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Proteomic Profiling of Macrophages by 2D Electrophoresis
Published on: November 4, 2014
Proteome analysis of Bordetella pertussis isolated from human macrophages
Yanina Lamberti1, Juan Hilario Cafiero1, Kristin Surmann2
1CINDEFI (UNLP CONICET La Plata), Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina.
Abstract:
Previous studies have shown that B. pertussis survives inside human macrophages in non-acidic compartments with characteristics of early endosomes. In order to gain new insight into the biology of B. pertussis survival in host cells, we have analyzed the adaptation of the bacterial proteome during intracellular infection. The proteome of B. pertussis 3 h and 48 h after infection of human macrophage-like THP-1 cells was examined by nano-liquid chromatography combined with tandem MS and compared to the protein profile of extracellular B. pertussis growing in the same cell culture medium. Compared with extracellular bacteria, almost 300 proteins out of 762 identified proteins displayed altered levels in intracellular B. pertussis. Functional analyses of the proteins displaying altered abundance revealed enrichment of proteins involved in stress response, iron uptake, cellular metabolism, transcriptional regulation, and virulence. To our knowledge, this is the first analysis of the B. pertussis proteome during adaptation to the intramacrophage environment and the data provide new clues for understanding B. pertussis adaptation and pathogenesis.
Biological Significance:
B. pertussis is a respiratory pathogen that has adapted exclusively to the human host. Despite high vaccination rates, whooping cough remains a serious threat to human health and its incidence has been increasing in recent years in vaccinated populations. The mechanisms that allow this pathogen to evade immune clearance, persist in the host, and cause a prolonged paroxysmal cough are still poorly understood. Recent studies regarding B. pertussis survival inside host cells and the cellular response to this bacterial infection indicate that B. pertussis may have an intracellular phase during infection which probably contributes to persistence and vaccine failure. In this study we provide the first global proteome profile of B. pertussis within macrophages. The data provide novel insights into the adaptive responses elicited by these bacteria for physiological adaptation to the host environment.
Insights
Bordetella pertussis (B. pertussis) adapts within human macrophages, altering nearly 300 proteins for survival. This study reveals key bacterial responses to the intracellular environment, offering insights into whooping cough pathogenesis.
Area of Science:
- Microbiology
- Immunology
- Proteomics
Background:
- Bordetella pertussis (B. pertussis) is a human respiratory pathogen responsible for whooping cough.
- Despite vaccination, B. pertussis incidence is rising, particularly in vaccinated populations.
- B. pertussis survival within host macrophages is implicated in persistence and vaccine failure.
Purpose of the Study:
- To investigate the proteomic adaptation of B. pertussis during intracellular infection of human macrophages.
- To identify bacterial proteins and pathways involved in B. pertussis survival within the host cell environment.
Main Methods:
- Proteome analysis of intracellular B. pertussis at 3 and 48 hours post-infection.
- Comparison of intracellular bacterial proteomes with extracellular B. pertussis.
- Utilized nano-liquid chromatography coupled with tandem mass spectrometry (MS/MS).
Main Results:
- Out of 762 identified proteins, nearly 300 showed altered abundance in intracellular B. pertussis compared to extracellular bacteria.
- Enrichment of proteins involved in stress response, iron uptake, metabolism, transcriptional regulation, and virulence was observed.
- This represents the first global proteome analysis of B. pertussis adapting to the intramacrophage environment.
Conclusions:
- B. pertussis undergoes significant proteomic changes to adapt to the intracellular macrophage environment.
- These adaptive responses are crucial for bacterial survival, persistence, and pathogenesis.
- Understanding these adaptations provides new insights into B. pertussis biology and potential therapeutic targets.
