title: Determination of the cytotoxic effect of Clostridium histolyticum culture supernatant on HeLa cells in the presence of protease inhibitors.
abstract: Clostridium histolyticum culture supernatant contains numerous enzymes, which exert a cytotoxic effect on host cells. This includes lethal toxin, clostripain and high-potassium-sensitive toxin. Since the number of C. histolyticum infections increased during the last several years, it seems worthwhile to evaluate whether protease inhibitors, used for the treatment of many diseases, could influence toxicity, and thus, pathogenicity of C. histolyticum. In this study we evaluated in vitro the influence of four common protease inhibitors: aprotinin, phenylmethylsulphonyl fluoride (PMSF), l-1-chloro-3-[4-tosylamido]-7-amino-2-heptanone-HCl (TLCK) and chymostatin on the toxicity of C. histolyticum supernatant towards human epithelial HeLa cells. We show that aprotinin has no effect, while PMSF, TLCK and chymostatin potentiate the cytotoxic activity of C. histolyticum, probably by hindering natural defence mechanisms of cells. In addition, PMSF and TLCK block clostripain enzymatic activity, while chymostatin leaves it intact. Elevated cytotoxicity of the supernatant is not related to the quantity of high-potassium-sensitive toxin, as was reported previously, since desalted supernatant still exerted its strong toxic effect. Our results show that addition of protease inhibitors for treating diseases complicated by concurrent C. histolyticum infection must require special attention.
authors.affiliation: Department of Histology and Embryology, Center of Biostructure Research, Medical University of Warsaw, ul. Chalubinskiego, 502-004 Warsaw, Poland. jjozwiak@atdv.com.pl
authors.forename: Jarosław
authors.initials: J
authors.lastname: Jóźwiak
authors.forename: Aldona
authors.initials: A
authors.lastname: Komar
authors.forename: Ewa
authors.initials: E
authors.lastname: Jankowska
authors.forename: Gayane
authors.initials: G
authors.lastname: Martirosian
mesh_terms: D007611:Aprotinin / Q000494:pharmacology; D001427:Bacterial Toxins / Q000633:toxicity*; D016923:Cell Death / Q000187:drug effects; D047009:Hathewaya histolytica / Q000472:pathogenicity*; D017077:Culture Media, Conditioned; D003546:Cysteine Endopeptidases / Q000633:toxicity; D006367:HeLa Cells; D006801:Humans; D009842:Oligopeptides / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology*; D014105:Tosyl Compounds / Q000494:pharmacology; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology
chemical_list: D001427:Bacterial Toxins; D017077:Culture Media, Conditioned; D009842:Oligopeptides; D011480:Protease Inhibitors; D014105:Tosyl Compounds; D014107:Tosyllysine Chloromethyl Ketone; C003614:4-toluenesulfonyl fluoride; C002101:chymostatin; D007611:Aprotinin; D003546:Cysteine Endopeptidases; C019913:clostripain