Hccl 2

The concentrations of cytokines hIL-33, hIL-1α, hCCL-2 (R&D systems), hTSLP, mIL-33, mTSLP, mIL-4 and mIL-1α (eBiosciences) in the cell supernatant, Lung homogenate and in the BALF were estimated using paired antibodies following manufacturer’s instructions.

MSCs (5000 cells/ml) were plated in the lower chamber of trans-well plates (5 μm) and cultured for 24 h. Leukemia cells, 0.4 × 10⁶ (2 × 10⁶/ml), were added in the insert and cultured for 6 to 24 h. The percentage of migration was defined as viable CD45⁺ cells in the lower chamber/total viable leukemia cells in insert and lower chamber. For migration toward ligands, chemokines were added to the lower chamber and the migration of leukemia cells was measured at 4 to 6 h. CCL2, CCL4, and CXCL12 ligands, neutralizing antibodies for CCL2 (hCCL2) and CCL4 (hCCL4) and antagonist for CCR5 (maraviroc) were purchased from R & D Systems Inc. (Minneapolis, MN). Neutralizing antibodies for CCR2 and CCR5 were purchased from Abcam (Cambridge, MA), and antagonist for CCR2 was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX).