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A kind of m1 macrophage exosome vaccine and its preparation method and application

A technology of macrophages and exosomes, which is applied in the field of tumor immunotherapy drugs, can solve problems such as toxic side effects, high immunogenicity, and inability to effectively activate immune responses, so as to reduce toxic side effects, simple preparation method, and alleviate immunosuppression The effect of the phenomenon

Active Publication Date: 2022-06-28
HEBEI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to provide an M1 type macrophage exosome vaccine and its preparation method and application to solve the problem that only conventional immunotherapy methods cannot effectively activate the immune response in the immunosuppressive tumor microenvironment, and existing changes Drugs in the tumor microenvironment have problems such as high immunogenicity and potential side effects

Method used

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  • A kind of m1 macrophage exosome vaccine and its preparation method and application
  • A kind of m1 macrophage exosome vaccine and its preparation method and application
  • A kind of m1 macrophage exosome vaccine and its preparation method and application

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Embodiment 1

[0035] The following takes a widely used antigen model OVA as an example to introduce M1 in detail. OVA -Preparation method of Exos, such as figure 1 shown, including the following steps:

[0036] (1) Preparation of M1 macrophages carrying OVA

[0037] Female C57BL / 6 mice aged 6-10 weeks were selected, fasted the day before sacrifice, and starved for at least 8 h. After soaking and disinfecting with 75% alcohol, the operation is performed on the ultra-clean bench, and all items used must be clean and sterile. Fix the mouse to the foam board, lift the mouse abdominal fur with tweezers and cut open with scissors, do not cut the endothelium. Then lift the endothelium and cut a small mouth, hold it with tweezers, pour cold DMEM complete medium into the abdominal cavity, blow and suck a few times in the abdominal cavity, and gently stroke the abdomen of the mouse with fingers to fully lavage, collect the peritoneal lavage fluid, and repeat several times. times, centrifuged (100...

Embodiment 2

[0042] (1) M1 OVA -Exos in vitro immune activation ability evaluation

[0043] A large number of TAMs are present in the tumor microenvironment, and most of them exhibit an immunosuppressive M2 phenotype affected by the tumor microenvironment, which is a major obstacle to immunotherapy. We therefore adopted M1-Exos polarized macrophages as the M1 phenotype. like Figure 5 As shown, the results of the confocal laser show that M1-Exos and M1 OVA -Exos can increase the expression of CD80 and decrease the expression of CD206 on the surface of RAW264.7. This shows that M1-Exos is the same as M1 OVA -Exos can efficiently polarize macrophages to M1 phenotype. Image 6 Materials were analyzed for in vitro T cell activation capacity by flow cytometry. T cell Jurkat (clone E6-1) with PBS, OVA, M0-Exos, M1-Exos and M1 respectively OVA After co-incubation with -Exos, the cells were resuspended in PBS by centrifugation and CD4+ T cells and CD8+ T cells were labeled with fluorescent ...

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Abstract

The invention provides an M1 type macrophage exosome vaccine and its preparation method and application. The exosome vaccine is to allow M1 macrophages to ingest a specific tumor antigen Ag, and then extract the exosomes of M1 macrophages carrying the tumor antigen Ag to obtain the exosome vaccine M1 Ag -Exos. The present invention constructs an exosome vaccine that can regulate the tumor immune microenvironment to enhance the efficiency of immunotherapy, and utilizes M1 macrophage exosomes to realize the polarization of tumor-associated macrophages and the significant immune activation effect of tumor vaccines. The invention transforms the state of the macrophages from the state of immunosuppression promoting tumor invasion and metastasis to the state of immune support and anti-tumor by polarizing the tumor-associated macrophages into the M1 type. At the same time, through immune regulation of the tumor microenvironment, tumor vaccines can more efficiently promote the proliferation and activation of T cells, and effectively inhibit tumor growth and metastasis.

Description

technical field [0001] The invention relates to the technical field of tumor immunotherapy drugs, in particular to an M1 macrophage exosome vaccine and a preparation method and application thereof. Background technique [0002] Tumor immunotherapy has outstanding advantages and good therapeutic effects. However, in the process of tumor development, an immunosuppressive tumor microenvironment is gradually formed. TME is mainly composed of soluble factors such as blood vessels, immune cells, fibroblasts, extracellular matrix, and cytokine growth factors. The three main factors of poor immunogenicity of tumor cells, inhibition of effector T cell function, and a large number of immunosuppressive cells infiltrated in the tumor microenvironment cause the immunosuppression of TME, which seriously restricts the effect of immunotherapy. Macrophages are the main component of immunosuppressive cells and an important factor in immunosuppression. TAMs are classified into two functiona...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/00A61P35/00A61P37/02C12N5/0786
CPCA61K39/0011A61K39/001186A61P35/00A61P37/02C12N5/0645A61K2039/5154C12N2501/052C12N2501/998C12N2509/00
Inventor 李振华刘会芳吕芳芳张金超
Owner HEBEI UNIVERSITY
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