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A mesenchymal stem cell that blocks th17 signaling pathway for treating autoimmune diseases, its preparation method and application

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: 2021-02-19
北京贝来生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, clinical studies have also found that a small number of patients have serious adverse events when treating patients with the above monoclonal antibodies, including coronary artery disease, congestive heart failure, viral syndrome, urinary tract infection, surgical wound cellulitis and elevated liver enzyme levels , this adverse reaction is more common in patients with hyperglycemia, hypertension and hyperlipidemia[14]
In addition, anti-mAb antibodies were detected in the serum of some patients (4%) during the use of the above-mentioned antibodies [14], suggesting that resistance to targeted drugs is still a problem that cannot be overcome by monoclonal antibody drugs

Method used

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  • A mesenchymal stem cell that blocks th17 signaling pathway for treating autoimmune diseases, its preparation method and application
  • A mesenchymal stem cell that blocks th17 signaling pathway for treating autoimmune diseases, its preparation method and application
  • A mesenchymal stem cell that blocks th17 signaling pathway for treating autoimmune diseases, its preparation method and application

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 using the fourth-generation lentiviral vector system to construct the αIL23:Fc fusion protein gene lentiviral expression plasmid, 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 cells after virus transfection was detected by 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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PUM

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Abstract

The invention discloses a mesenchymal stem cell for treating autoimmune diseases by blocking the Th17 signaling pathway. The mesenchymal stem cell highly expresses the αIL23:Fc fusion protein of the anti-IL-23 single-chain antibody and the Fc fragment of IgG1 by means of genetic modification . The αIL23:Fc fusion protein can competitively inhibit the binding of IL-23 to its receptor, so that it can be used to inhibit IL-23-mediated inflammation-related signaling pathways and inflammatory responses. The invention also discloses the preparation method and application of the mesenchymal stem cells.

Description

technical field [0001] The invention relates to a mesenchymal stem cell for blocking Th17 signaling pathway for treating autoimmune diseases, a preparation method and application thereof, and belongs to the field of biomedicine. Background technique [0002] Human interleukin 23 (hIL-23) is secreted by dendritic cells (DC cells), macrophages or other antigen-presenting cells, and is a heterodimer composed of two subunits of p19 and p40 linked by disulfide bonds Structural inflammatory factor [1]. IL-23 and IL-12 share the p40 subunit, and its coding gene is located on human chromosome 5; while p19 is a unique subunit protein of IL23, and its coding gene is located on human chromosome 12. Research and development found that [2], IL23 mediates many inflammatory responses in the body by binding to its receptor (IL-23R). [0003] IL-23R is also a heterodimer structure, consisting of the IL-12Rβ1 subunit shared with IL-12R (which combines with the IL-12p40 subunit), and its own...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/62C12N15/867A61K35/51A61P37/02A61P19/02A61P17/06A61P29/00A61P11/00A61P1/00
CPCA61K35/51A61P1/00A61P11/00A61P17/06A61P19/02A61P29/00A61P37/02C07K16/244C07K2319/02C07K2319/30C12N15/86C12N2510/00C12N2740/15043
Inventor 刘广洋刘拥军
Owner 北京贝来生物科技有限公司
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