Whole-transcriptome expression profile of exosome, and construction method and application of whole-transcriptome expression profile

Abstract

The invention discloses a preparation method of an exosome released by activated basophil. The preparation method comprises the steps of preparing basophil from anticoagulant peripheral blood of a normal person by a negative selection method, performing activation through Anti-IgE stimulation, performing culture in an incubator containing 5% of CO2 at 37 DEG C through serum-free culture mediums for 12-48 hours, then collecting supernatant of cell culture fluid, performing centrifuging under the condition of 300g at room temperature, collecting culture fluid supernatant after the centrifuging,and performing ultracentrifugation so as to obtain the exosome released by activated basophil. The invention also discloses a whole-transcriptome expression profile of the exosome, and a constructionmethod of the whole-transcriptome expression profile. According to the preparation method disclosed by the invention, the exosome released by human basophil is successfully separated and identified for the first time, besides, the whole-transcriptome expression profile of the exosome released by the human basophil is built for the first time, a plurality of RNAs having difference expression afteractivation are identified, and a new target point is provided for further researching a molecular mechanism that the activated basophil acts with target cells during disease generation.

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...

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