TLR-6/-9 induced IL-12 secretion mediated the auto-stimulated enhanced expression of TLR-9. (A,B) Protein expression and densitometry analysis of TLR-6/-9 proteins and their downstream signaling factors, namely IRAK4, IRF7, IRF5, and IRF3, following IL-12 blocking by specific monoclonal antibodies with co-treatment of MamA2.1+ ODN2006. IL-12 cytokine release (C) and expression maturation marker CD83 (D) in THP-1 cells following IL-12 blocking under these treatment conditions. The ability of IL-12 blocked and ODN2006 stimulated THP-1 cells to induce cytotoxic CD8+T cell responses was studied by tetramer staining (E) and cytotoxicity on AU565 breast cancer cells (F). All experiments were performed in four independent replicates, and data are expressed as mean ± SD, * p < 0.05 compared with the MamA2.1 peptide treatment alone.