Table 1. Age-dependent TLRA-induced cytokine production.
Averaged baseline (vehicle control condition) and maximal cytokine levels for each study age group and agonist, as well as the agonist concentration (ng/ml for MPLA and GLA; μM for R848 and μg/ml for CpG) required for the maximal cytokine response. p-Values of 2-way ANOVA comparing the age groups are indicated.
| Agonist | Cytokine | Newborn Baseline |
Newborn Max resp. |
Newborn [Agonist] |
Infant Baseline |
Infant Max resp. |
Infant [Agonist] |
Adult Baseline |
Adult Max resp. |
Adult [Agonist] |
RM-ANOVA NB vs IN |
RM-ANOVA IN vs AD |
RM-ANOVA NB vs AD |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| MPLA | CXCL10 | 23.40 | 94.84 | 1000.00 | 30.60 | 947.78 | 1000.00 | 14.39 | 1900.63 | 1000.00 | *0.0188 | 0.4403 | *0.0152 |
| CCL4 | 30.51 | 5091.51 | 1000.00 | 43.433 | 4767.39 | 1000.00 | 83.35 | 5084.25 | 1000.00 | ***0.0006 | 0.7342 | ***0.0009 | |
| CCL5 | 2521.03 | 4649.00 | 1000.00 | 2861.61 | 4514.48 | 1000.00 | 1951.22 | 2517.60 | 1000.00 | 0.1259 | ***<0.0001 | ***<0.0001 | |
| IFN-γ | 1.83 | 8.85 | 125.00 | 13.93 | 103.76 | 3.91 | 4.73 | 78.23 | 1000.00 | *0.0130 | *0.2260 | **0.0019 | |
| TNF | 20.44 | 73.82 | 1000.00 | 23.71 | 225.35 | 1000.00 | 40.73 | 201.57 | 1000.00 | ***0.0002 | 0.6236 | **0.0053 | |
| IL-12p70 | 1.94 | 4.56 | 1000.00 | 3.82 | 21.28 | 1000.00 | 1.68 | 7.85 | 1000.00 | **0.0033 | **0.0033 | 0.9055 | |
| IFN-α | 25.92 | 40.98 | 1000.00 | 21.43 | 1315.04 | 31.25 | 13.35 | 28.82 | 500.00 | **0.0056 | **0.0047 | ***<0.0001 | |
| IL-1β | 1.46 | 127.82 | 1000.00 | 1.89 | 1189.93 | 250.00 | 1.22 | 217.29 | 1000.00 | *0.0177 | 0.0770 | 0.0279 | |
| GM-CSF | 2.74 | 10.29 | 1000.00 | 3.44 | 108.49 | 125.00 | 3.30 | 12.27 | 500.00 | 0.1149 | 0.0935 | 0.4930 | |
| IL-6 | 12.59 | 2847.25 | 1000.00 | 8.23 | 2353.76 | 1000.00 | 13.17 | 2514.88 | 1000.00 | 0.0658 | *0.0176 | 0.4060 | |
| IL-12p40 | 5.13 | 21.22 | 1000.00 | 16.51 | 36.75 | 1000.00 | 4.21 | 56.18 | 1000.00 | **0.0023 | *0.0457 | 0.1138 | |
| IL-10 | 11.54 | 96.39 | 1000.00 | 5.84 | 128.96 | 1000.00 | 9.18 | 112.81 | 1000.00 | *0.0144 | 0.8903 | *0.0148 | |
| CCL2 | 114.51 | 9550.86 | 1000.00 | 301.78 | 5878.50 | 500.00 | 219.85 | 5652.82 | 500.00 | ***0.0002 | 0.6236 | **0.0053 | |
| CXCL8 | 63.98 | 4628.43 | 1000.00 | 34.15 | 3806.51 | 500.00 | 63.73 | 1299.08 | 1000.00 | 0.1669 | ***<0.0001 | ***<0.0001 | |
| PGE2 | 174.41 | 294.51 | 1000.00 | 74.34 | 117.64 | 250.00 | 16.65 | 57.48 | 62.50 | ***<0.0001 | *0.0236 | ***<0.0001 | |
| GLA | CXCL10 | 25.02 | 245.67 | 1000.00 | 12.89 | 2481.00 | 250.00 | 14.66 | 3397.00 | 250.00 | ***<0.0001 | ***<0.0001 | 0.0703 |
