小鼠胞内细胞因子检测刺激方案
| 小鼠细胞因子 | 细胞来源 | 活化-极化培养 | 培养时间 | 再次刺激 | 胞内阻断剂 |
| GM-CSF | mouse spleen | ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d) | 2d/3d | anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr | Brefeldin A |
| IFN-gamma | mouse spleen | ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d) | 2d/3d | anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr | Brefeldin A |
| IL-1 alpha | mouse PEC | mIFNγ (100ng/mL) (2hr)/LPS (100ng/mL)(22hr) | 2hr/22hr | - | Brefeldin A |
| IL-1 beta | mouse PEC | LPS (100 ng/mL) (22hr) | 22hr | - | Monensin |
| IL-2 | mouse spleen | ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d) | 2d/3d | anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr | Brefeldin A |
| IL-4 | mouse spleen | Th2 polarized | 6d | PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr | Brefeldin A |
| IL-5 | mouse splenic CD4 | ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d) | 2d/3d | immobilized) + anti-CD28 (2ug/mL soluble) 5hr | Brefeldin A |
| IL-6 | mouse PEC | LPS (100 ng/mL) (22hr) | 22hr | - | Monensin |
| IL-10 | mouse spleen | ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d) | 2d/3d | anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr | Brefeldin A |
| IL-12/IL-23 (p40) | mouse PEC | LPS (100 ng/mL) (22hr) | 22hr | - | Brefeldin A |
| IL-13 | mouse spleen | Th2 polarized | 6d | PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr | Brefeldin A |
| IL-17A | mouse spleen | Th17 polarized | 6d | PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr | Monensin |
| IL-17F | mouse spleen | Th17 polarized | 6d | PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr | Monensin |
| IL-21 | mouse spleen | Th17 polarized | 9d | PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr | Monensin |
| IL-22 | mouse spleen | Th17 polarized | 12d | PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr | Brefeldin A |
| IL-23 p19 | mouse bone marrow | mGM-CSF (40 ng/mL) | 8d | LPS (1 ug/mL) (24 hr) | Monensin |
| MCP-1/ CCL2 | mouse PEC | LPS (100 ng/mL) (24hr) | 24hr | - | Brefeldin A |
| TNF alpha | mouse spleen | ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d) | 2d/3d | anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr | Brefeldin A |
| 说明：mouse PEC=mouse thioglycolate-elicited peritoneal macrophages腹腔巨噬細胞; ConA=刀豆素Concanavalin A; Iono=离子霉素Ionomycin; LPS=脂多糖Lipopolysaccharide; PMA=佛波酯Phorbol Myristate Acetate; 2d=2 day culture; 5hr=5 hour culture；CpG ODN 2395=未甲基化的含有CpG 结构的寡聚脱氧核苷酸。 |
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