Ccl22

6 x 105 BHT, BHT GFP and BHT D6 cells were resuspended in 600 µl of Binding Buffer (serum-free Opti-MEM I, 4mM HEPES pH 7.4, 1% BSA) and incubated in the absence or in the presence of 1.2 nM CCL-2 or CCL-22 (R&D Systems, Minneapolis MN, USA) for 3 h at 37 °C. Additionally, 600 µl of Binding Buffer without cells were incubated in the absence or in the presence of 1.2 nM CCL-2 or CCL-2 for 3 h at 37 °C. After cell centrifugation, supernatants were collected. 1 x 106 THP-1 cells in 100 µl of serum-free Opti-MEM I (Invitrogen-Thermo Fisher Scientific, Waltham MA, USA) were placed on the polycarbonate membranes (5-mm pore size) on the bottoms of the upper compartment of the transwells (6.5 mm, Corning Life Sciences, Tewksbury MA, USA), while 600 µl of collected supernatants were added to the lower compartment. The plates were incubated at 37 °C in a 5% CO2 atmosphere saturated with H2O for 1 h. At the end of incubation, the cells at the upper side of the polycarbonate filter were mechanically removed. Cells that had migrated to the lower compartment through the filter were counted with a Neubauer chamber. The error bars represent technical replicates within a single experiment. Each experiment has been repeated at least three times.

IL-1β, IL-6, IL-23 (eBioscience), TSLP, and CCL22 (R&D) were measured in 24 h mDC, CD1c⁺ DC or BMDC culture supernatants by ELISA according to manufacturer's protocols.

Recombinant IL-21RFc or control human IgG1 (75 μg) (R&D Systems) was administered i.p. to eMCMV-infected mice 2 d before and on the day of challenge with M. tuberculosis. Mice were sacrificed 7 d later and M. tuberculosis CFU were enumerated. Recombinant mouse IL-21 (100 ng in PBS) (R&D Systems) or CCL22 (R&D Systems) as control was administered i.n. on the day of M. tuberculosis challenge, mice were sacrificed 7 d later, and M. tuberculosis CFU were enumerated.