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1 Second Dept of Internal Medicine, and 2 Dept of Pharmacotherapeutics, Nagasaki University Graduate School of Pharmaceutical Sciences, and 3 Division of Molecular & Clinical Microbiology, Dept of Molecular Microbiology & Immunology, Nagasaki University Graduate School of Medical Sciences, Nagasaki, Japan.
CORRESPONDENCE: K. Yanagihara, Second Dept of Internal Medicine, Nagasaki University School of Medicine, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan. Fax: 81 958497285. E-mail: kyana-ngs{at}umin.ac.jp
Keywords: Azithromycin, clarithromycin, macrolide-resistant Streptococcus pneumoniae, pneumolysin, sub-minimum inhibitory concentrations
Received: October 6, 2005
Accepted January 10, 2006
To clarify the discrepancy between increasing resistance and conservative clinical effects of macrolides on macrolide-resistant Streptococcus pneumoniae, the authors evaluated the effects of sub-minimum inhibitory concentrations of macrolides on pneumolysin.
In vitro, S. pneumoniae was incubated with 1, 2 and 4 µg·mL–1 of clarithromycin (CLR) and azithromycin (AZM) for 8 h. Western blot analysis and haemolytic assay were performed to examine the production and activities of pneumolysin. In vivo, mice were infected with S. pneumoniae intra-nasally and treated with CLR (40 or 200 mg·kg–1 twice daily) or AZM (40 or 200 mg·kg–1 once daily) orally for 7 days. After 72 h post-infection, western blot analysis was performed to examine pneumolysin production in lungs. Survival rates were observed for 10 days.
In vitro, every concentration of macrolide inhibited pneumolysin production more than the control. CLR (2 and 4 µg·mL–1) and AZM (4 µg·mL–1) reduced the pneumolysin activities more than the control. In vivo, macrolides (200 mg·kg–1) reduced pneumolysin in murine lungs more than the control. CLR (40 and 200 mg·kg–1) and AZM (200 mg·kg–1) improved the survival rates more than the control.
The study results show that sub-minimum inhibitory concentrations of macrolides reduced pneumolysin. This might be related to the effectiveness of macrolides against pneumonia caused by high-level macrolide-resistant Streptococcus pneumoniae. Further investigations are necessary to evaluate the effects of macrolides on macrolide-resistant Streptococcus pneumoniae.
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