Next, we measured changes in immune cells, such as macrophages and Th17 cells, and immunosuppressive cells, including CD4+CD25+ regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs), using flow cytometry and immunohistochemistry to determine potential relationships between both forms of TNF-α and these cells in the mouse AOM/DSS-induced CAC model. Here, CD4 is linked to infectious otitis media.