Toll-like receptors (TLRs) are key mediators of both innate and adaptive immunity by recognizing and eliciting responses to invading pathogens. The activation of TLRs must be stringently controlled in order to avoid exaggerated expression of signaling components as well as pro-inflammatory cytokines that can devastate the host, resulting in chronic inflammatory diseases, autoimmune disorders and aid in the pathogenesis of TLR-associated diseases. Therefore, it is essential that negative regulators act at multiple levels within TLR signaling cascades in order to synchronize the activation and negative regulation of signal transduction to limit potentially harmful immunological consequences. A summary of the various mechanisms employed by negative regulators of TLRs signaling to ensure the appropriate modulation of both immune and inflammatory responses was provided.