DC-based glioma holoantigen vaccine and preparation method thereof
A technology of glioma and whole tumor antigen, which is applied in the fields of bioengineering and biomedicine, can solve the problems of poor antigen stability, weak ability to stimulate T cells to secrete IFN-γ, and weak specificity, and achieve immunogenicity and Strong specificity, regulating the body's immune mechanism, and improving the effect of the body's immune mechanism
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[0023] A kind of DC-based glioma whole tumor antigen vaccine provided by the present invention, its preparation method comprises the following steps:
[0024] 1. Obtain immature DC;
[0025] 2. Add whole tumor antigen of glioma and co-culture with immature DC for 7-13 days;
[0026] 3. Add tumor necrosis factor-α, interleukin or lipopolysaccharide and continue culturing for 10-24 hours until DCs mature and DCs are loaded with glioma tumor antigens.
[0027] Specifically, in step 1, immature DCs can be derived from human blood, but since immature DCs only account for 0.5-1.0% of the total number of monocytes in blood, the number is very small; therefore, in a preferred embodiment of the present invention, adopt To obtain immature DCs by inducing monocytes, the specific steps are:
[0028] 11. Isolate monocytes from blood;
[0029] 12. Add in vitro induction medium and culture for 72 hours to amplify the monocytes obtained in step 11;
[0030] 13. Use the in vitro induction ...
Embodiment 1
[0064] The preparation method of the DC-based glioma whole tumor antigen vaccine comprises the following steps:
[0065] 1) Take 20ml of human peripheral blood, and use Ficon Lymphocyte Separation Medium to separate CD14 + Monocytes, after washing with PBS solution, press 2.5-5x10 7 / 3ml / well was inoculated in a six-well plate, and incubated with RPMI-1640 medium at 37°C, 5% CO 2 placed in an incubator for 90 minutes to collect adherent cells, namely CD14 + monocytes.
[0066] 2) CD14 + Monocytes were inoculated into RPMI-1640 medium containing 800 U / mL granulo-macrophage factor and 1000 U / mL interleukin-4 and incubated at 37°C, CO 2 After induction in an incubator with a concentration of 5% for 3-5 days, half of the medium was changed, and granule-macrophage factor and interleukin-4 were supplemented to maintain the concentration of granule-macrophage factor and interleukin-4 to obtain immature DC;
[0067] 3) Use PBS solution (PH=7.4) to adjust the concentration of imma...
Embodiment 2
[0075] The difference from Example 1 provided by the present invention is that in this example, the concentration of tumor necrosis factor-α is 10 ng / ml, the concentration of interleukin-1 is 5 ng / ml, and the concentration of glioma whole tumor antigen Concentration is 150ug / uL, immature DC concentration is 2×10 6 individual / mL.
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