Antigen-specific immature DC (dendritic cell) source exosome as well as preparation and application methods thereof

An application method and specific technology, applied in the direction of medical materials derived from mammals, animal cells, neuromuscular system diseases, etc., can solve the problem of non-antigen-specific, antigen-specific treatment of autoimmune diseases, and inability to achieve treatment Effect and other issues

Inactive Publication Date: 2014-08-06
XIANGYA HOSPITAL CENT SOUTH UNIV
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Problems solved by technology

However, the immature DC used in these experiments is not antigen-specific, so it cannot produce antigen-specific therapy for autoimmune diseases and cannot achieve the best therapeutic effect

Method used

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  • Antigen-specific immature DC (dendritic cell) source exosome as well as preparation and application methods thereof
  • Antigen-specific immature DC (dendritic cell) source exosome as well as preparation and application methods thereof
  • Antigen-specific immature DC (dendritic cell) source exosome as well as preparation and application methods thereof

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Embodiment Construction

[0037] The following in conjunction with specific embodiments is intended to further illustrate the present invention, rather than limit the present invention.

[0038] 1. Preparation of antigen-specific immature DC-derived exosomes

[0039] 1.1 Isolation and purification of myeloid-derived immature DC and phenotypic detection

[0040] Select 6-8 week old B6 mice, take the femur and tibia under sterile conditions, lyse the bone marrow fluid of the tibia and fibula with red blood cells to remove red blood cells, then add RPMI1640 complete medium containing 10ng / ml rmGM-CSF to culture for 72 hours and then the full amount The culture was continued until the fifth day after changing the medium, and then cultured until the seventh day after changing the medium in half. According to the instructions of magnetic bead sorting, select anti-CD11c magnetic beads to sort and purify DC, so that the purity can reach more than 95% (attached figure 1 ).

[0041] 1.2 Flow cytometry detecti...

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Abstract

The invention discloses an antigen-specific immature DC (dendritic cell) source exosome as well as preparation and application methods thereof. The maturity of a bone marrow derived dendritic cell (BMDC) is interfered by using microRNA-146a mimics, and then through detecting the expression of CD80, CD86 and MHC II molecules on the surface of the BMDC, a situation that the BMDC transfected by the microRNA-146a mimics has a capacity of resisting antigen mature stimulation is confirmed; and the transfected BMDC loads the pathogenic peptide fragment of MG, so that an antigen-specific immature DC is obtained, and an exosome secreted by the antigen-specific immature DC is the exosome provided by the invention. The exosome is used for treating myasthenia gravis, and can be further used for preparing medicines or preparations for treating myasthenia gravis; and the exosome has the advantages of simple preparation method, easiness for storage, strong stability, remarkable effect, strong specificity, small side effects, and the like, therefore, the exosome has a broad application prospect.

Description

[0001] Invention name [0002] The invention belongs to the technical field of drugs or preparations for treating myasthenia gravis, and specifically relates to an antigen-specific immature DC-derived exosome and a preparation method thereof, and its application in the preparation of drugs for treating myasthenia gravis. Background technique [0003] Myasthenia Graves (MG) is a common autoimmune disease of the nervous system. The incidence rate is (0.5-5) / 100,000, and the prevalence rate is 10 / 100,000, and it is still on the rise. The clinical manifestations are fluctuations in the skeletal muscles of the whole body and fatigue, and the symptoms are mild in the morning and severe in the evening. In severe cases, the respiratory muscles may be involved and life-threatening. The disease is easy to relapse and difficult to cure, which significantly affects the quality of life and survival of patients. Traditional treatment methods include thymectomy, plasma exchange, and applic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784A61K35/14A61P21/04
Inventor 杨欢尹炜凡
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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