ArticleJournal of visualized experiments : JoVE2013
Studying interactions of Staphylococcus aureus with neutrophils by flow cytometry and time lapse microscopy.
Article in Journal of visualized experiments : JoVE, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 25 citations in OpenAlex.
- Agnostic B cell selection approach identifies antibodies against K. pneumoniae that synergistically drive complement activation.Nature communications · 2024Article
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- Human IgG Fc-engineering for enhanced plasma half-life, mucosal distribution and killing of cancer cells and bacteria.Nature communications · 2024Article
- Purified complement C3b triggers phagocytosis and activation of human neutrophils via complement receptor 1.Scientific reports · 2023Article
- Natural Human Immunity Against Staphylococcal Protein A Relies on Effector Functions Triggered by IgG3.Frontiers in immunology · 2022Article
- Population analysis of Legionella pneumophila reveals a basis for resistance to complement-mediated killing.Nature communications · 2021Article
- Staphylococcal protein A inhibits complement activation by interfering with IgG hexamer formation.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Outer membrane permeabilization by the membrane attack complex sensitizes Gram-negative bacteria to antimicrobial proteins in serum and phagocytes.PLoS pathogens · 2021Article
- Use of Flow Cytometry to Evaluate Phagocytosis ofFrontiers in immunology · 2021Article
- Host-specialized fibrinogen-binding by a bacterial surface protein promotes biofilm formation and innate immune evasion.PLoS pathogens · 2019Article
- The Staphylococcus aureus superantigen SElX is a bifunctional toxin that inhibits neutrophil function.PLoS pathogens · 2017Article
- Identification of LukPQ, a novel, equid-adapted leukocidin of Staphylococcus aureus.Scientific reports · 2017Article
- Accessory gene regulator (Agr) functionality in Staphylococcus aureus derived from lower respiratory tract infections.PloS one · 2017Article
- Serine-Aspartate Repeat Protein D Increases Staphylococcus aureus Virulence and Survival in Blood.Infection and immunity · 2017Article
- LukMF' is the major secreted leukocidin of bovine Staphylococcus aureus and is produced in vivo during bovine mastitis.Scientific reports · 2016Article
- Article
- Bright fluorescent Streptococcus pneumoniae for live-cell imaging of host-pathogen interactions.Journal of bacteriology · 2015Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We present methods to study the effect of phenol soluble modulins (PSMs) and other toxins produced and secreted by Staphylococcus aureus on neutrophils. To study the effects of the PSMs on neutrophils we isolate fresh neutrophils using density gradient centrifugation. These neutrophils are loaded with a dye that fluoresces upon calcium mobilization. The activation of neutrophils by PSMs initiates a rapid and transient increase in the free intracellular calcium concentration. In a flow cytometry experiment this rapid mobilization can be measured by monitoring the fluorescence of a pre-loaded dye that reacts to the increased concentration of free Ca(2+). Using this method we can determine the PSM concentration necessary to activate the neutrophil, and measure the effects of specific and general inhibitors of the neutrophil activation. To investigate the expression of the PSMs in the intracellular space, we have constructed reporter fusions of the promoter of the PSMα operon to GFP. When these reporter strains of S. aureus are phagocytosed by neutrophils, the induction of expression can be observed using fluorescence microscopy.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.