期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 117, 期 3, 页码 1700-1710出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1910138117
关键词
norovirus; enteroid/organoid; bile acid; acid sphingomyelinase; ceramide
资金
- NIH [P01 AI57788, R01AG034389, R01NS095215, CA125123]
- PHS Grants from the NIH [P01AI 057788, P30 DK 56338]
- Agriculture and Food Research Initiative Competitive Grant from the US Department of Agriculture, National Institute of Food and Agriculture [2011-6800330395]
- Japan Agency for Medical Research and Development [JP18fk0108034, JP19fk0108102]
- Japan Society for the Promotion of Science KAKENHI Grant [JP18K07153]
- National Science Foundation [1615874]
- Advanced Technology Core Laboratories (Baylor College of Medicine)
- CPRIT [RP150578, RP170719]
- NIH S10 Grant [1S10OD023469]
- Direct For Biological Sciences
- Div Of Molecular and Cellular Bioscience [1615874] Funding Source: National Science Foundation
Human noroviruses (HuNoVs) cause sporadic and epidemic outbreaks of gastroenteritis in all age groups worldwide. We previously reported that stem cell-derived human intestinal enteroid (HIE) cultures support replication of multiple HuNoV strains and that some strains (e.g., GII.3) replicate only in the presence of bile. Heat-and trypsin-treatment of bile did not reduce GII.3 replication, indicating a nonproteinaceous component in bile functions as an active factor. Here we show that bile acids (BAs) are critical for GII.3 replication and replication correlates with BA hydrophobicity. Using the highly effective BA, glycochenodeoxycholic acid (GCDCA), we show BAs act during the early stage of infection, BA-dependent replication in HIEs is not mediated by detergent effects or classic farnesoid X receptor or Takeda G protein-coupled receptor 5 signaling but involves another G protein-coupled receptor, sphingosine-1-phosphate receptor 2, and BA treatment of HIEs increases particle uptake. We also demonstrate that GCDCA induces multiple cellular responses that promote GII.3 replication in HIEs, including enhancement of 1) endosomal uptake, 2) endosomal acidification and subsequent activity of endosomal/lysosomal enzyme acid sphingomyelinase (ASM), and 3) ceramide levels on the apical membrane. Inhibitors of endosomal acidification or ASM reduce GII.3 infection and exogenous addition of ceramide alone permits infection. Furthermore, inhibition of lysosomal exocytosis of ASM, which is required for ceramide production at the apical surface, decreases GII.3 infection. Together, our results support a model where GII.3 exploits rapid BA-mediated cellular endolysosomal dynamic changes and cellular ceramide to enter and replicate in jejunal HIEs.
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