Pooled cervical, axial, and inguinal lymph node cells (106/well) from naïve A/J, BALB/c, and C57BL/6 mice were extracted. Cells obtained were exposed to IL-2 (100 UI/mL, Peprotech) alone or with the addition of plate-immobilized anti-CD3e (2 μg/mL, BD), anti-CD28 (2 μg/mL, BD), and TGF-β (2 ng/mL, Peprotech) for 72 h at 37°C and 5% CO2 [25 (link)]. Cells were retrieved for immunophenotyping, and when relevant, supernatant was evaluated for detection of cytokine secretion, as described on the previous section.
mLN cells from OVA-challenged mice were exposed in vitro to IL-2 (100 UI/mL) alone or with the addition of OVA (0.5 mg/mL) with or without TGF-β (2 ng/mL) for 72 h at 37°C and 5% CO2.
mLN cells from OVA-challenged mice were exposed in vitro to IL-2 (100 UI/mL) alone or with the addition of OVA (0.5 mg/mL) with or without TGF-β (2 ng/mL) for 72 h at 37°C and 5% CO2.
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