Proteome and secretome analysis reveals differential post-transcriptional regulation of Toll-like receptor responses. Mus musculus

The innate immune system is the organism’s first line of defense against pathogens. Pattern recognition receptors (PRRs) are responsible for sensing the presence of pathogen-associated molecules. The prototypic PRRs, the membrane-bound receptors of the Toll-like receptor (TLR) family, recognize pathogen-associated molecular patterns (PAMPs) and initiate an innate immune response through signaling pathways that depend on the adaptor molecules MyD88 and TRIF. Deciphering the differences in the complex signaling events that lead to pathogen recognition and initiation of the correct response remains challenging. Here we report the discovery of temporal changes in the protein signaling components involved in innate immunity. Using an integrated strategy combining unbiased proteomics, transcriptomics and macrophage stimulations with three different PAMPs, we identified differences in signaling between individual TLRs and revealed specifics of pathway regulation at the protein level. Overall design: RAW264.7 cells were treated with individual TLR ligands (100 ng/mL LPS, 1 μM P3C, and 1 μM R848 for stimulation of TLR4, TLR2, and TLR7, respectively) for either 1, 2 or 4 h. Two replicates for each condition.