The Sialidase NanS Enhances Non-TcsL Mediated Cytotoxicity of Clostridium sordellii.
The clostridia produce an arsenal of toxins to facilitate their survival within the host environment. TcsL is one of two major toxins produced by Clostridium sordellii, a human and animal pathogen, and is essential for disease pathogenesis of this bacterium. C. sordellii produces many other toxins, but the role that they play in disease is not known, although previous work has suggested that the sialidase enzyme NanS may be involved in the characteristic leukemoid reaction that occurs during severe disease. In this study we investigated the role of NanS in C. sordellii disease pathogenesis. We constructed a nanS mutant and showed that NanS is the only sialidase produced from C. sordellii strain ATCC9714 since sialidase activity could not be detected from the nanS mutant. Complementation with the wild-type gene restored sialidase production to the nanS mutant strain. Cytotoxicity assays using sialidase-enriched culture supernatants applied to gut (Caco2), vaginal (VK2), and cervical cell lines (End1/E6E7 and Ect1/E6E7) showed that NanS was not cytotoxic to these cells. However, the cytotoxic capacity of a toxin-enriched supernatant to the vaginal and cervical cell lines was substantially enhanced in the presence of NanS. TcsL was not the mediator of the observed cytotoxicity since supernatants harvested from a TcsL-deficient strain displayed similar cytotoxicity levels to TcsL-containing supernatants. This study suggests that NanS works synergistically with an unknown toxin or toxins to exacerbate C. sordellii-mediated tissue damage in the host.
Infection and Immunity Program, Biomedicine Discovery Institute and Department of Microbiology, Monash University, Clayton 3800, Australia. milena.awad@monash.edu.
Milena M
MM
Awad
Infection and Immunity Program, Biomedicine Discovery Institute and Department of Microbiology, Monash University, Clayton 3800, Australia. julie.singleton@monash.edu.
Julie
J
Singleton
Infection and Immunity Program, Biomedicine Discovery Institute and Department of Microbiology, Monash University, Clayton 3800, Australia. dena.lyras@monash.edu.
Dena
D
Lyras
D001426:Bacterial Proteins / Q000235:genetics / Q000633:toxicity*; D001427:Bacterial Toxins / Q000235:genetics* / Q000633:toxicity; D018938:Caco-2 Cells; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D047989:Paraclostridium sordellii / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D009154:Mutation; D009439:Neuraminidase / Q000235:genetics / Q000633:toxicity*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D009439:Neuraminidase
Clostridium sordellii; clostridia; lethal toxin; pathogenesis; sialidase; toxins