Single-Cell and Time-Resolved Profiling of Intracellular Salmonella Metabolism in Primary Human Cells.
Jiabao XuLorena Preciado-LlanesAnna AulicinoChristoph Martin DeckerMaren DepkeManuela Gesell SalazarFrank SchmidtAlison SimmonsWei E HuangPublished in: Analytical chemistry (2019)
The intracellular pathogen Salmonella enterica has evolved an array of traits for propagation and invasion of the intestinal layers. It remains largely elusive how Salmonella adjusts its metabolic states to survive inside immune host cells. In this study, single-cell Raman biotechnology combined with deuterium isotope probing (Raman-DIP) have been applied to reveal metabolic changes of the typhoidal Salmonella Typhi Ty2, the nontyphoidal Salmonella Typhimurium LT2, and a clinical isolate Typhimurium D23580. By initially labeling the Salmonella strains with deuterium, we employed reverse labeling to track their metabolic changes in the time-course infection of THP-1 cell line, human monocyte-derived dendritic cells (MoDCs) and macrophages (Mf). We found that, in comparison with a noninvasive serovar, the invasive Salmonella strains Ty2 and D23580 have downregulated metabolic activity inside human macrophages and dendritic cells and used lipids as alternative carbon source, perhaps a strategy to escape from the host immune response. Proteomic analysis using high sensitivity mass spectrometry validated the findings of Raman-DIP analysis.
Keyphrases
- listeria monocytogenes
- dendritic cells
- escherichia coli
- single cell
- immune response
- endothelial cells
- mass spectrometry
- rna seq
- high throughput
- genome wide
- regulatory t cells
- high resolution
- pluripotent stem cells
- raman spectroscopy
- candida albicans
- induced pluripotent stem cells
- toll like receptor
- high performance liquid chromatography
- tandem mass spectrometry
- inflammatory response
- liquid chromatography
- high density