PhagoCyt enables isolation of Salmonella-containing phagosomes and mapping of host and bacteria remodelling of the intracellular niche.

Flow cytometry-based isolation plus ultrasensitive proteomics enables high-throughput profiling of pure Salmonella-containing phagosomes from macrophages, quantifying more proteins than prior methods at 30 min and 4 hours.

• Why it matters: Pure pathogen-containing phagosomes are hard to isolate, limiting how well host and bacterial remodeling of the intracellular niche can be mapped during infection.
• What they did: Macrophages were infected with wild-type Salmonella enterica serovar Typhimurium or a ΔphoP mutant, and phagosomal and bacterial proteomes were analyzed by flow cytometry isolation and ultrasensitive proteomics at 30 min and 4 hours post-infection.
• The result: The proteomes show PhoP-dependent virulence factor induction and identify novel putative proteins shaping STM’s intracellular niche, while bacteria-containing phagosomes recruit mitochondrial membrane linked to reactive oxygen species production.

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