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Antimicrobial Agents and Chemotherapy, March 1999, p. 697-698, Vol. 43, No. 3
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Fosfomycin Alters Lipopolysaccharide-Induced
Inflammatory Cytokine Production in Mice
Tetsuya
Matsumoto,1,*
Kazuhiro
Tateda,1
Shuichi
Miyazaki,1
Nobuhiko
Furuya,1
Akira
Ohno,1
Yoshikazu
Ishii,1
Yoichi
Hirakata,2 and
Keizo
Yamaguchi1
Department of Microbiology, Toho University
School of Medicine, Omori-Nishi, Ota-ku, Tokyo,1
and Department of Laboratory Medicine, Nagasaki University
School of Medicine, Sakamoto, Nagasaki,2 Japan
Received 22 June 1998/Returned for modification 25 October
1998/Accepted 26 December 1998
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ABSTRACT |
To determine the mechanisms of immunomodulating action of
fosfomycin (FOF), we examined its effect on the production of
inflammatory cytokines in mice injected with lipopolysaccharide (LPS).
Treatment with FOF significantly lowered the peak serum levels of tumor necrosis factor alpha and interleukin-1
, indicating that FOF alters
inflammatory cytokine production after LPS stimulation.
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TEXT |
Fosfomycin (FOF),
1-cis-1,2-epoxypropylphosphoric acid, is a broad-spectrum
bactericidal antibiotic not structurally related to other classes of
antimicrobial agents. We have recently reported that FOF exerts a
protective effect against murine gut-derived sepsis caused by
Pseudomonas aeruginosa (6). In that study, we
demonstrated that the enantiomer of FOF [FOF(+)], which lacks antimicrobial activity, also exerts a protective effect. FOF(+) significantly suppresses the concentrations of tumor necrosis factor
alpha (TNF-
), interleukin-1 (IL-1), and interleukin-6 (IL-6)
in sera of mice with gut-derived sepsis. Several studies have indicated
that the excessive production of host inflammatory cytokines might be
responsible for the morbidity associated with septic shock (1, 9,
10). Previous studies from our laboratory also demonstrated that
TNF and IL-1 might facilitate bacterial translocation and result in
deterioration of gut-derived sepsis caused by P. aeruginosa
in mice (5, 7). These results suggested that the alteration
of inflammatory cytokine production by FOF and FOF(+) may potentially
affect the pathophysiology of septic shock. However, we could not
demonstrate a direct alternative effect of FOF on cytokine production
in vivo, because murine gut-derived sepsis is a complex model for the
analysis of the protective mechanisms of FOF. Therefore, in this study,
we evaluated the effects of FOF on inflammatory cytokine production
with a simple animal model induced by inoculation with
lipopolysaccharide (LPS).
Specific-pathogen-free male ddY mice (Japan Shizuoka Laboratory Center
Co., Shizuoka, Japan) were or were not treated with 200 mg of FOF
(Meiji Seika, Tokyo, Japan) per kg by intravenous injection. Ten
minutes after FOF or saline injection, 2 mg of LPS (Escherichia
coli O55:B5; Difco Laboratories, Detroit, Mich.) per kg was
injected intraperitoneally. Mice were then sacrificed at the indicated
time intervals (0, 1.5, 3, 6, and 12 h; n = 10 [at each time point]) by inhalation of ether, and cardiac blood samples were immediately obtained. Serum samples were preserved at
80°C until measurements of cytokine concentrations were taken. Concentrations of inflammatory cytokines in serum were determined by
using enzyme-linked immunosorbent assay kits (kits for TNF-, gamma
interferon [IFN-
], and IL-6 were from Endogen Inc., Boston, Mass.,
and that for IL-1 was from Genzyme Corp., Boston, Mass.). Each
experiment was performed twice.
Treatment with FOF significantly lowered the peak concentration of
TNF- in serum, compared with that in saline-treated control mice
(0.56 ± 0.26 versus 1.35 ± 0.33 ng/ml; Fig.
1). The peak level of IL-1 was also
significantly lowered in FOF-treated mice, compared with that in
saline-treated control mice (163.8 ± 34.6 versus 468.6 ± 54.7 pg/ml; Fig. 2). Furthermore, we
determined the concentration of TNF- in serum after treatment of
mice with various doses of FOF and noted a dose-response effect of FOF
on TNF- production (Fig. 3).
