Table 1.
Cocktail compositions and the cocktail stimulated DC expression pattern for selected cytokines and chemokines on cytokine arrays
| Cocktail | Composition | Secreted cytokines and chemokines | ||
|---|---|---|---|---|
| Weak induction (1-10 fold) |
Modest induction (10-100 fold) |
Strong induction (> 100 fold) |
||
| LPS | LPS (0,1ug/mL) | IL-8, CCL2, CCL4, CXCL5, CXCL10, GM-CSF | IL-10, CCL20, CXCL1-3 | IL-6, IL-12, TNFα, VEGF, CCL8, CCL15, CXCL1 |
| Cocktail 1 | IFNγ (20ng/mL)+ TNFα (50ng/mL)+ Poly I:C 12,5 ug/mL + IL-1β (10ng/mL) + IFNα (6ng/mL) | IL-8, CCL2, CCL4, CCL5, CXCL5, CXCL10 | IL-10, CCL20, CXCL1-3, GM-CSF | IL-6, IL-12, TNFα, VEGF, CCL8, CCL15, CXCL1 |
| Cocktail 2 | IFNγ (20ng/mL)+ Poly I:C 12,5 ug/mL + IL-1β (10 ng/mL) + IFNα (6ng/mL) | IL-8, CCL2, CCL4, CCL5, CXCL5, CXCL10 |
IL-10, CCL20, CXCL1-3, GM-CSF | IL-6, IL-12, TNFα, VEGF, CCL8, CCL15, CXCL1 |
| Cocktail 3 | IFNγ (20ng/mL)+TNFα (50ng/mL)+ peptidoglycan (10ug/mL) | IL-8, CCL2, CCL4, CCL5, CXCL5 | IL-10, CCL20, CXCL1-3, CXCL10, GM-CSF | IL-6, IL-12, TNFα, VEGF, CCL8, CCL15, CXCL1 |
| Cocktail 4 | LPS (0,1ug/mL)+ IFNγ (20ng/mL) | IL-8, CCL2, CCL4, CCL5, CXCL5, GM-CSF | IL-10, CCL20, CXCL1-3, CXCL10 | IL-6, IL-12, TNFα, VEGF, CCL8, CCL15, CXCL1 |
| Cocktail 5 | LPS (0,1ug/mL)+ IFNγ (20ng/mL)+ TNFα (50ng/mL) | IL-8, CCL2, CCL4, CCL5, CXCL5, GM-CSF | IL-10, CCL20, CXCL1-3, CXCL10 | IL-6, IL-12, TNFα, VEGF, CCL8, CCL15, CXCL1 |
| Cocktail 6 | R848 (1,0ug/mL) | IL-8, CCL2, CCL4, CCL5, CXCL10 | IL-10, CCL15, CCL20, CXCL1-3, CXCL5, GM-CSF | IL-6, IL-12, TNFα, VEGF, CCL8, CXCL1 |
| Cocktail 7 | R848 (1,0ug/mL) + IFNγ (20ng/mL) | IL-8, CCL2, CCL4, CCL5, CCL20, CXCL5, GM-CSF | IL-10, CCL15, CCL20, CXCL1-3, CXCL10 | IL-6, IL-12, TNFα, VEGF, CCL8, CXCL1 |
| Cocktail 8 | R848 (1,0ug/mL) + poly I:C (10ug/mL) | IL-8, CCL2, CCL4, CCL5, CXCL5, CXCL10 | IL-10, CCL20, CXCL1-3, GM-CSF | IL-6, IL-12, TNFα, VEGF, CCL8, CCL15, CXCL1 |
| Cocktail 9 | IFNγ (20ng/mL) + IL-1β (10ng/mL) | IL-8, CCL2, CCL4, CCL5, CXCL5, GM-CSF | IL-10, CCL15, CCL20, CXCL1-3, CXCL10 | IL-6, IL-12, TNFα, VEGF, CCL8, CXCL1 |
Table 1 shows the composition of cocktails combined by TLR-agonists and cytokines. TLR agonists were peptidoglycan (TLR2), Poly I:C (TLR3), LPS (TLR4/CD14/MD2), R848 (TLR7/8) and cytokines IFNγ, TNFα, IL-1β, IFNα. Secreted cytokines and chemokines were determined by combination of conditioned media for treatment of immature DCs from four different donors in order to obtain information for a general response and reduce single donor-variations. Signal intensities were semi-quantitatively calculated using the ImageJ software, and the fold of induction compared to untreated cells. Induced cytokines and chemokines were classified into weakly induced proteins (1-10 fold induced vs untreated), modestly induced proteins (10-100 fold induced vs untreated controls), and strongly induced proteins (more than 100 fold vs untreated controls).