New application of isoastragaloside II in preparation of tumor immune treatment drugs
A technology of isoastragaloside and astragaloside, which is applied in the field of isoastragaloside II in the preparation of tumor immunotherapy drugs, and can solve problems such as insufficient research
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Embodiment 1
[0035] Example 1: Isastragaloside II enhances CD8 + T cell immune response
[0036] Experimental materials: 8-week-old male C57 mice (purchased from Victoria Lihua Company), anti-mouse CD3, anti-mouse CD28 (BD), IL-2 (R&D), PE-anti-mouse CD8a; APC-anti- mouse IFN-γ; Per CP anti-mouseperforin (BD), CFSE dye, propylene glycol methyl ether acetate (PMA); ionomycin (purchased from sigma), fetal bovine serum, 1640 medium, DMEM culture base (Gibco), isoastragaloside Ⅱ (sigma);
[0037] Test content:
[0038] 1. Preparation of spleen single-cell suspension: kill the mouse, take the spleen, crush the tissue with a 200-mesh filter, collect the cells in 1640 culture medium, centrifuge at 1400 r / min to discard the supernatant, use the erythrocyte lysate to remove the red blood cells, and collect the lysate Then add 10% fetal bovine serum 1640 culture medium to the cells for later use;
[0039] 2. Use anti-mouse CD3 (10μg / mL) to coat the plate overnight;
[0040] 3. Use anti-mouse CD...
Embodiment 2
[0046] Example 2: Effect of isoastragalosides on the killing effect of CTL cells
[0047] Experimental materials: 8-week-old male C57 mice (purchased from Victoria Lihua Company), anti-mouse CD3; anti-mouse CD28 (BD), IL-2 (R&D), PE-anti-mouse CD8a; APC-anti- mouse IFN-γ; PerCP anti-mouseperforin (BD), CFSE dye; propylene glycol methyl ether acetate (PMA); ionomycin (purchased from sigma), fetal bovine serum; 1640 medium, DMEM medium (Gibco), isoastragaloside II (sigma);
[0048] Test content
[0049] 1. Preparation of spleen single-cell suspension: kill the mouse, take the spleen, crush the tissue with a 200-mesh filter, collect the cells in 1640 culture medium, centrifuge at 1400 r / min to discard the supernatant, use red blood cell lysate to remove red blood cells, and collect and lyse Afterwards, the cells were added with 10% fetal bovine serum 1640 culture medium for later use.
[0050] 2. Use anti-mouse CD3 (10μg / mL) to coat the plate overnight
[0051] 3. Use anti-mo...
Embodiment 3
[0057] Example 3: Isastragaloside II regulates the killing effect of CTL
[0058] Experimental materials: 8-week-old male C57 mice (purchased from Victoria Lihua Company), anti-mouse CD3; anti-mouse CD28 (BD), IL-2 (R&D), PE-anti-mouse CD 8a; APC-anti -mouse IFN-γ; PerCP anti-mouseperforin (BD), CFSE dye; propylene glycol methyl ether acetate (PMA); ionomycin (purchased from sigma), fetal bovine serum; 1640 medium; DMEM culture Gibco (Gibco), Isastragaloside II (sigma)
[0059] Test content
[0060] 1. Preparation of spleen single-cell suspension: kill the mouse, take the spleen, crush the tissue with a 200-mesh filter, collect the cells in 1640 culture medium, centrifuge at 1400 r / min to discard the supernatant, use red blood cell lysate to remove red blood cells, and collect and lyse Then add 10% fetal bovine serum 1640 culture medium to the cells for later use;
[0061] 2. Use anti-mouse CD3 (10μg / mL) to coat the plate overnight;
[0062] 3. Use anti-mouse CD28 (1μg / mL)...
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