Dendritic cell inducer and preparation method and application thereof

A dendritic cell and inducer technology, applied in the field of nano-immune drugs, can solve the problems of loss of immunosuppressive function, low ability of body migration and homing, and impaired cell activity, etc., achieve adjustable surface physical and chemical properties, and increase clinical promotion And the possibility of application, the effect of low price

Active Publication Date: 2019-03-15
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are usually two ways to obtain tDCs, namely, the original imDCs directly or imDCs treated with traditional immunosuppressants, such as rapamycin (RAPA); ImDCs are induced to become mDCs and lose their immunosuppressive function, while imDCs treated with traditional immunosuppressants represented by RAPA often have impaired cell activity, and their in vivo migration and homing ability is still low, making it difficult to ensure sufficient cell migration to T lymphocyte-rich cells. Pools come into play

Method used

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  • Dendritic cell inducer and preparation method and application thereof
  • Dendritic cell inducer and preparation method and application thereof
  • Dendritic cell inducer and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1. Preparation of monolayer graphene oxide (graphene oxide, GO) with different sheet diameters

[0054] The graphene oxide used in the present invention can be prepared by referring to the prior art. It can be a two-dimensional nanomaterial with a sheet structure obtained by oxidation of graphene with a strong oxidant (such as potassium permanganate), or after the oxidation reaction of potassium permanganate in concentrated sulfuric acid and graphite powder, a brown in There are graphite flakes with derivatized carboxyl groups on the edge and mainly phenolic hydroxyl groups and epoxy groups on the plane. This graphite flake layer can be exfoliated into graphene oxide by ultrasonic or high-shear vigorous stirring, and forms a stable, light brown yellow in water. Monolayer graphene oxide suspension.

[0055] The following is a specific example to illustrate how to obtain graphene oxide with different sheet diameters:

[0056] 1g graphite flakes, 1g NaNO 3 , ...

Embodiment 2

[0065] Example 2. Culture and identification of immature dendritic cells derived from mouse bone marrow

[0066] 1. Culture and induction of immature dendritic cells derived from mouse bone marrow

[0067] Kill L2G85.C57BL / 6J mice (6-8 weeks) by cervical dislocation, soak them in 75% (v / v) alcohol and place them in an ultra-clean workbench to separate their femurs and tibias, soak them in PBS, and use aseptic Remove the muscle and epiphysis with scissors and tweezers, blow out the bone marrow in the bone marrow cavity with a 2mL syringe needle, absorb 1640 medium and wash the bone marrow cavity repeatedly until the bone marrow cavity turns white. Gently blow off the bone marrow cells with a syringe and filter through a 40 μm filter into a centrifuge tube.

[0068] Dispersed bone marrow cells were divided into 2 × 10 6 The cells / mL were cultured in a 6-well plate, and 1640 complete medium (containing 10 wt% fetal calf serum, 5 ng / mL IL-4, 10 ng / mL GM-CSF) was added to each we...

Embodiment 3

[0072] Example 3. Screening of neuropeptides

[0073] (1) Four neuropeptides inhibit DCs from secreting pro-inflammatory cytokines

[0074] The four neuropeptides UCN, α-MSH, VIP and AM were entrusted to Shanghai Gil Biochemical Company to process and produce them and were purified by high-performance liquid chromatography. HSDAVFTDNYTRLRKQMAVKKYLNSILN), cortistatin AM (YRQSMNQGSRSNGCRFGTCTFQKLAHQIYQLTDKDKDGMAPRNKISPQGY), melanocyte stimulating hormone releasing factor α-MSH (SYSMEHFRWGKPV).

[0075] Using UCN, α-MSH, VIP and AM four neuropeptides (respectively UCN-DCs group, VIP-DCs group, α-MSH-DCs group and AM-DCs group) to 10 -7 The immature dendritic cells were co-incubated at the final concentration of mol / L for 48 hours, and at the same time, a low dose of lipopolysaccharide (lipopolysaccharide, LPS) (200ng / ml) was given to stimulate the immature dendritic cells in a co-incubation manner, so as not to be treated with neuropeptides and given a low dose of LPS Stimulate...

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Abstract

The invention discloses a dendritic cell inducer and a preparation method and application thereof. The dendritic cell inducer is a compound of nervous peptide and graphene oxide, and dendritic cells can be induced into tolerant dendritic cells. Compared with other immunomodulators such as rapamycin and cocktail of cytokines, the compound is suitable for inducing the tolerant dendritic cells, has the advantages of simplicity in preparation, high stability and low price, and can be used for treating autoimmune diseases such as acute-graft-versus-host diseases, rheumatic arthritis and antoimmunediabetes.

Description

technical field [0001] The present invention relates to the technical field of nanoimmunopharmaceuticals, in particular to a dendritic cell inducer and its preparation method and application, in particular to an inducer for inducing immature dendritic cells into tolerant dendritic cells And its preparation method and application. Background technique [0002] Dendritic cells (Dendritic Cells, DCs) are the bridge connecting natural immunity and acquired immunity, and are the only type of Antigen Presenting Cells (Antigen Presenting Cells, APCs) that can activate naive T cells (Naive T Cells). Dendritic cells can mobilize a variety of immune resistance mechanisms, such as cytotoxic T cells, CD4 + Helper T cells, natural killer (NK) cells, and natural killer T (NKT) cells, etc. These dendritic lymphocytes all have the function of recognizing and killing diseased cells and releasing protective cytokines (such as IFN-γ, TNF-α). [0003] According to different cell phenotypes a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784A61K38/22A61P37/02A61K33/00A61K35/15
CPCA61K33/00A61K35/15A61K38/22A61P37/02C12N5/0639C12N2500/05C12N2501/30C12N2501/35C12N2501/855C12N2501/86A61K2300/00
Inventor 王小慧周欠欠詹林盛何楚琳
Owner ACADEMY OF MILITARY MEDICAL SCI
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