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Cytochalasin D inhibits lipopolysaccharide-induced tumor necrosis factor production in macrophages
H Shinji1, K S Akagawa, T Yoshida
1Tokyo Institute for Immunopharmacology Inc., Japan.
Abstract:
We reported previously that a reorganization of microfilaments can be observed when macrophages are stimulated with lipopolysaccharide (LPS). This reorganization is not detected with current methods in macrophages of an LPS-nonresponsive C3H/HeJ mouse. These results suggest that the observed microfilament response might be involved in a macrophage-activating process induced by LPS. To investigate this, we studied the effect of cytochalasin D (CD), which inhibits reorganization of microfilaments, on LPS-induced tumor necrosis factor (TNF) production. A concentration of CD incapable of affecting filamentous actin by itself was used in these experiments. When this concentration of CD was added after LPS stimulation, microfilament reorganization and TNF production were inhibited. The suppressive effect of CD on TNF production was confirmed by the observation that TNF-alpha mRNA expression was also inhibited by CD. This inhibitory effect of CD was not specific to TNF, because the production of interleukin-1 and prostaglandin E2 were also inhibited. These effects of CD were observed only when CD was added within the first 20 min after LPS stimulation. These results suggest that the CD-sensitive microfilament response is essential in the signaling pathway for the production of certain monokines in LPS-stimulated macrophages.
Insights
Lipopolysaccharide (LPS) triggers microfilament reorganization in macrophages, crucial for producing tumor necrosis factor (TNF) and other inflammatory mediators. Inhibiting this response with cytochalasin D blocks LPS-induced inflammatory cytokine production.
Area of Science:
- Immunology
- Cell Biology
Background:
- Macrophages play a key role in the immune response.
- Lipopolysaccharide (LPS) is a potent activator of macrophages.
- Microfilament reorganization is an early event following LPS stimulation.
Purpose of the Study:
- To investigate the role of LPS-induced microfilament reorganization in macrophage activation.
- To determine if inhibiting microfilament reorganization affects the production of inflammatory mediators.
Main Methods:
- Macrophages were stimulated with LPS.
- Cytochalasin D (CD), a microfilament inhibitor, was used at a sub-effect concentration.
- Production of TNF, IL-1, and PGE2, as well as TNF-alpha mRNA expression, were measured.
Main Results:
- CD inhibited LPS-induced microfilament reorganization.
- CD suppressed the production of TNF, IL-1, and PGE2.
- Inhibition of cytokine production by CD required addition within 20 minutes of LPS stimulation.
- CD also inhibited TNF-alpha mRNA expression.
Conclusions:
- LPS-induced microfilament reorganization is essential for the signaling pathway leading to the production of certain monokines.
- The actin cytoskeleton plays a critical role in LPS-induced macrophage activation and inflammatory mediator release.
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