| CCL4 | 36.20 | 9338.89 | 1000.00 | 52.00 | 6828.87 | 1000.00 | 40.33 | 6420.38 | 1000.00 | 0.3209 | ***<0.0001 | ***<0.0001 | |
| CCL5 | 2579.37 | 5716.69 | 1000.00 | 1513.69 | 6242.68 | 1000.00 | 918.29 | 2854.60 | 1000.00 | 0.1839 | ***<0.0001 | ***<0.0001 | |
| IFN-γ | 1.83 | 31.31 | 1000.00 | 13.93 | 46.25 | 31.25 | 4.73 | 275.62 | 1000.00 | 0.0515 | 0.5833 | ***<0.0001 | |
| TNF | 41.20 | 90.59 | 1000.00 | 101.64 | 700.63 | 1000.00 | 22.15 | 351.05 | 1000.00 | ***<0.0001 | ***0.0005 | ***<0.0001 | |
| IL-12p70 | 1.80 | 6.00 | 1000.00 | 2.08 | 30.42 | 31.25 | 1.21 | 89.54 | 250.00 | **0.0062 | *0.0284 | *0.0237 | |
| IFN-α | 21.06 | 42.69 | 1000.00 | 21.06 | 1322.14 | 250.00 | 11.84 | 34.85 | 500.00 | **0.0091 | **0.0080 | **0.0076 | |
| IL-1β | 2.41 | 1425.75 | 1000.00 | 8.91 | 1562.25 | 250.00 | 3.63 | 1667.05 | 1000.00 | 0.2228 | 0.0611 | **0.0079 | |
| GM-CSF | 1.48 | 22.10 | 1000.00 | 3.34 | 39.55 | 125.00 | 3.07 | 11.50 | 1000.00 | 0.0784 | *0.0134 | 0.1961 | |
| IL-6 | 18.13 | 9245.33 | 1000.00 | 9.31 | 4924.44 | 1000.00 | 13.75 | 6221.11 | 1000.00 | ***0.0001 | **0.0022 | ***<0.0001 | |
| IL-12p40 | 5.63 | 63.49 | 1000.00 | 6.20 | 70.13 | 1000.00 | 4.75 | 160.11 | 1000.00 | *0.0153 | 0.2148 | ***0.0005 | |
| IL-10 | 9.68 | 705.00 | 1000.00 | 8.52 | 475.31 | 1000.00 | 18.84 | 438.13 | 1000.00 | 0.2141 | **0.0017 | **0.0026 | |
| CCL2 | 177.07 | 9909.23 | 1000.00 | 575.10 | 5726.38 | 125.00 | 147.12 | 4275.25 | 125.00 | ***<0.0001 | ***0.0005 | ***<0.0001 | |
| CXCL8 | 66.86 | 8013.28 | 1000.00 | 19.59 | 3634.63 | 125.00 | 30.16 | 1604.63 | 1000.00 | *0.0316 | ***<0.0001 | ***<0.0001 | |
| PGE2 | 174.41 | 1817.03 | 1000.00 | 74.34 | 310.54 | 1000.00 | 20.29 | 322.76 | 1000.00 | ***<0.0001 | 0.1184 | ***<0.0001 | |
| R848 | CXCL10 | 27.01 | 6207.86 | 0.20 | 13.12 | 6982.85 | 3.13 | 16.80 | 4629.75 | 1000.00 | 0.0079 | 0.1352 | 0.2075 |
| CCL4 | 64.83 | 11133.67 | 100.00 | 1303.32 | 7509.34 | 6.25 | 57.45 | 4931.45 | 1000.00 | **0.0018 | 0.0838 | ***<0.0001 | |
| CCL5 | 2503.74 | 6087.67 | 100.00 | 2812.48 | 5796.27 | 3.13 | 808.00 | 3403.80 | 12.50 | 0.6063 | ***<0.0001 | ***<0.0001 | |
| IFN-γ | 1.83 | 649.88 | 12.50 | 13.93 | 1205.75 | 25.00 | 4.73 | 2304.30 | 25.00 | *0.0448 | ***<0.0001 | ***<0.0001 | |
| TNF | 22.12 | 3880.15 | 25.00 | 106.54 | 2027.31 | 12.50 | 22.13 | 1967.20 | 25.00 | ***0.0002 | 0.4638 | ***<0.0001 | |
| IL-12p70 | 2.00 | 10.68 | 50.00 | 2.43 | 30.77 | 100.00 | 1.67 | 128.82 | 12.50 | ***<0.0001 | ***<0.0001 | ***<0.0001 | |
| IFN-α | 22.99 | 116.98 | 1.56 | 28.69 | 589.38 | 1.56 | 13.86 | 167.18 | 6.25 | ***<0.0001 | ***<0.0001 | **0.0078 | |
| IL-1β | 5.12 | 8790.84 | 50.00 | 20.44 | 3803.85 | 12.50 | 5.29 | 5472.60 | 12.50 | ***<0.0001 | 0.123 | ***<0.0001 | |
