Cellular vacuolation and mitochondrial-associated factors induced by Clostridium perfringens epsilon toxin detected using acoustic flow cytometry.
Epsilon toxin (ETX) produced by Clostridium perfringens types B and D is a potent toxin that is responsible for fatal enterotoxaemia. In vitro, ETX, which is considered as a pore-forming toxin, forms a heptamer in Madin-Darby canine kidney (MDCK) cell membranes, which is considered to be a pre-pore stage. After binding of the ETX, vacuoles inside cell cytoplasm are produced. ETX causes decreased levels of essential coenzymes required for host cell energy. Here, we optimized and applied acoustic flow cytometry analysis in order to gain further insight into ETX-pathogenesis. Using acoustic flow cytometer analysis, which considered highly sensitive, ETX-exposed MDCK cells revealed mitochondrial membrane decreases followed by 25.48% and 45.45% of the exposed cells expressing the Bax and BCL-2 proteins at a pre-pore stage, respectively. These results together with high cytotoxicity and visualization of cell vacuoles, demonstrates that acoustic flow cytometry analysis potentially represents an effective tool to study ETX pathogenesis.
University of São Paulo State, Departamento de Apoio, Produção e Saúde Animal, Rua Clóvis Pestana, 793, Araçatuba 16050-680, São Paulo, Brazil.
Marina C
MC
Ferrarezi
Vera C L M
VC
Curci
Tereza C
TC
Cardoso
D000162:Acoustics; D000818:Animals; D001427:Bacterial Toxins / Q000633:toxicity*; D002460:Cell Line; D004285:Dogs; D004847:Epithelial Cells / Q000187:drug effects* / Q000502:physiology*; D005434:Flow Cytometry / Q000379:methods; D008928:Mitochondria / Q000187:drug effects* / Q000502:physiology*; D014617:Vacuoles / Q000187:drug effects* / Q000378:metabolism*
D001427:Bacterial Toxins; C039811:Clostridium perfringens epsilon-toxin
Clostridium perfringens; In vitro assay; Mitochondria; Mitochondrion; Necrosis