Anti human il 10

To perform flow cytometry and Western blot, the following materials were used: goat anti-mouse IgG antibody, peroxidase conjugated, H+L (Millipore, AP124P), anti-human IL-10 (Peprotech, 500-M86), recombinant hIL-10 (Peprotech, 200-10), and western blotting luminol reagent (Santa Cruz, sc-2048).

mDCs were stimulated with 0.1, 1, or 10 μg/ml of irradiated M. leprae, and its membrane (MLMA) and cytosol fractions (MLSA) (BEI Resources [NIH/ATCC], Manassas, VA, USA, 200 μM of the IDO inhibitor 1‐methyl‐D‐trytophan (1‐MT, SigmaAldrich, Saint Louis, MO, USA), 500 μg/ml of the anti‐human TNF (PeproTech), 100 μg/ml of the anti‐human IL‐10 (PeproTech), 10 μg/ml of the TLR2 agonist Pam3Cys‐Ser‐(Lys)4 (SigmaAldrich), 100 μM of the inhibitory peptide for MyD88 homodimerization (IMGENEX, San Diego, CA, USA), or 1 μg/ml of Mab‐mTLR2 (InvivoGen, San Diego, CA, USA), and incubated for 24 h at 37°C in 5% CO₂. M. leprae and its fractions were tested for purity and the absence of endotoxin. According to the limulus amebocyte lysate assay (Lonza, Basel, Switzerland), all stimuli used for in vitro cultures were shown to contain less than 0.1 U/ml endotoxin.