Glucosyltransferase-dependent and independent effects of Clostridioides difficile toxins during infection.
Clostridioides difficile infection (CDI) is the leading cause of nosocomial diarrhea and pseudomembranous colitis in the USA. In addition to these symptoms, patients with CDI can develop severe inflammation and tissue damage, resulting in life-threatening toxic megacolon. CDI is mediated by two larg...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2022-02-01
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| Series: | PLoS Pathogens |
| Online Access: | https://doi.org/10.1371/journal.ppat.1010323 |
| _version_ | 1818272517358354432 |
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| author | F Christopher Peritore-Galve John A Shupe Rory J Cave Kevin O Childress M Kay Washington Sarah A Kuehne D Borden Lacy |
| author_facet | F Christopher Peritore-Galve John A Shupe Rory J Cave Kevin O Childress M Kay Washington Sarah A Kuehne D Borden Lacy |
| author_sort | F Christopher Peritore-Galve |
| collection | DOAJ |
| description | Clostridioides difficile infection (CDI) is the leading cause of nosocomial diarrhea and pseudomembranous colitis in the USA. In addition to these symptoms, patients with CDI can develop severe inflammation and tissue damage, resulting in life-threatening toxic megacolon. CDI is mediated by two large homologous protein toxins, TcdA and TcdB, that bind and hijack receptors to enter host cells where they use glucosyltransferase (GT) enzymes to inactivate Rho family GTPases. GT-dependent intoxication elicits cytopathic changes, cytokine production, and apoptosis. At higher concentrations TcdB induces GT-independent necrosis in cells and tissue by stimulating production of reactive oxygen species via recruitment of the NADPH oxidase complex. Although GT-independent necrosis has been observed in vitro, the relevance of this mechanism during CDI has remained an outstanding question in the field. In this study we generated novel C. difficile toxin mutants in the hypervirulent BI/NAP1/PCR-ribotype 027 R20291 strain to test the hypothesis that GT-independent epithelial damage occurs during CDI. Using the mouse model of CDI, we observed that epithelial damage occurs through a GT-independent process that does not involve immune cell influx. The GT-activity of either toxin was sufficient to cause severe edema and inflammation, yet GT activity of both toxins was necessary to produce severe watery diarrhea. These results demonstrate that both TcdA and TcdB contribute to disease pathogenesis when present. Further, while inactivating GT activity of C. difficile toxins may suppress diarrhea and deleterious GT-dependent immune responses, the potential of severe GT-independent epithelial damage merits consideration when developing toxin-based therapeutics against CDI. |
| first_indexed | 2024-12-12T21:43:20Z |
| format | Article |
| id | doaj.art-53681cccf54a48f9aac596fac3673cfc |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-12-12T21:43:20Z |
| publishDate | 2022-02-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-53681cccf54a48f9aac596fac3673cfc2022-12-22T00:11:00ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742022-02-01182e101032310.1371/journal.ppat.1010323Glucosyltransferase-dependent and independent effects of Clostridioides difficile toxins during infection.F Christopher Peritore-GalveJohn A ShupeRory J CaveKevin O ChildressM Kay WashingtonSarah A KuehneD Borden LacyClostridioides difficile infection (CDI) is the leading cause of nosocomial diarrhea and pseudomembranous colitis in the USA. In addition to these symptoms, patients with CDI can develop severe inflammation and tissue damage, resulting in life-threatening toxic megacolon. CDI is mediated by two large homologous protein toxins, TcdA and TcdB, that bind and hijack receptors to enter host cells where they use glucosyltransferase (GT) enzymes to inactivate Rho family GTPases. GT-dependent intoxication elicits cytopathic changes, cytokine production, and apoptosis. At higher concentrations TcdB induces GT-independent necrosis in cells and tissue by stimulating production of reactive oxygen species via recruitment of the NADPH oxidase complex. Although GT-independent necrosis has been observed in vitro, the relevance of this mechanism during CDI has remained an outstanding question in the field. In this study we generated novel C. difficile toxin mutants in the hypervirulent BI/NAP1/PCR-ribotype 027 R20291 strain to test the hypothesis that GT-independent epithelial damage occurs during CDI. Using the mouse model of CDI, we observed that epithelial damage occurs through a GT-independent process that does not involve immune cell influx. The GT-activity of either toxin was sufficient to cause severe edema and inflammation, yet GT activity of both toxins was necessary to produce severe watery diarrhea. These results demonstrate that both TcdA and TcdB contribute to disease pathogenesis when present. Further, while inactivating GT activity of C. difficile toxins may suppress diarrhea and deleterious GT-dependent immune responses, the potential of severe GT-independent epithelial damage merits consideration when developing toxin-based therapeutics against CDI.https://doi.org/10.1371/journal.ppat.1010323 |
| spellingShingle | F Christopher Peritore-Galve John A Shupe Rory J Cave Kevin O Childress M Kay Washington Sarah A Kuehne D Borden Lacy Glucosyltransferase-dependent and independent effects of Clostridioides difficile toxins during infection. PLoS Pathogens |
| title | Glucosyltransferase-dependent and independent effects of Clostridioides difficile toxins during infection. |
| title_full | Glucosyltransferase-dependent and independent effects of Clostridioides difficile toxins during infection. |
| title_fullStr | Glucosyltransferase-dependent and independent effects of Clostridioides difficile toxins during infection. |
| title_full_unstemmed | Glucosyltransferase-dependent and independent effects of Clostridioides difficile toxins during infection. |
| title_short | Glucosyltransferase-dependent and independent effects of Clostridioides difficile toxins during infection. |
| title_sort | glucosyltransferase dependent and independent effects of clostridioides difficile toxins during infection |
| url | https://doi.org/10.1371/journal.ppat.1010323 |
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