Figure 5

TLR2 ligands affected the fate of innate immune cells in IPEC-J2/PBMC co-culture system. IPEC-J2 cells and porcine PBMCs were co-cultured using trans-well plate. The cells were treated with B. subtilis-derived LTA (LTA-BS, 10 μg/mL) for 24 h, followed by DON treatment (2 μg/mL) for 48 h. A Attached cells in the bottom well were trypsinized and combined with the rest of the cells. Then, the cells were stained with anti-Annexin V and PI, and analyzed for the cell death using flow cytometry. The data represent means of the percentage of cells out of total cells ± SD (n = 4). B To investigate proliferation, porcine PBMCs were stained with CFDA-SE before co-culture. Then, the cells were treated with DON for 48 h with or without pretreatment with LTA-BS, and the cell proliferation was measured by flow cytometry. The degree of proliferation was shown as percentage (mean ± SD) of CFSE on CD172a⁺ monocytes. The representative result from four similar results is shown.