Frankowska Natalia, Szarek Klaudia, Iwanicki Adam, Wultańska Dorota, Pituch Hanna, Kabała Monika, Negri Alessandro, Obuchowski Michał, Hinc Krzysztof
Division of Molecular Bacteriology, Medical University of Gdańsk, Gdańsk, Poland.
Intercollegiate Faculty of Biotechnology, University of Gdańsk, Gdańsk, Poland.
Sci Rep. 2025 May 29;15(1):18856. doi: 10.1038/s41598-025-04106-0.
Clostridioides difficile, the causative agent of C. difficile infections (CDI), can be naturally infected by bacterial viruses known as bacteriophages. All characterized bacteriophages of this bacterium are temperate, meaning that upon infection their genetic material integrates and replicates with host's genome. Such lysogenic strains can exhibit altered physiology and virulence, which in turn can be an important factor for epidemiology of CDI. In this study we characterized the phiCDKH02 bacteriophage infecting clinical isolates of C. difficile belonging to hypervirulent ribotypes 027 and 176. The bacteriophage was found to be identical to phi027. To get some insight into the role of this bacteriophage in physiology of its host and interaction with human colon cells, we made use of CRISPR-Cpf1 technology to cure the lysogenic C. difficile of the prophage. The prophage-free strain exhibited altered sporulation efficiency, lowered adhesion and decreased cytopathic effects towards human colon cells associated with decreased production of TcdB. These results emphasize importance of prophages in shaping virulence of C. difficile.
艰难梭菌是艰难梭菌感染（CDI）的病原体，可被称为噬菌体的细菌病毒自然感染。该细菌所有已鉴定的噬菌体都是温和噬菌体，这意味着感染后它们的遗传物质会与宿主基因组整合并复制。这种溶原性菌株可表现出生理和毒力的改变，这反过来可能是CDI流行病学的一个重要因素。在本研究中，我们对感染属于高毒力核糖体分型027和176的艰难梭菌临床分离株的phiCDKH02噬菌体进行了表征。发现该噬菌体与phi027相同。为了深入了解这种噬菌体在其宿主生理学以及与人类结肠细胞相互作用中的作用，我们利用CRISPR-Cpf1技术去除了溶原性艰难梭菌中的前噬菌体。无前噬菌体菌株表现出芽孢形成效率改变、粘附降低以及对人类结肠细胞的细胞病变效应降低，这与TcdB产生减少有关。这些结果强调了前噬菌体在塑造艰难梭菌毒力方面的重要性。
