Host-pathogen dynamics through secretome analysis of stimulated macrophages

The Toll-like receptors (TLRs) recognize different pathogen associated molecular patterns (PAMPs) and promote MyD88 dependent and independent responses. Previously, we have reported the discovery of the temporal changes in signaling cascades of macrophage proteome and secretome post-stimulation with different PAMPs. Here we present strategies to profile the secretome of TLR2-, TLR4, and TLR7- stimulated macrophages using whole pathogens were developed. Stable isotope labeling with amino acids in cell culture of macrophages was integrated with whole pathogen macrophage stimulation and subsequent targeted proteomics to quantify cytokines, chemokines, and transcription factors.