Membrane Cholesterol Is Crucial for Clostridium difficile Surface Layer Protein Binding and Triggering Inflammasome Activation
Clostridium difficile, an obligate anaerobic gram-positive bacillus, generates spores and is commonly found colonizing the human gut. Patients with C. difficile infection (CDI) often exhibit clinical manifestations of pseudomembranous colitis or antibiotic-associated diarrhea. Surface layer proteins...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-07-01
|
| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2020.01675/full |
| _version_ | 1818509535906627584 |
|---|---|
| author | Yu Chen Kai Huang Liang-Kuei Chen Liang-Kuei Chen Hui-Yu Wu Chih-Yu Hsu Yau-Sheng Tsai Wen-Chien Ko Pei-Jane Tsai Pei-Jane Tsai Pei-Jane Tsai |
| author_facet | Yu Chen Kai Huang Liang-Kuei Chen Liang-Kuei Chen Hui-Yu Wu Chih-Yu Hsu Yau-Sheng Tsai Wen-Chien Ko Pei-Jane Tsai Pei-Jane Tsai Pei-Jane Tsai |
| author_sort | Yu Chen |
| collection | DOAJ |
| description | Clostridium difficile, an obligate anaerobic gram-positive bacillus, generates spores and is commonly found colonizing the human gut. Patients with C. difficile infection (CDI) often exhibit clinical manifestations of pseudomembranous colitis or antibiotic-associated diarrhea. Surface layer proteins (SLPs) are the most abundant proteins in the C. difficile cell wall, suggesting that they might involve in immune recognition. Our previous results demonstrated that C. difficile triggers inflammasome activation. Here, we found SLPs as well as C. difficile induced inflammasome activation, and in a dose-dependent manner. In addition, the cholesterol-rich microdomains on the cell membrane (also referred to as lipid rafts) are thought to be crucial for bacterial adhesion and signal transduction. We demonstrated that lipid rafts participated in C. difficile SLPs binding to the cell membrane. Fluorescence microscopy showed that membrane cholesterol depletion by methyl-β-cyclodextrin (MβCD) reduced the association of SLPs with the cell surface. The coalescence of SLPs in the cholesterol-rich microdomains was confirmed in C. difficile-infected cells. Furthermore, the inflammasome activations induced by SLPs or C. difficile were abrogated by MβCD. Our results demonstrate that SLPs recruit the lipid rafts, which may be a key step for C. difficile colonization and inducing inflammasome activation. |
| first_indexed | 2024-12-10T22:46:46Z |
| format | Article |
| id | doaj.art-55884e9951ed497eb2bbbe96aff024f3 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-10T22:46:46Z |
| publishDate | 2020-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-55884e9951ed497eb2bbbe96aff024f32022-12-22T01:30:32ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-07-011110.3389/fimmu.2020.01675557483Membrane Cholesterol Is Crucial for Clostridium difficile Surface Layer Protein Binding and Triggering Inflammasome ActivationYu Chen0Kai Huang1Liang-Kuei Chen2Liang-Kuei Chen3Hui-Yu Wu4Chih-Yu Hsu5Yau-Sheng Tsai6Wen-Chien Ko7Pei-Jane Tsai8Pei-Jane Tsai9Pei-Jane Tsai10Department of Laboratory Medicine, Mackay Memorial Hospital, New Taipei, TaiwanDepartment of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, TaiwanDepartment of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, TaiwanDepartment of Pathology, National Cheng Kung University Hospital, Tainan, TaiwanDepartment of Laboratory Medicine, Mackay Memorial Hospital, New Taipei, TaiwanDepartment of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, TaiwanInstitute of Clinical Medicine, National Cheng Kung University, Tainan, TaiwanDepartment of Internal Medicine, National Cheng Kung University Hospital, Tainan, TaiwanDepartment of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, TaiwanDepartment of Pathology, National Cheng Kung University Hospital, Tainan, TaiwanCenter of Infectious Disease and Signaling Research, National Cheng Kung University, Tainan, TaiwanClostridium difficile, an obligate anaerobic gram-positive bacillus, generates spores and is commonly found colonizing the human gut. Patients with C. difficile infection (CDI) often exhibit clinical manifestations of pseudomembranous colitis or antibiotic-associated diarrhea. Surface layer proteins (SLPs) are the most abundant proteins in the C. difficile cell wall, suggesting that they might involve in immune recognition. Our previous results demonstrated that C. difficile triggers inflammasome activation. Here, we found SLPs as well as C. difficile induced inflammasome activation, and in a dose-dependent manner. In addition, the cholesterol-rich microdomains on the cell membrane (also referred to as lipid rafts) are thought to be crucial for bacterial adhesion and signal transduction. We demonstrated that lipid rafts participated in C. difficile SLPs binding to the cell membrane. Fluorescence microscopy showed that membrane cholesterol depletion by methyl-β-cyclodextrin (MβCD) reduced the association of SLPs with the cell surface. The coalescence of SLPs in the cholesterol-rich microdomains was confirmed in C. difficile-infected cells. Furthermore, the inflammasome activations induced by SLPs or C. difficile were abrogated by MβCD. Our results demonstrate that SLPs recruit the lipid rafts, which may be a key step for C. difficile colonization and inducing inflammasome activation.https://www.frontiersin.org/article/10.3389/fimmu.2020.01675/fullClostridium difficilemembrane cholesterollipid raftssurface layer proteinsinflammasome activation |
| spellingShingle | Yu Chen Kai Huang Liang-Kuei Chen Liang-Kuei Chen Hui-Yu Wu Chih-Yu Hsu Yau-Sheng Tsai Wen-Chien Ko Pei-Jane Tsai Pei-Jane Tsai Pei-Jane Tsai Membrane Cholesterol Is Crucial for Clostridium difficile Surface Layer Protein Binding and Triggering Inflammasome Activation Frontiers in Immunology Clostridium difficile membrane cholesterol lipid rafts surface layer proteins inflammasome activation |
| title | Membrane Cholesterol Is Crucial for Clostridium difficile Surface Layer Protein Binding and Triggering Inflammasome Activation |
| title_full | Membrane Cholesterol Is Crucial for Clostridium difficile Surface Layer Protein Binding and Triggering Inflammasome Activation |
| title_fullStr | Membrane Cholesterol Is Crucial for Clostridium difficile Surface Layer Protein Binding and Triggering Inflammasome Activation |
| title_full_unstemmed | Membrane Cholesterol Is Crucial for Clostridium difficile Surface Layer Protein Binding and Triggering Inflammasome Activation |
| title_short | Membrane Cholesterol Is Crucial for Clostridium difficile Surface Layer Protein Binding and Triggering Inflammasome Activation |
| title_sort | membrane cholesterol is crucial for clostridium difficile surface layer protein binding and triggering inflammasome activation |
| topic | Clostridium difficile membrane cholesterol lipid rafts surface layer proteins inflammasome activation |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2020.01675/full |
| work_keys_str_mv | AT yuchen membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT kaihuang membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT liangkueichen membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT liangkueichen membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT huiyuwu membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT chihyuhsu membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT yaushengtsai membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT wenchienko membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT peijanetsai membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT peijanetsai membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation AT peijanetsai membranecholesteroliscrucialforclostridiumdifficilesurfacelayerproteinbindingandtriggeringinflammasomeactivation |