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FIG. 1.
Effect of FOF on serum TNF- kinetics after LPS
stimulation in mice. Mice in groups of 10 were each treated
intravenously with 200 mg of FOF per kg (open circles) or with saline
alone (solid circles) 10 min before LPS inoculation. Data are
means ± standard deviations (error bars). §, P < 0.05.
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FIG. 2.
Effect of FOF on serum IL-1 kinetics after LPS
stimulation in mice. Mice in groups of 10 were each treated
intravenously with 200 mg of FOF per kg (open circles) or with saline
alone (solid circles) 10 min before LPS inoculation. Data are
means ± standard deviations (error bars). §, P < 0.05.
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FIG. 3.
Dose-response effect of FOF on TNF- production
following LPS stimulation in mice. Mice in groups of 8 were each
treated intravenously with 0.8, 4, 20, or 100 mg of FOF per kg (white
bars) or with saline alone (gray bar) 10 min before LPS inoculation.
Data are means ± standard deviations (error bars). §,
P < 0.05.
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Although the differences in IFN- and IL-6 concentrations between
FOF-treated and saline-treated mice were rather marginal, FOF
significantly lowered IFN- concentrations (5.51 ± 1.48 versus 0.35 ± 0.35 ng/ml; P < 0.05) and increased IL-6
concentrations (24.9 ± 1.5 versus 20.4 ± 1.6 ng/ml;
P < 0.05) in the initial stage (the first 1.5 h),
compared to those in saline-treated control mice. Because there were
only weak influences on IFN- and IL-6, we speculate that the effects
of FOF on changes in inflammatory cytokines are mediated largely
through TNF- and IL-1.
Concerning the FOF treatment schedule, there were no significant
differences in cytokine levels between groups treated with FOF 10 min
or 1 day before LPS administration. On the other hand, FOF treatment
1 h after LPS administration did not show a significant effect,
compared to saline treatment on the same schedule (data not shown).
With regard to the route of treatment, there were no significant
differences between groups treated intravenously and those treated
intraperitoneally (data not shown).
In a series of preliminary studies, we confirmed that the FOF
preparation used in the present study contained an amount of endotoxin
below the detection level and that the administration of FOF alone did
not produce a rise in concentrations of TNF-, IL-1, IL-6, and
IFN- in mouse sera (data not shown). We also confirmed that all mice
survived more than 14 days after treatment with 2 mg of LPS per kg, the
dose used in the present study.
Morikawa et al. (8) reported the effects of FOF on cytokine
synthesis by LPS-stimulated human monocytes in vitro. An interesting finding in their report was that FOF decreased the rate of synthesis of
TNF and IL-1 but increased that of the synthesis of IL-6. Our results
in the present study were consistent with these findings. However, the
enhancement of IL-6 production in this study was small, and we could
not directly demonstrate whether this enhancement influenced the
expression of other cytokines.
Although the exact reasons for the contradictory result in a recent
study (6) concerning the effect of FOF on IL-6 production are not known at present, we speculate that some possible reasons for
these contradictory results might include the pathophysiological differences between endotoxin shock and septic shock. For example, not
only LPS but also various types of exotoxins may influence the
pathophysiology of septic shock (2-4).
Our results indicate that FOF directly alters the production of
inflammatory cytokines, especially TNF- and IL-1, in vivo. Further studies are necessary to investigate the mechanisms of this
effect. We are currently investigating the effects of FOF on the
expression of LPS-induced cellular surface molecules.
| |
ACKNOWLEDGMENTS |
We are grateful to Shogo Kuwahara for useful advice and to Yasuko
Kaneko for expert technical assistance. We also thank F. G. Issa
(Word-Medex, Sydney, Australia) for careful reading and editing of the manuscript.
This work was supported by a research grant provided by The Japan
Health Sciences Foundation, Tokyo, Japan.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Department of
Microbiology, Toho University School of Medicine, 5-21-16 Omori-Nishi, Ota-ku, Tokyo 143-8540, Japan. Phone: 81 (3) 3762-4151, ext. 2396. Fax:
81 (3) 5493-5415. E-mail: tetsu{at}med.toho-u.ac.jp.
| |
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Antimicrobial Agents and Chemotherapy, March 1999, p. 697-698, Vol. 43, No. 3
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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