Interleukin 4 (il 4)

Bone marrow cells were isolated from the femur and tibia of C57BL/6 mice. Collected cells were treated with ACK buffer (Cambrex Bio Sciences, East Rutherford, NJ, usA) to lyse the red blood cells. Resulting cells were differentiated for 7 d at 1 × 10⁶ cells/ml in McCoy’s medium, supplemented with sodium bicarbonate (2.2 g/l), 10% FBS, antibiotics (100 µg/ml penicillin G and 50 µg/ml gentamycin), mouse recombinant GM-CSF (10 ng/ml) and IL-4 (10 ng/ml) (Cell Sciences, Canton, MA, USA).^{33 (link)} At d 7, non-adherent cells were collected and positively selected for CD11c⁺ cells using CD11c Micro Beads (Miltenyi Biotec, San Diego, CA, USA), resulting in >98% CD11c⁺ population. Bone marrow-derived DCs (BMDCs) were cultured in DMEM complete medium supplemented with 10% FBS (Sigma-Aldrich) at 37°C with 5% CO₂. Experiments to obtain mouse-derived cells were approved by the University of Texas-Houston Health Science Center animal welfare committee (HSC-AWC-10-087 and HSC-AWC-11-158).

Resulting cells were differentiated for 7 days at 1×10⁶ cells/mL in McCoy’s medium, supplemented with sodium bicarbonate (2.2 g/L), 10% FBS, antibiotics (100 μg/mL Penicillin G, and 50 μg/mL Gentamycin), mouse recombinant GM-CSF (10 ng/mL), and IL-4 (10 ng/mL) (Cell Sciences). At day 7, non-adherent cells were collected and positively selected for CD11c⁺ cells using CD11c MicroBeads (Miltenyi Biotec), resulting in >98% CD11c⁺ population. Bone marrow derived dendritic cells were cultured in DMEM complete medium supplemented with 10% FBS (Sigma-Aldrich) at 37°C with 5% CO₂.