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Related Experiment Video

Updated: Jul 11, 2026

Depletion and Reconstitution of Macrophages in Mice
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Clostridioides difficile-Derived Extracellular Vesicles Induce Proinflammatory Responses in Macrophages.

Fangfang Fan1,2, Yu Zeng1,2, Qianli Zhao1

  • 1Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.

Journal of Extracellular Vesicles
|March 29, 2026
PubMed
Summary

Clostridioides difficile extracellular vesicles (EVs) stimulate macrophage inflammation via FliC protein interacting with TLR5. Inhibiting this interaction reduces inflammatory responses, suggesting FliC as a therapeutic target for Clostridioides difficile infection.

Keywords:
Clostridioides difficileFliCextracellular vesiclespro‐inflammatory cytokines

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Area of Science:

  • Microbiology
  • Immunology
  • Cell Biology

Background:

  • Clostridioides difficile (CD) causes significant antibiotic-associated diarrhea.
  • Extracellular vesicles (EVs) are implicated in pathogen-host interactions.

Purpose of the Study:

  • To investigate the role of Clostridioides difficile strain 630-derived EVs (CD630-EVs) in modulating macrophage pro-inflammatory cytokine production.
  • To identify key proteins within CD630-EVs responsible for immune modulation.

Main Methods:

  • Proteomic analysis of CD630-EVs.
  • In vitro stimulation of mouse macrophages with CD630-EVs.
  • Measurement of cytokine production (IL-6, MCP-1, IL-1α, TNF-α).
  • Inhibition studies using a flagellin receptor inhibitor (TH1020) and protein docking analysis.

Main Results:

  • CD630-EVs contain 1064 proteins, including immune-related proteins FliC, TrxA, TrxA2, and HtpG, with FliC being highly expressed.
  • CD630-EVs significantly increased pro-inflammatory cytokine production in macrophages.
  • TH1020 inhibited cytokine production by disrupting the FliC-TLR5 interaction.
  • FliC appears to be a key mediator of CD630-EV-induced inflammation via TLR5 activation.

Conclusions:

  • CD630-EVs modulate macrophage inflammatory responses during Clostridioides difficile infection (CDI).
  • FliC within CD630-EVs is a critical mediator of inflammation, potentially through TLR5 interaction.
  • FliC represents a potential therapeutic target for mitigating intestinal mucosal injury in CDI.