturing mouse CD8 + T cells; s3, carrying out induction establishment on early-stage depletion type CD8 + T cells; s4, establishment of induction of mid-term depletion type CD8 + T cells; and S5, induction establishment of the late exhaustion type CD8 + T cells. The three-high characteristics of the in-vitro depletion CD8 + T cell model, namely high phenotype fidelity, high cell survival rate and high data reproducibility, are realized for the first time, a standardized platform is provided for immune depletion mechanism research, immune checkpoint drug development and combined treatment strategy optimization, and the method has remarkable industrial application value.

The application provides an application of an Angptl8 neutralizing antibody in preparation of a drug for treating mouse cytokine storm syndrome. The present study proves that angiopoietin-like protein 8 (Angptl8) can promote macrophages to release inflammatory factors such as TNF-alpha, IL-6 and IL-1beta by using molecular biology means. In terms of mechanism, Angptl8 can cause the enhancement of the inflammatory factor storm of the body by promoting the polarization of M1 type macrophages and inhibiting the polarization of M2 type macrophages, and the knockout of Angptl8 can improve this pathological phenomenon. Therefore, we synthesize the Angptl8 neutralizing antibody and inject it into the CSS mouse to combine with the Angptl8 in the blood circulation of the mouse, successfully inhibit the large release of the inflammatory factors of the body, and improve the survival rate of the mouse within 72h to 60%. The application proves that the Angptl8 neutralizing antibody can effectively treat the mouse cytokine storm syndrome, and clarifies the molecular mechanism of the target Angptl8 in regulating the disease process, and provides an effective strategy for treating the CSS patient in the clinic.

The invention belongs to the technical field of medicines, and particularly relates to application of CD47 serving as a target spot to preparation of a medicine for treating sepsis. The invention discovers the application of CD47 in preparation of drugs or pharmaceutical compositions for treating sepsis-related diseases for the first time. The survival rate of a sepsis mouse can be remarkably improved by intervening CD47 expression through the CD47 monoclonal antibody / small molecule inhibitor / siRNA, and the research finds that B cell inhibition of the sepsis mouse is relieved by intervening a CD47-beta amyloid protein signal axis through the CD47 monoclonal antibody, lung and kidney injuries are relieved, and the survival rate of the sepsis mouse is remarkably improved. Therefore, the CD47 can be used as a therapeutic target for preparation of drugs for treating sepsis. A monoclonal antibody / small molecule inhibitor / siRNA aiming at CD47 can also be used as a treatment strategy of sepsis.

The invention discloses a therapeutic chimeric monoclonal antibody and application thereof. The monoclonal antibody M01 is obtained from a mouse of an immune polypeptide vaccine MAP-Loop2, and the MAP-Loop2 polypeptide vaccine is a polypeptide fragment designed on the basis of a neisseria gonorrhoeae MtrE protein Loop2 structure. All sequences of immunized mouse B cells BCR are obtained through single cell sequencing, and a variable region sequence with the highest occurrence frequency is screened and cloned to a human IgG1 antibody constant region for expression and purification and is named as M01. Researches prove that the M01 has preventive and therapeutic effects on neisseria gonorrhoeae infection in a mouse vaginal infection model. According to the invention, the chimeric antibody with the Loop2 structure and exposed on the surface of the neisseria gonorrhoeae MtrE protein is prepared, so that a complement-dependent antibody sterilization reaction is effectively initiated; neisseria gonorrhoeae infection is effectively treated, the problem of drug resistance enhancement caused by antibiotic treatment is avoided, and a new treatment means is provided for gonorrhoeae prevention and control.

The invention discloses a multi-zone culture device for mouse cells, relates to the technical field of cell culture devices, and aims to solve the technical problem of low functionality of the multi-zone culture device for mouse cells, and the multi-zone culture device comprises a machine body, a base, a heating column, a rotating mechanism, a pulse mechanism, a plurality of culture components and an auxiliary mechanism. Through the structural design of the pulse mechanism, the bottom disc can rotate relative to the top disc, so that the sliding blocks can slide back and forth on the centripetal grooves, the net plate units synchronously move, the pressure of the culture cavity is increased and decreased, and the pulse type dynamic pressure in a mouse body is simulated; therefore, the accuracy and reliability of related research of mouse cell culture are improved, and the technical problem that a multi-zone culture device for mouse cells is low in functionality is solved.

The mammals rely on adaptive immunity to resist infections and tumors, wherein the thymus plays a critical role by generating T cells against infected or cancer cells. However, the function of the thymus declines along with the growth of age, and the immune system of the elderly is seriously damaged. For the study of reversing this decline, compositions and methods for activating and differentiating T cell progenitor cells in extrathymic tissue that stimulate the production of differentiated T cells are disclosed. For example, LNP-mediated delivery of mRNA encoding DLL-1, IL-7 and FLT3L to the liver has been demonstrated to create a temporary site for T cell maturation, enhancing T cell-mediated immunity in old mice without causing autoimmunity. According to the method, hematopoietic precursor cells are promoted to ectopic develop into T cells, dendritic cells are activated, and evidence that in-vivo engineering adaptive immunity can effectively relieve immune aging is provided.

The inventors have surprisingly demonstrated that GD3 positive senescent cells inhibit NK cell in vitro and in vivo while GD3 negative senescent cells is associated with NK cell functionality, both with human or murine cells. The inventors' results bring the proof of concept that GD3 expression may represent a Senescence-associated Immune Checkpoint (SIC) that determines senescent cell immunogenicity and identify GD3 and more generally SIC as a multi-hit target for age-associated diseases. Thus, GD3 may be a major step forward in the development of efficient anti-senescence immunotherapies. In particular, the present invention relates to a method for identifying whether a cell is in senescence process comprising the steps of: i) measuring the co-expression level of ST8Sia1 (GD3) with a senescence marker in a biological sample; ii) comparing the co-expression level measured at step i) with its predetermined reference value, and iii) concluding that the cell is in senescence process when the co-expression level of GD3 with a senescence marker is higher than its predetermined reference value or concluding that cell is not in senescence process when the co-expression level of GD3 with a senescence marker is lower or similar than its predetermined reference value. The present invention also relates a method for treating senescent cells accumulation related disease in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a GD3 inhibitor.