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A whole-transcriptome expression profile of exosomes and its construction method and application

A construction method and exosome technology, applied in the field of transcriptome, can solve the problem that basophils have not been activated to release exosomes

Active Publication Date: 2021-05-25
AFFILIATED HOSPITAL OF GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the release of exosomes from the activation of basophils

Method used

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  • A whole-transcriptome expression profile of exosomes and its construction method and application
  • A whole-transcriptome expression profile of exosomes and its construction method and application
  • A whole-transcriptome expression profile of exosomes and its construction method and application

Examples

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Embodiment 1

[0037] An embodiment of the method for constructing the whole transcriptome expression profile of exosomes released by basophils in the present invention, comprising the steps of: preparation, activation and cultivation of human peripheral blood basophils, collecting cell cultured Supernatant, preparation of exosomes, extraction of total RNA from exosomes, whole transcriptome sequencing of exosomes, and analysis of changes in the expression profile of the whole transcriptome.

[0038] The groups of exosomes released before the activation of human basophils provided in this example are Control 1, Control2, and Control 3, and the corresponding groups of exosomes released after activation of human basophils are Anti- IgE 1, Anti-IgE 2, Anti-IgE 3.

[0039] Among them, human peripheral blood Preparation, activation and culture of basophils , including the following steps:

[0040] (1) Specimen collection: After obtaining the informed consent of healthy volunteers, collect EDTA a...

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Abstract

The invention discloses a method for preparing exosomes released by activated basophils, comprising the steps of: preparing basophils from anticoagulated peripheral blood of normal people by negative selection, passing Anti-IgE To stimulate activation, use serum-free culture at 37°C with 5% CO 2 Cultivate in an incubator for 12 to 48 hours, then collect the supernatant of the cell culture solution, centrifuge at room temperature at 300g, collect the supernatant of the culture solution after centrifugation, and obtain it by ultracentrifugation; the invention also discloses the full transcription of the above-mentioned exosomes Group expression profiles and methods for their construction. The present invention successfully isolated and identified the exosomes released by human basophils for the first time, and established the whole transcriptome expression profile of exosomes released by human basophils for the first time, and identified multiple post-activation differences The expressed RNA provides a new target for further research on the molecular mechanism of the interaction between activated basophils and target cells in the occurrence of diseases.

Description

technical field [0001] The present invention relates to the technical field of transcriptome, in particular to a whole-transcriptome expression profile of exosomes released by activated basophils, its construction method and application. Background technique [0002] Basophil (Basophil, Ba) is a small group of peripheral blood leukocytes, and is the main effector cell of allergic diseases. In recent years, its function in immune regulation has attracted great attention of researchers. Studies have found that basophils are abnormally activated in patients with rheumatoid arthritis and systemic lupus erythematosus and in mouse models and are related to disease activity; activated basophils can participate in the development of RA by affecting the balance of Th1 / Th2 Development; it can promote the differentiation and maturation of B cells into plasma cells and synergize with IL-3 to enhance antibody production. A variety of pathways can mediate the activation of basophils, bu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0787C12Q1/6811A61K45/00A61P37/08A61P37/02
CPCA61K45/00A61P37/02A61P37/08C12N5/0642C12N2509/00C12Q1/6811C12Q2535/122C12Q2539/103
Inventor 潘庆军刘华锋廖淑珍招春飞陈小群陆兴杨陈
Owner AFFILIATED HOSPITAL OF GUANGDONG MEDICAL UNIV
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