Identification of an epithelial cell receptor responsible for Clostridium difficile TcdB-induced cytotoxicity
| Autor: | D. Borden Lacy, Donald H. Rubin, Ramyavardhanee Chandrasekaran, Michelle E. LaFrance, Melissa A. Farrow, Jinsong Sheng |
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| Rok vydání: | 2015 |
| Předmět: |
Colon
Bacterial Toxins Nectins Mutagenesis (molecular biology technique) Biology Microbiology Insertional mutagenesis Enterotoxins Bacterial Proteins Humans Secretion Cytotoxicity Analysis of Variance Multidisciplinary Clostridioides difficile Cell adhesion molecule Genetic Complementation Test Antibodies Monoclonal Epithelial Cells Pseudomembranous colitis Biological Sciences Clostridium difficile Mutagenesis Insertional Ectodomain Caco-2 Cells Cell Adhesion Molecules HeLa Cells |
| Zdroj: | Proceedings of the National Academy of Sciences. 112:7073-7078 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1500791112 |
| Popis: | Clostridium difficile is the leading cause of hospital-acquired diarrhea in the United States. The two main virulence factors of C. difficile are the large toxins, TcdA and TcdB, which enter colonic epithelial cells and cause fluid secretion, inflammation, and cell death. Using a gene-trap insertional mutagenesis screen, we identified poliovirus receptor-like 3 (PVRL3) as a cellular factor necessary for TcdB-mediated cytotoxicity. Disruption of PVRL3 expression by gene-trap mutagenesis, shRNA, or CRISPR/Cas9 mutagenesis resulted in resistance of cells to TcdB. Complementation of the gene-trap or CRISPR mutants with PVRL3 resulted in restoration of TcdB-mediated cell death. Purified PVRL3 ectodomain bound to TcdB by pull-down. Pretreatment of cells with a monoclonal antibody against PVRL3 or prebinding TcdB to PVRL3 ectodomain also inhibited cytotoxicity in cell culture. The receptor is highly expressed on the surface epithelium of the human colon and was observed to colocalize with TcdB in both an explant model and in tissue from a patient with pseudomembranous colitis. These data suggest PVRL3 is a physiologically relevant binding partner that can serve as a target for the prevention of TcdB-induced cytotoxicity in C. difficile infection. |
| Databáze: | OpenAIRE |
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