Preparation method and application of tolerogenic dendritic cells capable of keeping stability in inflammatory environment

Abstract

The invention relates to a preparation method and application of tolerogenic dendritic cells capable of keeping stability in the inflammatory environment. The preparation method is characterized by using a monomer structure compound for carrying out targeted inhibition on phosphorylation of mTOR signal channel protein, and carrying out in-vitro tolerance short-time induction and culture of human dendritic cells. The result shows that the prepared tDC still maintains a stable tolerance regulation function when the tDC is irritated by inflammatory molecules. A new tDC preparation method is builtand is capable of ensuring the survival rate of cell products in moderate concentration and time and preparing tDC cell products which can still stably maintain the tolerance function in the inflammatory environment; the raw materials adopted by the preparation method easily pass GMP authentication; the preparation process is simple, convenient and economical; the prepared products are stable andeffective and are easily standardized; the prepared tDC can be applied to adoptive infusion, can be used for treating multiple allergic immune reactive diseases, and has great potential in clinical application.

Description

technical field [0001] The invention relates to the technical field of in vitro induction and culture of cells, in particular to a preparation method and application of stable tolerance dendritic cells in an inflammatory environment. Background technique [0002] The body's immune system plays an important role in eliminating "non-self" and maintaining a stable internal environment. Once the immune system is abnormally enhanced and triggered by exogenous or endogenous factors, allergic immune response diseases will occur, mainly including Allergic diseases (allergic asthma, allergic purpura, etc.), autoimmune reactivity (multiple sclerosis, myasthenia gravis, etc.) and graft-versus-host disease (transfusion-related immune reactions, bone marrow and organ transplantation-related GVHD). The lighter ones are protracted, and the severe ones are life-threatening. These patients' own immune cells are often in a state of severe dysregulation, so adoptive tolerance cells are induce...

AI Technical Summary

Problems solved by technology

The use of TGF-β and / or IL-10 induction methods has the following disadvantages: 1) The cytokines are not easy to store, the titer is unstable, and the stability of the induced tDC cannot be guaranteed, and the preparation method cannot guarantee GMP certified products Possibility of repeatability; 2) Commercially available cytokines mainly rely on expression and purification of bacterial recombinant proteins, and most of them are limited to experimental use, which is also a bottleneck that hinders the clinical application of immune cell products in the future; 3) DC is a kind of highly differentiated The required cell products, in order to ensure the high stability of the cell products, usually require extremely high purity for the external stimuli and signals received during the derivation process to ensure that they are not induced into other cell types, that is, for TGF-beta and IL-10 The purity requirements are very high, which leads to an increase in the cost of preparation, which also increases the cost of therapeutic cells for industrial use; 4) The preparation of tDC by cytokine induction often requires 3-7 days of DC treatment, and the induction process is time-consuming Labor consumption increases the instability of cell products and the difficulty of standardization, which will increase the difficulty of GMP certification in the future; 5) The internal environment of clinical patients is usually complicated, and there are often multiple pathogen infections due to surgery and hospital environments. How tolerogenic cells can maintain a high level of tolerance and regulatory functionality in the complex infectious and inflammatory environment infused into the body is the key to the clinical value of this cell therapy

However, in the prior art, there is no report on the tolerance dendritic cells and their preparation methods that are stable in the inflammatory environment of the present invention.

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