TABLE 1.
Differential expression of genes for selected cytokines and chemokines after stimulation with the recombinant A. fumigatus antigens Aspf1 and Crf1a
| Gene | Differential expression (fold) at the indicated times after stimulation with:
|
|||||
|---|---|---|---|---|---|---|
| Aspf1
|
Crf1
|
|||||
| 6 h | 24 h | 48 h | 6 h | 24 h | 48 h | |
| TNF-α | 8.2 | 12 | 22 | — | — | 3 |
| IL-8 | 30 | 72 | 275 | 2.1 | 2.8 | 2.9 |
| IL-23p19 | 7.5 | 2.2 | — | ND | ND | ND |
| CXCL10 | 39 | 60 | 27 | 2.3 | 2.9 | 3.1 |
| CCL20 | 247 | 299 | 314 | — | 3.1 | — |
| IL-12p35 | ND | 14 | ND | ND | ND | ND |
| IL-10 | 0.6 | 0.8 | — | ND | ND | ND |
| IL-1β | ND | 7.4 | ND | ND | ND | ND |
DCs were stimulated for 6, 24, or 48 h with A. fumigatus antigens; and the levels of expression of the genes for TNF-α, IL-8, IL-23, CXCL10, and CCL20 were determined by quantitative reverse transcription-PCR. The level of gene expression was normalized against the level of expression of h-ALAS by evaluating the crossing point values for each sample by using the equation 2(ΔCP for target gene − ΔCP for h-ALAS), where ΔCP is the change in the crossing point (CP) value (7, 22). For each time point, the levels of gene expression by DCs cocultivated with A. fumigatus antigens were compared to the levels of gene expression by unstimulated cells (and are given as ratios). All experiments were repeated with blood from at least three independent blood donors, and the mean level of upregulation is indicated. —, unchanged expression, defined as a <2-fold change and a >0.8-fold change; ND, not determined.