Effects of Claudin-1 on the Action of Clostridium perfringens Enterotoxin in Caco-2 Cells
by
Iman Mehdizadeh Gohari 1, Jihong Li 1, Mauricio Navarro 2, Francisco Uzal 2 and Bruce McClane 1,*
Iman Mehdizadeh Gohari 1, Jihong Li 1, Mauricio Navarro 2, Francisco Uzal 2 and Bruce McClane 1,*
1
Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, PA 15219, USA
2
California Animal Health and Food Safety Laboratory System, School of Veterinary Medicine, University of California Davis, San Bernadino, CA 92408, USA
*
Author to whom correspondence should be addressed.
Toxins 2019, 11(10), 582; https://doi.org/10.3390/toxins11100582
Received: 4 September 2019 / Revised: 1 October 2019 / Accepted: 3 October 2019 / Published: 9 October 2019
(This article belongs to the Collection Clostridium difficile/Clostridium sordellii and Clostridium perfringens Toxins)
Clostridium perfringens enterotoxin (CPE) contributes to diarrhea and an often-lethal enterotoxemia. CPE action starts when it binds to claudin receptors, forming a small complex (90 kDa). Six small complexes then oligomerize to create prepores, followed by insertion of beta-hairpins from CPE to form beta-barrel pores named CH-1 or CH-2. Of the ~27 members of the human claudin protein family, only some bind CPE. However, both receptor claudins and the nonreceptor claudin-1 (CLDN-1) are associated with the small and CH-1/CH-2 CPE complexes. Therefore, this study evaluated whether claudin-1 affects CPE action by generating a CLDN-1 null mutant in Caco-2 cells using CRISPR-Cas9. Compared to wild-type Caco-2 cells, paracellular permeability of the CLDN-1 mutant was significantly enhanced, suggesting that claudin-1 may reduce CPE absorption during enterotoxemia. The CLDN-1 mutant was also markedly more sensitive than wild-type Caco-2 cells to apically-applied CPE. The mechanism behind this increased sensitivity involved higher CPE binding by the CLDN-1 mutant vs. wild-type Caco-2 cells, which led to more CH-1/CH-2 complex formation. However, the CH-1/CH-2 complexes formed by the CLDN-1 mutant were less stable or trypsin resistant than those of wild-type cells. These results indicate that, although a nonreceptor, CLDN-1 positively and negatively influences CPE action.
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Keywords:
Clostridium perfringens; enterotoxin; claudins; occludin; tight junctions; diarrheal disease
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MDPI and ACS Style
Mehdizadeh Gohari, I.; Li, J.; Navarro, M.; Uzal, F.; McClane, B. Effects of Claudin-1 on the Action of Clostridium perfringens Enterotoxin in Caco-2 Cells. Toxins 2019, 11, 582.
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