A flow cytometric assay to quantify in vivo bacterial uptake by alveolar macrophages

Pitz, Adam; Perry, Greg A.; Jensen-Smith, Heather; Gentry-Nielsen, Martha J.

doi:10.1016/j.mimet.2010.02.005

Cited by 7 publications

(6 citation statements)

References 11 publications

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“…Our in vitro phagocytosis results by using murine macrophages warrant further investigation through the evaluation of phagocytosis of bacteria by monocytes/macrophages in mice during infection (35) following pretreatment of animals with AHLs.…”

Section: Vol 79 2011 Homoserine Lactones Enhance Innate Immunity Inmentioning

confidence: 92%

Immunomodulatory and Protective Roles of Quorum-Sensing Signaling Molecules N -Acyl Homoserine Lactones during Infection of Mice with Aeromonas hydrophila

et al. 2011

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Aeromonas hydrophila leads to both intestinal and extraintestinal infections in animals and humans, and the underlying mechanisms leading to mortality are largely unknown. By using a septicemic mouse model of infection, we showed that animals challenged with A. hydrophila die because of kidney and liver damage, hypoglycemia, and thrombocytopenia. Pretreatment of animals with quorum-sensing-associated signaling molecules N-acyl homoserine lactones (AHLs), such as butanoyl and hexanoyl homoserine lactones (C 4 -and C 6 -HSLs), as well as N-3-oxododecanoyl (3-oxo-C 12 )-HSL, prevented clinical sequelae, resulting in increased survivability of mice. Since little is known as to how different AHLs modulate the immune response during infection, we treated mice with the above AHLs prior to lethal A. hydrophila infection. When we compared results in such animals to those in controls, the treated animals exhibited a significantly reduced bacterial load in the blood and other mouse organs, as well as various levels of cytokines/chemokines. Importantly, neutrophil numbers were significantly elevated in the blood of C 6 -HSL-treated mice compared to those in animals given phosphate-buffered saline and then infected with the bacteria. These findings coincided with the fact that neutropenic animals were more susceptible to A. hydrophila infection than normal mice. Our data suggested that neutrophils quickly cleared bacteria by either phagocytosis or possibly another mechanism(s) during infection. In a parallel study, we indeed showed that other predominant immune cells inflicted during A. hydrophila infections, such as murine macrophages, when they were pretreated with AHLs, rapidly phagocytosed bacteria, whereas untreated cells phagocytosed fewer bacteria. This study is the first to report that AHL pretreatment modulates the innate immune response in mice and enhances their survivability during A. hydrophila infection.

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Section: Vol 79 2011 Homoserine Lactones Enhance Innate Immunity Inmentioning

confidence: 92%

Immunomodulatory and Protective Roles of Quorum-Sensing Signaling Molecules N -Acyl Homoserine Lactones during Infection of Mice with Aeromonas hydrophila

et al. 2011

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“…Phagocytosis was measured as described previously [18, 19]. Staphylococcus aureus 1.2386 and Streptococcus pneumoniae 1.8722 were obtained from the China General Microbiological Culture Collection Center (Beijing, China) and fluorescently labeled using Alexa Fluor 488 dye (Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

Exposure to ambient particulate matter alters the microbial composition and induces immune changes in rat lung

Liao

et al. 2017

Respir Res

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BackgroundAmbient particulate matter exposure has been shown to increase the risks of respiratory diseases. However, the role of the lung microbiome and the immune response to inhaled particulate matter are largely unexplored. We studied the influence of biomass fuel and motor vehicle exhaust particles on the lung microbiome and pulmonary immunologic homeostasis in rats.MethodsFifty-seven Sprague–Dawley rats were randomly divided into clean air (CON), biomass fuel (BMF), and motor vehicle exhaust (MVE) groups. After a 4-week exposure, the microbial composition of the lung was assessed by 16S rRNA pyrosequencing, the structure of the lung tissue was assessed with histological analysis, the phagocytic response of alveolar macrophages to bacteria was determined by flow cytometry, and immunoglobulin concentrations were measured with commercial ELISA kits.ResultsThere was no significant difference in lung morphology between the groups. However, the BMF and MVE groups displayed greater bacterial abundance and diversity. Proteobacteria were present in higher proportions in the MVE group, and 12 bacterial families differed in their relative abundances between the three groups. In addition, particulate matter exposure significantly increased the capacity of alveolar macrophages to phagocytose bacteria and induced changes in immunoglobulin levels.ConclusionWe demonstrated that particulate matter exposure can alter the microbial composition and change the pulmonary immunologic homeostasis in the rat lung.

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“…Samples were then analyzed for the formation of bacterial aggregates by flow cytometry as well as confocal microscopy. Flow cytometry forward and side scatter parameters were used to assess bacterial aggregation as previously described [22]. …”

Section: Methodsmentioning

confidence: 99%

PcpA of Streptococcus pneumoniae mediates adherence to nasopharyngeal and lung epithelial cells and elicits functional antibodies in humans

Khan

Sharma

Filkins

et al. 2012

Microbes and Infection

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A flow cytometric assay to quantify in vivo bacterial uptake by alveolar macrophages

Cited by 7 publications

References 11 publications

Immunomodulatory and Protective Roles of Quorum-Sensing Signaling Molecules N -Acyl Homoserine Lactones during Infection of Mice with Aeromonas hydrophila

Immunomodulatory and Protective Roles of Quorum-Sensing Signaling Molecules N -Acyl Homoserine Lactones during Infection of Mice with Aeromonas hydrophila

Exposure to ambient particulate matter alters the microbial composition and induces immune changes in rat lung

PcpA of Streptococcus pneumoniae mediates adherence to nasopharyngeal and lung epithelial cells and elicits functional antibodies in humans

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