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Quantification of Cytosolic vs. Vacuolar Salmonella in Primary Macrophages by Differential Permeabilization
Published on: July 28, 2015
Differential responses of macrophages to Salmonella enterica serovars Enteritidis and Typhimurium
Masashi Okamura1, Hyun S Lillehoj, Richard B Raybourne
1Animal Parasitic Diseases Laboratory, Animal and Natural Resources Institute, Agricultural Research Service, US Department of Agriculture, Bldg 1040 BARC-East Beltsville, MD 20705, USA.
Abstract:
Macrophages are major effectors against Salmonella infection, and also transport bacteria between host tissues and provide a protected site for intracellular bacterial replication. We hypothesized that differences in chicken macrophage responses to Salmonella enterica serovar Enteritidis (SE) and serovar Typhimurium (ST) played a role in preferential infection of eggs by SE compared with ST. To test this hypothesis, we determined bacterial phagocytosis and intracellular viability and macrophage nitric oxide (NO) production following in vitro infection with SE or ST in the presence or absence of interferon-gamma (IFN-gamma). The effects of bacterial components, lipopolysaccharide (LPS), outer membrane proteins (OMP) and flagella, on NO production were also assessed. Our results showed: (1) in the presence or absence of IFN-gamma, the percentage macrophages phagocytizing SE and ST was similar; (2) the number of intracellular viable SE was significantly reduced compared with ST in the presence or absence of IFN-gamma; (3) increased macrophage necrosis was seen in the presence of IFN-gamma and ST; (4) Salmonella infection acted synergistically with IFN-gamma in induction of nitric oxide production; and (5) in the absence of IFN-gamma, macrophages produced significantly greater NO following treatment with SE outer membrane protein or flagella compared with ST OMP or flagella, while in the presence of IFN-gamma significantly less NO was produced following treatment with SE-LPS compared with ST-LPS. These results suggest that differential responses of chicken macrophages to SE versus ST may result in increased macrophage death with ST, which could result in an increased inflammatory response as compared to SE.
Insights
Chicken macrophages show different responses to Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST). ST infection leads to increased macrophage death and inflammation compared to SE, influencing Salmonella
Area of Science:
- Immunology
- Microbiology
- Veterinary Science
Background:
- Macrophages are key immune cells involved in Salmonella infection, controlling bacterial replication and spreading.
- Differential interactions between host macrophages and Salmonella serovars may influence disease pathogenesis and host specificity.
Purpose of the Study:
- To investigate the distinct responses of chicken macrophages to Salmonella enterica serovar Enteritidis (SE) and serovar Typhimurium (ST).
- To determine the role of macrophage phagocytosis, intracellular bacterial viability, and nitric oxide (NO) production in differential responses to SE and ST.
Main Methods:
- In vitro infection of chicken macrophages with SE and ST, with and without interferon-gamma (IFN-gamma).
- Assessment of bacterial phagocytosis, intracellular viability, and nitric oxide production.
- Evaluation of the effects of bacterial components (LPS, OMP, flagella) on macrophage NO production.
Main Results:
- Macrophage phagocytosis rates for SE and ST were similar, with or without IFN-gamma.
- Intracellular viable SE were significantly reduced compared to ST, irrespective of IFN-gamma presence.
- Salmonella infection synergized with IFN-gamma to induce nitric oxide production.
- ST infection, particularly with IFN-gamma, induced increased macrophage necrosis and NO production compared to SE.
Conclusions:
- Chicken macrophage responses differ significantly between SE and ST infections.
- ST infection may lead to increased macrophage death and a heightened inflammatory response compared to SE.
- These differential immune responses could contribute to the distinct disease tropisms observed between SE and ST.

