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Exosome-based anti-tumor vaccine

An exosome and tumor technology, applied in the field of exosomes, can solve the problems of unsatisfactory tumor inhibition effect and complex tumor microenvironment.

Active Publication Date: 2020-02-28
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, many new nano-tumor vaccines have been proved in in vitro experiments to achieve the above-mentioned cellular immune response and generate a large number of CTLs, but the tumor-suppressing effect at the animal level is not ideal, mainly due to the existence of complex tumors at the tumor site in animals. Microenvironment

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  • Exosome-based anti-tumor vaccine
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Examples

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preparation example Construction

[0052] The present invention also provides a method for preparing exosomes, which includes the following steps: extracting the nucleus of tumor cells, adding the tumor cell nucleus to the culture medium of antigen-presenting cells, and collecting the supernatant of the culture medium after incubation. centrifuge to collect exosomes.

[0053] In a preferred embodiment of the present invention, the antigen-presenting cells phagocytize tumor cell nuclei to form hybrid cells, and the hybrid cells secrete exosomes, and the exosomes are a type of hybrid exosomes.

[0054] In a preferred embodiment of the present invention, the antigen-presenting cells are dendritic cells, macrophages or B cells, more preferably the antigen-presenting cells are macrophages.

[0055] In a preferred embodiment of the present invention, the endogenous expression of tumor antigens on macrophages is successfully achieved through the strategy of macrophages engulfing tumor cell nuclei. In addition, tumor ...

Embodiment 1

[0073] Example 1, Preparation of E.G7-macrophage hybrid cells

[0074] (a) Cell nuclei derived from lymphoma cell E.G7-OVA (purchased from ATCC cell bank) were extracted using a nucleus extraction reagent (purchased from Solarbio):

[0075] E.G7-OVA cells were cultured in a medium containing 10% fetal bovine serum albumin (purchased from Gibco), 1% penicillin-streptomycin (purchased from Gibco), 0.4mg / mLG418 (purchased from Gibco) and 0.05mMβ- Mercaptoethanol (purchased from Gibco) in high glucose 1640 medium (purchased from Gibco). Subculture once every two days at a ratio of 1:5. The tumor cells in the logarithmic growth phase were collected by centrifugation, resuspended in PBS (Gibco Company), and washed twice. The cell pellet was resuspended using the Lysis buffer in the cell nucleus extraction kit (purchased from Solarbio). After resuspending, the cell density was about 2×10 7 / mL. Afterwards, 20 μL of ReagentA (purchased from Solarbio) was added to each milliliter o...

Embodiment 2、B16

[0079] Example 2, Preparation of B16-macrophage hybrid cells

[0080] (a) Cell nuclei derived from melanoma cells Muc-B16 (purchased from ATCC cell bank) were extracted using a nucleus extraction reagent (purchased from Solarbio):

[0081] Muc1-B16 cells were cultured on the wall, and the complete medium contained fetal bovine serum albumin (purchased from Gibco), 1% penicillin-streptomycin (purchased from Gibco), 1 μg / mL puromycin (purchased from Gibco) 1640 medium (purchased from Gibco, without HEPES), and passaged once every 2-3 days according to the ratio of 1:4. The cells in the logarithmic phase were collected by centrifugation, resuspended in PBS (purchased from Gibco), and washed twice. Will 2×10 7 Each cell was resuspended in Lysis buffer in 1 mL cell nucleus extraction kit (purchased from Solarbio Company), and then 20 μL ReagentA (purchased from Solarbio Company) was added to each milliliter of cell suspension, blown 10 times with a 1 mL pipette tip, and waited W...

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Abstract

The invention relates to an exosome, which is a hybrid cell secretion formed by ingesting tumor nuclei by antigen presenting cells. Through a strategy where tumor nuclei are ingested by macrophages, endogenous expression of tumor antigens on the macrophages is achieved; the exosome prepared has good lymph node and tumor dual-targeting capability.

Description

technical field [0001] The present invention relates to an exosome, its preparation method and an anti-tumor immunotherapy method based on the exosome. The exosome has dual effects of immune activation and tumor microenvironment regulation. [0002] technical background [0003] There are usually three types of treatment for malignant tumors: surgery, radiation therapy, and chemotherapy. Among them, surgical treatment is limited to the direct resection of solid tumors visible to the naked eye to achieve therapeutic effects, and the malignant tumors are extremely infiltrating. Surgical treatment cannot guarantee the removal of all tumor cells, and there is a high risk of tumor recurrence. Radiation therapy is the use of radiation with cell-killing ability (such as: α-rays, β-rays, γ-rays, etc.) to irradiate malignant tumors, thereby killing cancer cells. Since ray radiation therapy is not specific, it will inevitably have certain adverse effects on surrounding tissues and cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/16A61K39/00A61P35/00
CPCC12N5/16C12N5/163C12N5/12A61K39/0011A61P35/00A61K2039/5154A61K45/06C12N5/10C12N5/0645A61K35/15A61K35/17A61K35/13A61K2300/00A61K2039/572C07K16/2818C07K16/2827
Inventor 马光辉魏炜王双卿爽王江华
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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