In vitro cytotoxicity induced by Clostridium perfringens isolate carrying a chromosomal cpe gene is exclusively dependent on sporulation and enterotoxin production.
Clostridium perfringens type A is a common source of food poisoning (FP) and non-food-borne (NFB) gastrointestinal diseases in humans. In the intestinal tract, the vegetative cells sporulate and produce a major pathogenic factor, C. perfringens enterotoxin (CPE). Most type A FP isolates carry a chromosomal cpe gene, whereas NFB type A isolates typically carry a plasmid-encoded cpe. In vitro, the purified CPE protein binds to a receptor and forms pores, exerting a cytotoxic activity in epithelial cells. However, it remains unclear if CPE is indispensable for C. perfringens cytotoxicity. In this study, we examined the cytotoxicity of cpe-harboring C. perfringens isolates co-cultured with human intestinal epithelial Caco-2 cells. The FP strains showed severe cytotoxicity during sporulation and CPE production, but not during vegetative cell growth. While Caco-2 cells were intact during co-culturing with cpe-null mutant derivative of strain SM101 (a FP strain carrying a chromosomal cpe gene), the wild-type level cytotoxicity was observed with cpe-complemented strain. In contrast, both wild-type and cpe-null mutant derivative of the NFB strain F4969 induced Caco-2 cell death during both vegetative and sporulation growth. Collectively, the Caco-2 cell cytotoxicity caused by C. perfringens strain SM101 is considered to be exclusively dependent on CPE production, whereas some additional toxins should be involved in F4969-mediated in vitro cytotoxicity.
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 5988531, Japan. Electronic address: shishimaru@vet.osakafu-u.ac.jp.
Mayo
M
Yasugi
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 5988531, Japan.
Yuki
Y
Sugahara
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 5988531, Japan.
Hidenobu
H
Hoshi
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 5988531, Japan.
Kaori
K
Kondo
Department of Biomedical Sciences, Oregon State University, Dryden Hall 216, Corvallis, OR 97331, USA; Department of Microbiology, Oregon State University, Dryden Hall 216, Corvallis, OR 97331, USA.
Prabhat K
PK
Talukdar
Department of Biomedical Sciences, Oregon State University, Dryden Hall 216, Corvallis, OR 97331, USA; Department of Microbiology, Oregon State University, Dryden Hall 216, Corvallis, OR 97331, USA.
Mahfuzur R
MR
Sarker
School of Marine Science and Technology, Tokai University, 3-20-1 Orido, Shimizu, Shizuoka 4248610, Japan.
Shigeki
S
Yamamoto
Department of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 0208550, Japan.
Yoichi
Y
Kamata
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 5988531, Japan.
Masami
M
Miyake
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D018938:Caco-2 Cells; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003016:Clostridium perfringens / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity*; D004768:Enterotoxins / Q000096:biosynthesis / Q000235:genetics / Q000633:toxicity*; D005517:Foodborne Diseases / Q000382:microbiology*; D005738:Gas Gangrene / Q000382:microbiology*; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D013171:Spores, Bacterial / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism / Q000472:pathogenicity; D014774:Virulence
D001426:Bacterial Proteins; D004768:Enterotoxins
Caco-2 cells; Clostridium perfringens; Cytotoxicity; Enterotoxin; Food poisoning