DC cell preparation method for improving antigen presenting T cell and improving T cell killing efficiency and application thereof
A cell and antigen technology, applied in the field of preparation of DC cells, can solve the problems of low antigen presentation efficiency, unsatisfactory T cell-specific immune response effect, and inability to exert anti-tumor effect, so as to achieve high antigen presentation efficiency and improve killing effect. Efficiency, activation-promoting effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0040] Example 1 Preparation method of dendritic cells that can improve antigen-presenting T cells and increase T-cell killing activity
[0041] 1. Preparation of DC cells
[0042] 1.1 Add 5ml of peripheral blood serum into a T75 bottle, and cover in the dark at 37°C for 4 hours.
[0043] 1.2 Pour fresh heparin-anticoagulated human peripheral blood into a centrifuge tube, balance, and centrifuge at 700g / min for 20 minutes (the slowest rate of decline).
[0044] 1.3 Add 1:1 D-PBS to the cell layer after centrifugation of the above-mentioned peripheral blood and mix well, slowly add the lymphocyte separation liquid along the tube wall at a ratio of 1:1 to the surface of the lymphocyte separation liquid, and keep a clear interface. After centrifugation at 800g for 15min (slow rise and fall), the centrifuge tube is divided into four layers from top to bottom.
[0045] Among them, the first layer is a D-PBS layer, the second layer is a ring-shaped milky white PBMC, the third laye...
experiment example 1
[0142] Experimental Example 1 Flow cytometric detection of DC maturation ratio
[0143] The DC cell surface markers HLA-DR+, CD1a+, CD83+ obtained in the embodiment and 4 groups of control examples were detected by flow cytometry, and the detection results were as follows: figure 1 shown. Depend on figure 1 From the results, it can be seen that the expression levels of markers on the surface of DC cells obtained in the examples were higher than those in the 4 groups of control examples. It can be concluded that the rate of obtaining mature DC in the example is higher than that in the 4 groups of control examples.
experiment example 2
[0144] Experimental Example 2 Detection of T cells and their secretion of cytokines
[0145] 1) Use flow cytometry to detect the maturation ratio of DC-stimulated T cells obtained in the example and the 4 groups of control examples; the results are as follows Figure 2-6 shown. The results showed that the ratio of DC cells obtained in the embodiment to stimulate T cell maturation was 91.78% ( figure 2 ), the DC cells obtained in Control Example 1 stimulated T cell maturation ratio was 82.31% ( image 3 ), the ratio of DC cells obtained in Control Example 2 to stimulate T cell maturation was 74.96% ( Figure 4 ), the DC cells obtained in Control Example 3 stimulated T cell maturation ratio was 78.24% ( Figure 5 ), the ratio of DC cells obtained in Control Example 4 to stimulate T cell maturation was 62.31% ( Figure 6 ) suggest that DCs obtained by the method described in this application can more effectively activate antigen-presenting T cells.
[0146] 2) ELISA detecti...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com