Abstract
This protocol describes a straightforward technique to evaluate the phagocytotic capacity of murine macrophages for Staphylococcus aureus (S. aureus). By staining S. aureus with Hexidium Iodide and staining murine bone marrow-derived macrophages (BMDMs) with FITC, the macrophage bacterial up-taking ability can be rapidly analyzed by flow cytometry. S. aureus is a Gram-positive bacteria causing severe human and animal infections. Host immune cells such as macrophages serve to eliminate S. aureus by phagocytosing the pathogen and save the host from life-threatening diseases. Study of host macrophage ability to phagocytose S. aureus is important for understanding the host-pathogen interaction and can help to elucidate the pathogenesis of S. aureus infection. This protocol may also be applied for macrophage phagocytotic assay of other gram-positive bacteria.
Materials and Reagents
Equipment
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Procedure
Recipes
Acknowledgments
This work was supported by NIH R01-AI068804. Dr. Fowler was supported by Mid Career Mentoring Award K24-AI093969. The fluorescence staining of Staphylococcus aureus was adapted from Mason et al. (1998), and differentiation of bone marrow-derived macrophages was adapted from Weischenfeldt and Porse (2008).
References
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