Clostridium perfringens enterotoxin damages the human intestine in vitro

Abstract

In vitro, Clostridium perfringens enterotoxin (CPE) binds to human ileal epithelium and induces morphological damage concurrently with reduced short-circuit current, transepithelial resistance, and net water absorption. CPE also binds to the human colon in vitro but causes only slight morphological and transport changes that are not statistically significant.

FIG. 1.

Time course effect of CPE on (A) transepithelial net water flux (ΔJ_w), (B) short-circuit current (ΔI_sc), and (C) transepithelial resistance (ΔR_t) in human ileum. After an equilibration period, intestinal mucosal sheets placed in Ussing chambers were incubated at the mucosal side with either Ringer solution alone (solid line) or Ringer solution containing 10 μg/ml of CPE (dotted line) and monitored for 60 min. Each point indicates the mean ± standard error of the mean of four experiments.

FIG. 2.

Morphological effect of CPE on human ileal mucosa. (a) Human ileum after 60 min of incubation with Ringer solution (control) in Ussing chamber. No significant histological abnormalities were observed. Hematoxylin and eosin, 400×. (b) Human ileum treated with CPE in an Ussing chamber for 60 min. Observe the necrosis and desquamation of the superficial epithelium at the tips of the villi. Hematoxylin and eosin, 400×. Bars, 40 μm.

FIG. 3.

Binding of CPE on human intestine. (a) Human ileum treated with CPE in an Ussing chamber for 60 min. Observe positively stained CPE on the apical borders of the enterocytes. Avidin biotin peroxidase, anti-CPE primary antibody, 400×. (b) Human ileum treated with CPE in an Ussing chamber for 60 min. Avidin biotin peroxidase, normal rabbit serum (negative control), 400×. (c) Human colon treated with CPE in an Ussing chamber for 60 min. Observe positively stained CPE on the apical and basolateral borders of the enterocytes. Avidin biotin peroxidase, anti-CPE primary antibody, 400×. (d) Human colon treated with CPE in an Ussing chamber for 60 min. Avidin biotin peroxidase, normal rabbit serum (negative control), 400×. Bars, 40 μm (a and b) and 20 μm (c and d).