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Culture conditions
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Intracellular expression and production of the cytokines IFN-γ and IL-4
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 |  |
Control group (n = 10)
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NP women (n = 8)
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HA women (n = 8)
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 |  |
Value
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SI
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Value
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SI
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Value
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SI
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CD3+ IFN-γ+ (%) | 0 | 1.5 ± 0.2 |  | 1.8 ± 0.5 |  | 1.8 ± 0.5 |  |
 | + DCs | 5.9 ± 0.6 | 5.2 ± 0.9 | 1.5 ± 0.27** | 1.1 ± 0.2** | 2.5 ± 0.3**#
| 2.0 ± 0.4**#
|
CD3+ IL-4+ (%) | 0 | 1.7 ± 0.1 |  | 2.6 ± 0.4 |  | 3.3 ± 0.5 |  |
 | + DCs | 2.1 ± 0.4 | 1.3 ± 0.2 | 8.6 ± 1.0** | 3.5 ± 0.4** | 7.4 ± 1.0** | 2.4 ± 0.5* |
IFN-γ (pg/ml) | 0 | 18.2 ± 5.7 |  | 16.5 ± 2.8 |  | 16 ± 3.0 |  |
 | + DCs | 353 ± 41 | 22.2 ± 3.1 | 63.4 ± 27.8** | 3.4 ± 0.9** | 218 ± 69* #
| 13.5 ± 3.1*#
|
IL-4 (pg/ml) | 0 | 1.0 ± 0.08 |  | 1.0 ± 0.09 |  | 1.0 ± 0.12 |  |
 | + DCs | 2.0 ± 1.0 | 2.0 ± 1.0 | 4.6 ± 2.2 | 4.6 ± 2.2 | 5.4 ± 2.0* | 5.4 ± 2.0* |
- Donor MNCs were co-cultured with allogenic IFN-α-DCs from non-pregnant women (control), normal pregnant women (NP), or hyperandrogenia pregnant women (HA) for 72 h at an MNC-to-DC ratio of 10:1. For the final 18 h of culture, 10 μg/ml Brefeldin A was added. Intracellular expression of IFN-γ and IL-4 was detected within the CD3+ gate by flow cytometry. To determine the levels of cytokine production by MNCs, cell-free supernatants were collected after 5 days of co-culturing donor MNCs and allogenic IFN-α-DCs from the control group, NP-women, or HA-women (at a MNC-to-DC ratio of 10:1) and were assessed for production of IL-4 and IFN-γ using specific ELISA kits. Data are presented as mean (± SE) cytokine concentrations (pg/ml) or the percentage of CD3+ cells from the indicated number of independent experiments. SI indicates the stimulation index, which was calculated as the ratio of cytokine concentrations (pg/ml) or intracellular cytokine expression (%) by MNCs in the presence or absence of DCs. *, pU < 0.05, **, pU < 0.01, compared to the control women group; #, pU < 0.05, compared to the NP-women group. Significant differences were detected using the Mann–Whitney U test.