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Mesenchymal stem cell capable of blocking Th17 signaling pathway to treat autoimmune diseases, and preparation method and application thereof

A technology for autoimmune diseases and stem cells, applied in genetically modified cells, botanical equipment and methods, biochemical equipment and methods, etc.

Active Publication Date: 2019-03-29
北京贝来生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

This technology can be useful for various purposes such as improving efficiency or reducing costs associated with certain processes.

Problems solved by technology

This patents describes how certain types of molecules called cytokines play important roles in regulation of various biological processes like inflammehnogenesis, neutrophylaxis, nerve growth factor induction, platelet aggregability, astrocytotic activity, apoptosis, neurotransmitter functions involved in neuron function expression, migration into brain areas related to their involvement in multiple sclerosis symptoms, systematic organ damage caused by excessively active TH 17 cells associated with chronic collagenopathy, dermal fibromatoses, skin ulcers, vasculitis, pulmonemia, glomeruloscidemneys, pyoderma ganglia, sarcoidosis, hemophilia A, thrombohemolytic microangioprolites, uvexanthema, alvacrophage activation, lung injury, sepsis, acute phase reactant cycle virus induced conditions, cardiovascular complications, renal impairments, cancer metastasis, graft versus host disease, infectious agent shocks, and others.

Method used

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  • Mesenchymal stem cell capable of blocking Th17 signaling pathway to treat autoimmune diseases, and preparation method and application thereof
  • Mesenchymal stem cell capable of blocking Th17 signaling pathway to treat autoimmune diseases, and preparation method and application thereof
  • Mesenchymal stem cell capable of blocking Th17 signaling pathway to treat autoimmune diseases, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: Construction of LV-αIL23: Fc fusion protein gene vector

[0047] The αIL23:Fc gene plasmid we designed was constructed by using the fourth-generation lentiviral vector system to construct the lentiviral expression plasmid of αIL23:Fc fusion protein gene, and LV-eGFP was used as the control virus. ( figure 1 For LV-αIL23:Fc fusion protein and LV-eGFP plasmid construct)

Embodiment 2

[0048] Example 2: LV-αIL23: Fc fusion protein genetically modified mesenchymal stem cells

[0049] Mix the above-mentioned LV-αIL23:Fc fusion protein particles and LV-eGFP with lentiviral pGag / Pol, pRev, pVSV-G and other framework plasmids respectively, introduce them into 293T cells through LTX liposomes, and package mature lentiviruses. After the viruses are harvested MSCs were transfected, and puromycin was added 24 hours later to select successfully transfected cells.

Embodiment 3

[0050] Example 3: Immunofluorescence detection of gene-modified stem cells expressing green fluorescent protein

[0051] The green fluorescence of the cells after virus transfection was detected by a fluorescence microscope, and it was found that both LV-eGFP and LV-αIL23:Fc fusion proteins could successfully transfect MSCs cells, and the transfection rate reached more than 90% ( figure 2 Fluorescence microscopy was used to detect the transfection effect of gene-modified MSCs).

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Abstract

The invention discloses a mesenchymal stem cell capable of blocking a Th17 signaling pathway to treat autoimmune diseases. The mesenchymal stem cell highly expresses an [alpha]IL23:Fc fusion protein of an anti-IL-23 single-chain antibody and an IgG1 Fc fragment by gene modification. The [alpha]IL23:Fc fusion protein can competitively inhibit the binding of IL-23 and an acceptor, so as to be used for inhibiting IL-23-mediated inflammation-related signaling pathways and inflammatory response. The invention also discloses a preparation method and application of the mesenchymal stem cell.

Description

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Claims

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Application Information

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Owner 北京贝来生物科技有限公司
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