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Flow cytometry-based isolation of Salmonella-containing phagosomes combined with ultra-sensitive proteomics reveals novel insights into host-pathogen interactions
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Flow cytometry-based isolation of Salmonella-containing phagosomes combined with ultra-sensitive proteomics reveals novel insights into host-pathogen interactions
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Paper
Flow cytometry-based isolation of Salmonella-containing phagosomes combined with ultra-sensitive proteomics reveals novel insights into host-pathogen interactions
2025
Overview
Macrophages engulf pathogens into dynamic phagosomes, which many bacteria manipulate for survival. However, isolating pure pathogen-containing phagosomes remains challenging. Here, we developed a novel flow cytometry-based isolation and ultrasensitive proteomics approach to analyse phagosomal and bacterial proteomes from macrophages infected with wild-type (WT) Salmonella enterica serovar Typhimurium (STM) or a ΔphoP mutant at 30 min and 4 hrs post-infection. Our approach provides higher throughput, requires lower cell numbers and quantifies more proteins than previous techniques. Our data reveals key host-pathogen interactions, showing induction of PhoP-dependent virulence factors and novel putative proteins that shape STM’s intracellular niche. Moreover, our data indicates that bacteria-containing phagosomes recruit mitochondrial membrane for production of reactive oxygen species. These findings provide new insights into Salmonella’s manipulation of phagosomal maturation and intracellular niche formation.
Publisher
Cold Spring Harbor Laboratory
Subject
