Clostridium difficile toxin: Cytoskeletal changes and lactate dehydrogenase release in hepatocytes

Erik M. Grossmann, Walter E. Longo, Donald L. Kaminski, Gregory S. Smith, Colleen E. Murphy, Rodney L. Durham, Marc J. Shapiro, James G. Norman, John E. Mazuski

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Background. We have found that Clostridium difficile toxins can evoke hepatocyte acute-phase protein synthesis, and that this effect is dependent on a functioning interleukin-1 (IL-1) receptor. The present study was undertaken to determine if C. difficile toxicity, as determined by actin rearrangement and lactate dehydrogenase (LDH) release, also requires a functioning IL-1 receptor. Methods. Primary hepatocyte cultures were prepared from normal mice, knockout mice deficient in the IL-1-converting enzyme (ICE), and knockout mice deficient in the IL-1 p80 receptor. Hepatocytes were treated for 24 h with C. difficile culture extract, purified C. difficile toxin A, or purified C. difficile toxin B. The actin cytoskeleton was examined using confocal microscopy, and LDH release was measured by spectrophotometric analysis. Results. C. difficile culture extract, toxin A, and toxin B induced collapse of the actin cytoskeleton in hepatocytes from normal mice. Hepatocytes from both the ICE-deficient mice and the IL-1 p80 receptor-deficient mice demonstrated similar responses to both toxins. These toxins also induced significant LDH release in a concentration-dependent fashion in the normal hepatocytes and the ICE-deficient hepatocytes. However, no significant increase in LDH release was observed in hepatocytes from IL-1 p80 receptor-deficient mice. Conclusions. C. difficile toxins induce actin cytoskeletal collapse independent of IL-1 or the IL-1 receptor. In contrast, toxin-stimulated LDH release was dependent on the presence of the IL-1 receptor. Thus, separate pathways appear to mediate toxic effects as manifested by actin rearrangement and LDH release. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)165-172
Number of pages8
JournalJournal of Surgical Research
Volume88
Issue number2
DOIs
StatePublished - Feb 2000

Keywords

  • Actin
  • Clostridium difficile toxin
  • Cytoskeleton
  • Interleukin-1 receptor

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