Clostridium perfringens Delta-Toxin Damages the Mouse Small Intestine.

Soshi Seike,Masaya Takehara,Masahiro Nagahama,Keiko Kobayashi

doi:10.3390/toxins11040232

Soshi Seike, Masaya Takehara + Show 2 more

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https://doi.org/10.3390/toxins11040232

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Abstract

Clostridium perfringens strains B and C cause fatal intestinal diseases in animals. The secreted pore-forming toxin delta-toxin is one of the virulence factors of the strains, but the mechanism of intestinal pathogenesis is unclear. Here, we investigated the effects of delta-toxin on the mouse ileal loop. Delta-toxin caused fluid accumulation and intestinal permeability to fluorescein isothiocyanate (FITC)-dextran in the mouse ileal loop in a dose- and time-dependent manner. Treatment with delta-toxin induced significant histological damage and shortening of villi. Delta-toxin activates a disintegrin and metalloprotease (ADAM) 10, leading to the cleavage of E-cadherin, the epithelial adherens junction protein, in human intestinal epithelial Caco-2 cells. In this study, E-cadherin immunostaining in mouse intestinal epithelial cells was almost undetectable 1 h after toxin treatment. ADAM10 inhibitor (GI254023X) blocked the toxin-induced fluid accumulation and E-cadherin loss in the mouse ileal loop. Delta-toxin stimulated the shedding of intestinal epithelial cells. The shedding cells showed the accumulation of E-cadherin in intracellular vesicles and the increased expression of active caspase-3. Our findings demonstrate that delta-toxin causes intestinal epithelial cell damage through the loss of E-cadherin cleaved by ADAM10.

Highlights

Delta-toxin is a β-pore-forming-toxin (β-PFT) produced by Clostridium perfringens strains B and C [1]
To confirm that delta-toxin actively caused intestinal damage, we examined the effect of the preincubation of delta-toxin with an anti-delta-toxin antiserum
It has been reported that delta-toxin disrupts the barrier function of Caco-2 cells, as evidenced by reduced transepithelial electrical resistance (TEER) and decreased cellular levels of adherence junction protein

Summary

Introduction

Delta-toxin is a β-pore-forming-toxin (β-PFT) produced by Clostridium perfringens strains B and C [1]. Delta-toxin hemolyzes the red blood cells of pigs, goats, and sheep [1,5,6]. The toxin exhibits cytotoxic activity against multiple cell types, including macrophages, monocytes, and platelets from various animal species [1,7,8]. Delta-toxin has been assigned to the β-PFT family, which includes alpha-toxin from Staphylococcus aureus and beta-toxin and NetB toxin from C. perfringens [9,10]. The structure of delta-toxin resembles alpha-toxin and NetB toxin [11]. Delta-toxin forms a mushroom-shaped heptameric pore similar to that of alpha-toxin from S. aureus [11]

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