人胞內(nèi)細(xì)胞因子檢測(cè)刺激方案
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| 人細(xì)胞因子 | 細(xì)胞來源 | 活化-極化培養(yǎng) | 培養(yǎng)時(shí)間 | 再次刺激 | 胞內(nèi)阻斷劑 |
| G-CSF | PBMC | LPS (1 ug/mL) | 24hr | - | Monensin |
| GM-CSF | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 5hr | - | Monensin |
| GIFN gamma | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 5hr | - | Brefeldin A |
| IL-1 alpha | PBMC | LPS (1 ug/mL) | 24hr | - | Monensin |
| IL-1 beta | PBMC | LPS (100ng/mL) | 4hr | - | Brefeldin A |
| IL-1RA | PBMC | LPS (100ng/mL) | 24hr | - | Brefeldin A |
| IL-2 | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 4-6hr | - | Brefeldin A |
| IL-4 | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 4-6hr | - | Brefeldin A |
| IL-5 | CD4 | Th2 polarizing cultures | 6d | PMA (50ng/mL) + Ionomycin (1ug/mL) (5hr) | Brefeldin A |
| IL-6 | PBMC | LPS (100ng/mL) | 24hr | - | Brefeldin A |
| IL-9 | CD4 | h2 polarizing culturesT | 6d | PMA (50ng/mL) + Ionomycin (1ug/mL) (5hr) | Monensin |
| IL-10 | CD4 | Th2 polarizing cultures | 6d | PMA (50ng/ml) + Ionomycin (1ug/ml) (5hr) | Monensin |
| IL-12/ IL-23 (p40) | PBMC |
hIFN gamma (100ng/mL) (2hr)/LPS (100ng/mL) (22hr) | 2hr/22hr | - | Brefeldin A |
| IL-13 | CD4 | anti-CD3 (10ug/mL, immobilized) + anti-CD28 (2ug/mL, soluble) + IL-2 (10ng/mL) + IL-4 (20ng/mL) (2d); IL-2 (10ng/mL) + IL-4 (20ng/mL) (3d) | 2d/3d | PMA (5ng/ml) + Ionomycin (500ng/ml) (4hr) | Brefeldin A |
| IL-17A | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 5hr | - | Monensin |
| IL-21 | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 4-7hr or 12-18hr | - | Brefeldin A |
| IL-22 | CD4 | Th17 polarizing cultures | 6d | PMA (50ng/ml) + Ionomycin (1ug/ml) (5hr) | Brefeldin A |
| IL-23 p19 | PBMC | hGM-CSF (40ng/mL) +hIL-4 (40ng/mL) | 6d | LPS (1ug/mL) (24hr) | Monensin |
| MCP-1/ CCL2 | PBMC | LPS (1ug/mL) | 24hr | - | Monensin |
| RANTES/ CCL5 | PBMC | LPS (1ug/mL) | 24hr | - | Monensin |
| TNF alpha | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 5hr | - | Brefeldin A |
| TNF beta | PBMC | Th1 polarizing cultures | 6d | PMA (50ng/mL) + Ionomycin (1ug/mL) (5hr) | Monensin |
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注： PBMC=外周血單核細(xì)胞peripheral blood mononuclear cell；PMA=佛波酯Phorbol Myristate Acetate; calcium ionophore; 鈣離子載體 iono=離子霉素ionomycin; LPS=脂多糖Lipopolysaccharide; 4hr=4 hours culture; 2d=2 days culture. GolgiStop/GolgiPlug: 一種胞內(nèi)阻斷劑，可以用BFA/Monensin替代。
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