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Updated: Feb 6, 2026

Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds
Published on: September 8, 2021
Intracellular activity of antimicrobial compounds used for Staphylococcus aureus nasal decolonization
J Rigaill1,2, M F Morgene2, M Gavid3
1Group for Mucosal Immunity and Pathogen Agents (GIMAP), EA 3064, University of Lyon, St-Etienne, France.
Background:
Staphylococcus aureus is able to invade mammalian cells during infection and was recently observed inside nasal mucosa of healthy carriers.
Objectives:
To determine the intracellular activity of antimicrobial compounds used for decolonization procedures using a cell model mimicking S. aureus nasal epithelium invasion.
Patients And Methods:
HaCaT cells and human nasal epithelial cells (HNECs) recovered from nasal swabs of S. aureus carriers were visualized by confocal laser scanning microscopy to detect intracellular S. aureus cells. An HaCaT cell model, mimicking S. aureus internalization observed ex vivo in HNECs, was used to assess the intracellular activity against S. aureus of 21 antimicrobial compounds used for nasal decolonization, including mupirocin and chlorhexidine.
Results:
HaCaT cells and HNECs were found to internalize S. aureus with the same focal pattern. Most antimicrobial compounds tested on HaCaT cells were shown to have weak activity against intracellular S. aureus. Some systemic antimicrobials, including fusidic acid, clindamycin, linezolid, minocycline, ciprofloxacin, moxifloxacin, rifampicin and levofloxacin, reduced S. aureus intracellular loads by 0.43-1.66 log cfu/106 cells compared with the control (P < 0.001). By contrast, mupirocin and chlorhexidine reduced the S. aureus intracellular load by 0.19 and 0.23 log cfu/106 cells, respectively.
Conclusions:
These data indicate that most of the antimicrobial compounds used for nasal decolonization, including mupirocin and chlorhexidine, exhibit weak activity against intracellular S. aureus using the HaCaT cell model. This work emphasizes the need to better understand the role of the S. aureus intracellular reservoir during nasal colonization in order to improve decolonization procedures.
Insights
Most antimicrobial nasal decolonization compounds show weak activity against intracellular Staphylococcus aureus. Systemic antimicrobials were more effective than topical agents like mupirocin and chlorhexidine in reducing bacterial load within cells.
Area of Science:
- Microbiology
- Infectious Diseases
- Pharmacology
Background:
- Staphylococcus aureus invades mammalian cells during infection.
- Intracellular S. aureus has been detected in the nasal mucosa of healthy carriers.
- The role of intracellular bacteria in nasal colonization is increasingly recognized.
Purpose of the Study:
- To evaluate the efficacy of antimicrobial compounds against intracellular S. aureus.
- To assess the activity of nasal decolonization agents in a relevant cell model.
- To compare the effectiveness of different antimicrobial classes against intracellular bacteria.
Main Methods:
- Developed a HaCaT cell model mimicking S. aureus nasal epithelium invasion.
- Utilized confocal laser scanning microscopy to detect intracellular S. aureus.
- Tested 21 antimicrobial compounds, including mupirocin and chlorhexidine, for intracellular activity.
Main Results:
- Both HaCaT cells and human nasal epithelial cells (HNECs) internalized S. aureus.
- Most tested antimicrobial compounds demonstrated limited activity against intracellular S. aureus.
- Systemic antimicrobials (e.g., fluoroquinolones, clindamycin) significantly reduced intracellular S. aureus loads (0.43-1.66 log cfu/10^6 cells).
- Mupirocin and chlorhexidine showed minimal reduction in intracellular S. aureus (0.19 and 0.23 log cfu/10^6 cells, respectively).
Conclusions:
- Current nasal decolonization agents, including mupirocin and chlorhexidine, have poor efficacy against intracellular S. aureus.
- Systemic antimicrobials show greater potential for targeting intracellular S. aureus reservoirs.
- Further research is needed to understand the intracellular reservoir's role in nasal colonization and to develop improved decolonization strategies.
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