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Staphylococcus aureus phagocytosis. A new cytofluorometric method using FITC and paraformaldehyde
B Cantinieaux1, C Hariga, P Courtoy
1Department of Haematology, University Hospital St. Pierre, Brussels, Belgium.
Journal of Immunological Methods
|July 26, 1989
Summary
A novel flow cytometry (FCM) method accurately quantifies Staphylococcus aureus uptake by human neutrophils. This reproducible technique distinguishes adherent from ingested bacteria, enabling sensitive detection of phagocytosis.
Area of Science:
- Immunology
- Microbiology
- Biotechnology
Background:
- Phagocytosis by human polymorphonuclear neutrophils (PMN) is crucial for controlling bacterial infections.
- Accurate quantification of bacterial uptake by immune cells is essential for understanding host-pathogen interactions.
Purpose of the Study:
- To develop and validate a new flow cytometry (FCM) method for measuring Staphylococcus aureus uptake by human PMN.
- To provide a reproducible and sensitive assay for phagocytosis studies.
Main Methods:
- Bacteria (Staphylococcus aureus) were labeled with fluorescein isothiocyanate (FITC).
- Human PMN were incubated with labeled bacteria in a suspension assay.
- Phagocytosis was quantified using FCM, with crystal violet differentiating adherent and ingested bacteria.
Main Results:
- The FCM method showed good agreement with a reference microscopic technique for quantifying phagocytic indices.
- The assay demonstrated reproducibility and the ability to detect low phagocyte values.
- FCM readings could be delayed without significantly affecting results.
Conclusions:
- The described FCM method offers a reliable, reproducible, and sensitive approach for measuring Staphylococcus aureus phagocytosis by human PMN.
- This technique facilitates the study of immune cell function and bacterial clearance.