Essential to bacterial pathogenesis, Salmonella enterica serovar Typhimurium (S. Typhimurium) has evolved the capacity to quickly sense and adapt to specific intracellular environment within distinct host cells. Here we examined S. Typhimurium proteomic remodeling within macrophages, allowing direct comparison with our previous studies in epithelial cells. In addition to many shared features, our data revealed proteomic signatures highly specific to one type of host cells. Notably, intracellular S. Typhimurium differentially regulates the two type III secretion systems (T3SSs) far quicker in macrophages than in epithelial cells, so do bacterial flagellar and chemotaxis systems degenerate. Importantly, our comparative analysis uncovered vast induction of bacterial histidine biosynthesis in macrophages but not in epithelial cells. Targeted metabolomic measurements revealed markedly lower histidine levels within macrophages. Intriguingly, further functional studies established that defective histidine biosynthesis (due to a hisG mutation) renders the bacterium (strain SL1344) hypersensitive to intracellular shortage of this amino acid. Indeed another S. Typhimurium strain 14028s with a fully functional biosynthetic pathway exhibited only minor induction of the his operon within infected macrophages. Our work thus reveals novel insight into S. Typhimurium adaptation mechanisms within distinct host cells and also provides an elegant paradigm where proteomic profiling of intracellular pathogens is utilized to discriminate specific host environment (e.g., on nutrient availability).