Paramyxovirus Infection Mimics In Vivo Cellular Dynamics in Three-Dimensional Human Bronchio-Epithelial Tissue-Like Assemblies
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چکیده
....................................................................................................................................................... 1 1.0 Results ................................................................................................................................................... 3 Characterization of 3D HBE TLAs .......................................................................................................... 3 Expression of HRE-specific markers ........................................................................................................ 5 Gene expression in BEAS-2B monolayers and 3D HBE TLAs ................................................................... 7 wtRSV and wtPIV3 replicate more efficiently than mutants in TLAs ...................................................... 8 Innate immune response to RSV and PIV3 infection .............................................................................. 9 TLAs predict in vivo inflammatory responses ..................................................................................... 10 2.0 Discussion ............................................................................................................................................ 14 3.0 Methods ............................................................................................................................................... 19 Cells ....................................................................................................................................................... 19 3D TLA cultures ..................................................................................................................................... 19 Viruses .................................................................................................................................................. 19 Scanning Electron Microscopy of 3D HBE TLAs .................................................................................... 20 Transmission Electron Microscopy of 3D HBE TLAs .............................................................................. 20 Expression of epithelial markers by confocal microscopy .................................................................... 21 Expression of epithelial markers by immunohistochemistry ................................................................ 22 Host transcriptional profiling data analysis ......................................................................................... 22 In vitro virus inoculation and titration ................................................................................................ 24 Human nasal wash samples ................................................................................................................ 24 Quantifying cytokines .......................................................................................................................... 25 4.0 Supplementary Methods .................................................................................................................... 26 Construction of rbPIV3 Shipping Fever ............................................................................................... 26 5.0 Acknowledgments ................................................................................................................................ 28 6.0 References ............................................................................................................................................ 29
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تاریخ انتشار 2012