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Inhibition of macrophage migration by C. botulinum exoenzyme C3

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Abstract

C3-like exoenzymes are produced by various microorganism including Clostridium botulinum (C3bot), Bacillus cereus and Staphylococcus aureus. C3bot is the prototype of C3-like exoenzymes that specifically ADP-ribosylates and thereby inactivates Rho(A/B/C). C3-like exoenzymes are not yet regarded as virulence factors, as the lack of cell entry domains results in a poor accessibility of the C3-like exoenzymes to cells. In this study, the sensitivity of various cell lines to C3bot has been reinvestigated. Primary monocytes as well as cultured macrophage-like cells including J774A.1 cells and RAW macrophages exhibit a tenfold higher sensitivity to C3bot than fibroblasts and epithelial cells. RhoA ADP-ribosylation by C3bot resulted in the formation of pronounced bipolar protrusions based on defective tail retraction. The formation of bipolar protrusion resulted in inhibited macrophage migration. These findings suggested that macrophages appear to be target cells of C3bot. Migration of macrophage is a prerequiste for their recruitment to the site of pathogen invasion or tissue damage. Inhibition of macrophage migration likely preserves the survival of C3-producing microorganisms. The observations of this study reinforce the paradigm of a role of C3-like exoenzymes as virulence factors.

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Abbreviations

C3bot:

Exoenzyme C3 from C. botulinum

IL-2:

Interleukin-2

PI3K:

Phosphatidylinositide 3′-OH kinase

TcdB:

Toxin B from the C. difficile strain VPI10463

TcdBF:

Toxin B from the C. difficile serotype F strain 1470.

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Acknowledgments

This work was supported by Deutsche Forschungsgemeinschaft (project JU 231/5-1).

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Authors and Affiliations

  1. Institute of Toxicology OE 5340, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Jacqueline Rotsch, Astrid Rohrbeck, Martin May, Tanja Kolbe, Sandra Hagemann, Ilona Schelle, Ingo Just, Harald Genth & Stefanie C. Huelsenbeck

Authors
  1. Jacqueline Rotsch
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  2. Astrid Rohrbeck
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  3. Martin May
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  4. Tanja Kolbe
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  5. Sandra Hagemann
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  6. Ilona Schelle
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  7. Ingo Just
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  8. Harald Genth
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  9. Stefanie C. Huelsenbeck
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Corresponding author

Correspondence to Harald Genth.

Electronic supplementary material

Below is the link to the electronic supplementary material.

J774A.1 macrophage motility was analysed using time lapse microscopy. Images were acquired by time lapse microscopy (DMI6000; Leica) every 10 min for 24 h (MPG 2,582 kb)

J774A.1 macrophages were treated with C3bot (300 nM). Cell motility was analysed using time lapse microscopy. Images were acquired by time lapse microscopy (DMI6000; Leica) every 10 min for 24 h (MPG 2,582 kb)

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Rotsch, J., Rohrbeck, A., May, M. et al. Inhibition of macrophage migration by C. botulinum exoenzyme C3. Naunyn-Schmiedeberg's Arch Pharmacol 385, 883–890 (2012). https://doi.org/10.1007/s00210-012-0764-9

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  • DOI: https://doi.org/10.1007/s00210-012-0764-9

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