| GM-CSF | 2.07 | 29.91 | 50.00 | 4.20 | 34.58 | 12.50 | 4.42 | 32.05 | 50.00 | 0.5519 | 0.7408 | 0.4248 | |
| IL-6 | 30.93 | 10156.18 | 50.00 | 193.22 | 6085.32 | 6.25 | 14.45 | 8518.30 | 12.50 | ***<0.0001 | **0.0033 | ***<0.0001 | |
| IL-12p40 | 7.70 | 407.00 | 12.50 | 9.47 | 478.94 | 12.50 | 4.25 | 1150.25 | 6.25 | 0.8557 | ***<0.0001 | ***<0.0001 | |
| IL-10 | 10.67 | 2195.38 | 12.50 | 70.89 | 1652.82 | 6.25 | 7.67 | 1381.13 | 6.25 | ***0.0005 | 0.3275 | ***<0.0001 | |
| CCL2 | 268.44 | 10275.11 | 50.00 | 1313.88 | 6216.66 | 1.56 | 248.37 | 5212.38 | 1.56 | ***0.0002 | 0.4638 | ***<0.0001 | |
| CXCL8 | 96.57 | 9207.12 | 50.00 | 70.79 | 1495.50 | 12.50 | 36.51 | 467.75 | 12.50 | ***<0.0001 | ***<0.0001 | ***<0.0001 | |
| PGE2 | 174.41 | 5132.93 | 12.50 | 74.34 | 116.93 | 100.00 | 25.26 | 74.42 | 6.25 | ***0.0002 | 0.8489 | ***0.0002 | |
| ODN | CXCL10 | 24.19 | 1698.78 | 0.10 | 10.60 | 3783.92 | 6.25 | 46.41 | 1116.45 | 12.50 | 0.2249 | ***0.0001 | *0.0148 |
| CCL4 | 39.13 | 1287.25 | 50.00 | 28.26 | 1229.00 | 12.50 | 346.63 | 1401.43 | 12.50 | 0.0450 | 0.2523 | 0.5270 | |
| CCL5 | 3437.82 | 6827.82 | 12.50 | 1567.06 | 6071.13 | 25.00 | 993.43 | 2618.82 | 6.25 | 0.7905 | ***0.0001 | ***0.0001 | |
| IFN-γ | 1.83 | 92.21 | 50.00 | 13.93 | 65.93 | 0.10 | 4.73 | 26.52 | 25.00 | 0.7653 | **0.0057 | *0.0277 | |
| TNF | 39.93 | 44.79 | 0.10 | 61.21 | 159.79 | 0.39 | 27.06 | 77.77 | 1.56 | **0.0015 | *0.0231 | **0.0075 | |
| IL-12p70 | 2.13 | 3.63 | 50.00 | 1.74 | 8.33 | 12.50 | 1.67 | 2.34 | 12.50 | *0.0120 | ***0.0001 | ***0.0007 | |
| IFN-α | 30.54 | 31.15 | 3.13 | 30.55 | 160.14 | 6.25 | 9.85 | 33.29 | 12.50 | ***0.0007 | ***<0.0001 | ***0.0004 | |
| IL-1β | 1.86 | 1728.29 | 25.00 | 8.71 | 115.53 | 0.20 | 5.24 | 17.80 | 50.00 | *0.0331 | **0.0058 | *0.0156 | |
| GM-CSF | 2.91 | 7.32 | 25.00 | 1.83 | 7.28 | 12.50 | 3.17 | 6.11 | 12.50 | 0.4665 | 0.1486 | 0.3776 | |
| IL-6 | 12.14 | 1692.84 | 25.00 | 75.18 | 787.49 | 0.20 | 28.60 | 41.03 | 50.00 | *0.0408 | ***0.0010 | **0.0018 | |
| IL-12p40 | 6.87 | 98.65 | 6.25 | 4.90 | 31.82 | 50.00 | 26.01 | 26.01 | 0.00 | *0.0115 | **0.0021 | *0.0156 | |
| IL-10 | 9.78 | 287.52 | 6.25 | 27.89 | 63.21 | 12.50 | 7.37 | 29.69 | 12.50 | **0.0029 | ***0.0009 | ***0.0001 | |
| CCL2 | 121.53 | 8958.76 | 50.00 | 850.22 | 3622.37 | 12.50 | 231.66 | 944.98 | 25.00 | ***0.0001 | ***0.0001 | ***0.0001 | |
| CXCL8 | 79.18 | 5059.90 | 50.00 | 54.04 | 659.78 | 50.00 | 27.36 | 154.74 | 50.00 | ***<0.0001 | ***<0.0001 | *0.0282 | |
| PGE2 | 174.41 | 286.31 | 50.00 | 74.34 | 157.53 | 50.00 | 25.03 | 41.16 | 6.25 | ***<0.0001 | 0.0597 | ***<0.0001 |
*
p<0.05,
**
p<0.01,
***
p<0.001