M1 type macrophage exosome vaccine as well as preparation method and application thereof

A macrophage and exosome technology, applied in the field of tumor immunotherapy drugs, can solve the problems of ineffective activation of immune response, high immunogenicity, toxic and side effects, etc., and achieves alleviation of immunosuppressive phenomenon, simple preparation method, and reduced toxicity. Effects of side effects

Active Publication Date: 2021-09-17
HEBEI UNIVERSITY
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  • 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

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  • M1 type macrophage exosome vaccine as well as preparation method and application thereof
  • M1 type macrophage exosome vaccine as well as preparation method and application thereof
  • M1 type macrophage exosome vaccine as well as preparation method and application thereof

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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 and fasted for at least 8 h the day before sacrifice. After soaking and disinfecting with 75% alcohol, operate 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 forceps, and cut it with scissors, do not cut the inner skin. Then lift the endothelium and cut a small opening, 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 touch the abdomen of the mouse with your fingers to fully lavage, collect the peritoneal lavage fluid, repeat several times Centrifuge (1000 rpm, 5 min), resuspend with DMEM com...

Embodiment 2

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

[0043] There are a large number of TAMs in the tumor microenvironment, and affected by the tumor microenvironment, most of them show an immunosuppressive M2 phenotype, which is a major obstacle for immunotherapy. So we adopted M1-Exos polarized macrophages as the M1 phenotype. like Figure 5 As shown, the results of 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 and M1 OVA -Exos can efficiently polarize macrophages to the M1 phenotype. Image 6 The in vitro T cell activation ability of the material was analyzed by flow cytometry. T cell Jurkat (clone E6-1) with PBS, OVA, M0-Exos, M1-Exos and M1 OVA - After co-incubation with Exos, centrifuge PBS to resuspend the cells and label CD4+T cells and CD8+T cells with fluorescent antibodies, respectively. Detected by flow cytom...

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Abstract

The invention provides an M1 type macrophage exosome vaccine as well as a preparation method and application thereof. The M1 type macrophage exosome vaccine is characterized in that an exosome vaccine M1Ag-Exos is obtained by enabling M1 type macrophages to uptake a specific tumor antigen Ag and then extracting an exosome of the M1 type macrophages carrying the tumor antigen Ag. According to the M1 type macrophage exosome vaccine, the exosome vaccine capable of adjusting the tumor immune microenvironment is constructed to enhance the immunotherapy efficiency, and polarization of tumor-related macrophages and the remarkable immune activation effect of a tumor vaccine are achieved through the M1 type macrophage exosome; the tumor-related macrophages are polarized into M1 type, so that the macrophages are converted from a state of promoting tumor invasion and metastasis by immunosuppression into a state of supporting tumors by immunity; meanwhile, through immunoregulation on the tumor microenvironment, the tumor vaccine can promote proliferation and activation of T cells more efficiently, and growth and metastasis of the tumors are effectively inhibited.

Description

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

Claims

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Application Information

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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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