title: Clostridium difficile-derived membrane vesicles induce the expression of pro-inflammatory cytokine genes and cytotoxicity in colonic epithelial cells in vitro.
abstract: Clostridium difficile is the most common etiological agent of antibiotic-associated diarrhea in hospitalized and non-hospitalized patients. This study investigated the secretion of membrane vesicles (MVs) from C. difficile and determined the expression of pro-inflammatory cytokine genes and cytotoxicity of C. difficile MVs in epithelial cells in vitro. C. difficile ATCC 43255 and two clinical isolates secreted spherical MVs during in vitro culture. Proteomic analysis revealed that MVs of C. difficile ATCC 43255 contained a total of 262 proteins. Translation-associated proteins were the most commonly identified in C. difficile MVs, whereas TcdA and TcdB toxins were not detected. C. difficile ATCC 43255-derived MVs stimulated the expression of pro-inflammatory cytokine genes, including interleukin (IL)-1β, IL-6, IL-8, and monocyte chemoattractant protein-1 in human colorectal epithelial Caco-2 cells. Moreover, these extracellular vesicles induced cytotoxicity in Caco-2 cells. In conclusion, C. difficile MVs are important nanocomplexes that elicit a pro-inflammatory response and induce cytotoxicity in colonic epithelial cells, which may contribute, along with toxins, to intestinal mucosal injury during C. difficile infection.
authors.affiliation: Department of Microbiology, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
authors.forename: Asiimwe
authors.initials: A
authors.lastname: Nicholas
authors.affiliation: Department of Microbiology, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
authors.forename: Hyejin
authors.initials: H
authors.lastname: Jeon
authors.affiliation: Department of Microbiology, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
authors.forename: Gati Noble
authors.initials: GN
authors.lastname: Selasi
authors.affiliation: Department of Microbiology, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
authors.forename: Seok Hyeon
authors.initials: SH
authors.lastname: Na
authors.affiliation: Department of Microbiology, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
authors.forename: Hyo Il
authors.initials: HI
authors.lastname: Kwon
authors.affiliation: Department of Microbiology, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
authors.forename: Yoo Jeong
authors.initials: YJ
authors.lastname: Kim
authors.affiliation: Drug & Disease Target Team, Korea Basic Science Institute, Daejeon, Republic of Korea.
authors.forename: Chi Won
authors.initials: CW
authors.lastname: Choi
authors.affiliation: Drug & Disease Target Team, Korea Basic Science Institute, Daejeon, Republic of Korea.
authors.forename: Seung Il
authors.initials: SI
authors.lastname: Kim
authors.affiliation: Department of Microbiology, Kyungpook National University School of Medicine, Daegu, Republic of Korea. Electronic address: leejc@knu.ac.kr.
authors.forename: Je Chul
authors.initials: JC
authors.lastname: Lee
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*; D001427:Bacterial Toxins / Q000633:toxicity*; D018938:Caco-2 Cells / Q000187:drug effects; D018929:Cell Culture Techniques; D018932:Chemokine CCL2 / Q000187:drug effects / Q000235:genetics; D016360:Clostridioides difficile / Q000235:genetics / Q000378:metabolism*; D003106:Colon / Q000473:pathology*; D016207:Cytokines / Q000187:drug effects* / Q000235:genetics*; D004761:Enterocolitis, Pseudomembranous / Q000378:metabolism; D004768:Enterotoxins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*; D004847:Epithelial Cells / Q000187:drug effects*; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D056945:Hep G2 Cells / Q000187:drug effects; D006801:Humans; D053583:Interleukin-1beta / Q000187:drug effects / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D016209:Interleukin-8 / Q000187:drug effects / Q000235:genetics; D007413:Intestinal Mucosa / Q000187:drug effects; D046529:Microscopy, Electron, Transmission; D040901:Proteomics
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; C111835:CCL2 protein, human; D018932:Chemokine CCL2; D016207:Cytokines; D004768:Enterotoxins; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile
keywords: C. difficile; Cytotoxicity; Membrane vesicle; Pro-inflammatory cytokines