title: Noncytotoxic Clostridium perfringens enterotoxin (CPE) variants localize CPE intestinal binding and demonstrate a relationship between CPE-induced cytotoxicity and enterotoxicity.
abstract: Clostridium perfringens enterotoxin (CPE) causes the symptoms of a very common food poisoning. To assess whether CPE-induced cytotoxicity is necessary for enterotoxicity, a rabbit ileal loop model was used to compare the in vivo effects of native CPE or recombinant CPE (rCPE), both of which are cytotoxic, with those of the noncytotoxic rCPE variants rCPE D48A and rCPE(168-319). Both CPE and rCPE elicited significant fluid accumulation in rabbit ileal loops, along with severe mucosal damage that starts at villus tips and then progressively affects the entire villus, including necrosis of epithelium and lamina propria, villus blunting and fusion, and transmural edema and hemorrhage. Similar treatment of ileal loops with either of the noncytotoxic rCPE variants produced no visible histologic damage or fluid transport changes. Immunohistochemistry revealed strong CPE or rCPE(168-319) binding to villus tips, which correlated with the abundant presence of claudin-4, a known CPE receptor, in this villus region. These results support (i) cytotoxicity being necessary for CPE-induced enterotoxicity, (ii) the CPE sensitivity of villus tips being at least partially attributable to the abundant presence of receptors in this villus region, and (iii) claudin-4 being an important intestinal receptor for CPE. Finally, rCPE(168-319) was able to partially inhibit CPE-induced histologic damage, suggesting that noncytotoxic rCPE variants might be useful for protecting against some intestinal effects of CPE.
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: James G
authors.initials: JG
authors.lastname: Smedley
authors.forename: Juliann
authors.initials: J
authors.lastname: Saputo
authors.forename: Jacquelyn C
authors.initials: JC
authors.lastname: Parker
authors.forename: Mariano E
authors.initials: ME
authors.lastname: Fernandez-Miyakawa
authors.forename: Susan L
authors.initials: SL
authors.lastname: Robertson
authors.forename: Bruce A
authors.initials: BA
authors.lastname: McClane
authors.forename: Francisco A
authors.initials: FA
authors.lastname: Uzal
mesh_terms: D000818:Animals; D002470:Cell Survival; D062506:Claudin-4; D003016:Clostridium perfringens / Q000235:genetics / Q000472:pathogenicity*; D004487:Edema / Q000139:chemically induced; D004768:Enterotoxins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity*; D005260:Female; D006470:Hemorrhage / Q000139:chemically induced; D007082:Ileum / Q000473:pathology; D007413:Intestinal Mucosa / Q000737:chemistry / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000032:analysis; D009336:Necrosis / Q000139:chemically induced; D011485:Protein Binding; D011817:Rabbits
chemical_list: D062506:Claudin-4; D004768:Enterotoxins; D008565:Membrane Proteins; C035598:enterotoxin, Clostridium