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Ingested spores germinate in the intestine establishing a population of cells that produce toxins and spores. The pathogenicity locus, PaLoc, comprises several genes, including those coding for TcdA\/B, for the holin-like TcdE protein, and for TcdR, an auto-regulatory RNA polymerase sigma factor essential for <jats:italic>tcdA<\/jats:italic>\/<jats:italic>B<\/jats:italic> and <jats:italic>tcdE<\/jats:italic> expression. Here we show that <jats:italic>tcdR<\/jats:italic>, <jats:italic>tcdA<\/jats:italic>, <jats:italic>tcdB<\/jats:italic> and <jats:italic>tcdE<\/jats:italic> are expressed in a fraction of the sporulating cells, in either the whole sporangium or in the forespore. The whole sporangium pattern is due to protracted expression initiated in vegetative cells by \u03c3<jats:sup>D<\/jats:sup>, which primes the TcdR auto-regulatory loop. In contrast, the forespore-specific regulatory proteins \u03c3<jats:sup>G<\/jats:sup> and SpoVT control TcdR production and <jats:italic>tcdA\/tcdB<\/jats:italic> and <jats:italic>tcdE<\/jats:italic> expression in this cell. We detected TcdA at the spore surface, and we show that wild type and \u0394<jats:italic>tcdA<\/jats:italic> or \u0394<jats:italic>tcdB<\/jats:italic> spores but not \u0394<jats:italic>tcdR<\/jats:italic> or \u0394<jats:italic>tcdA\/<\/jats:italic>\u0394<jats:italic>tcdB<\/jats:italic> spores are cytopathic against HT29 and Vero cells, indicating that spores may serve as toxin-delivery vehicles. Since the addition of TcdA and TcdB enhance binding of spores to epithelial cells, this effect may occur independently of toxin production by vegetative cells.<\/jats:p>","DOI":"10.1038\/s42003-024-06521-x","type":"journal-article","created":{"date-parts":[[2024,7,10]],"date-time":"2024-07-10T12:02:17Z","timestamp":1720612937000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Spores of Clostridioides difficile are toxin delivery vehicles"],"prefix":"10.1038","volume":"7","author":[{"given":"Carolina P.","family":"Cassona","sequence":"first","affiliation":[]},{"given":"Sara","family":"Ramalhete","sequence":"additional","affiliation":[]},{"given":"Khira","family":"Amara","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4686-2533","authenticated-orcid":false,"given":"Thomas","family":"Candela","sequence":"additional","affiliation":[]},{"given":"Imad","family":"Kansau","sequence":"additional","affiliation":[]},{"given":"C\u00e9cile","family":"Den\u00e8ve-Larrazet","sequence":"additional","affiliation":[]},{"given":"Claire","family":"Janoir-Jouveshomme","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5592-8397","authenticated-orcid":false,"given":"Lu\u00eds Jaime","family":"Mota","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7176-8453","authenticated-orcid":false,"given":"Bruno","family":"Dupuy","sequence":"additional","affiliation":[]},{"given":"M\u00f3nica","family":"Serrano","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4292-4802","authenticated-orcid":false,"given":"Adriano O.","family":"Henriques","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2024,7,10]]},"reference":[{"key":"6521_CR1","doi-asserted-by":"publisher","first-page":"609","DOI":"10.1038\/nrmicro.2016.108","volume":"14","author":"MC Abt","year":"2016","unstructured":"Abt, M. 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