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Three-Dimensional Human Bronchial-Tracheal Epithelial Tissue-Like Assemblies (TLAs) as Hosts for Severe Acute Respiratory Syndrome (SARS)-CoV InfectionA three-dimensional (3-D) tissue-like assembly (TLA) of human bronchial-tracheal mesenchymal (HBTC) cells with an overlay of human bronchial epithelial (BEAS-2B) cells was constructed using a NASA Bioreactor to survey the infectivity of SARS-CoV. This TLA was inoculated with a low passage number Urbani strain of SARS-CoV. At selected intervals over a 10-day period, media and cell aliquots of the 3-D TLA were harvested for viral titer assay and for light and electron microscopy examination. All viral titer assays were negative in both BEAS-2B two-dimensional monolayer and TLA. Light microscopy immunohistochemistry demonstrated antigen-antibody reactivity with anti-SARS-CoV polyclonal antibody to spike and nuclear proteins on cell membranes and cytoplasm. Coronavirus Group 2 cross-reactivity was demonstrated by positive reaction to anti-FIPV 1 and anti-FIPV 1 and 2 antibodies. TLA examination by transmission electron microscopy indicated increasing cytoplasmic vacuolation with numerous electron-dense bodies measuring 45 to 270 nm from days 4 through 10. There was no evidence of membrane blebbing, membrane duplication, or fragmentation of organelles in the TLAs. However, progressive disruption of endoplasmic reticulum was observed throughout the cells. Antibody response to SARS-CoV specific spike and nucleocapsid glycoproteins, cross-reactivity with FIPV antibodies, and the cytoplasmic pathology suggests this HBTE TLA model is permissive to SARS-CoV infection.
Document ID
20070002509
Document Type
Technical Publication (TP)
Authors
Suderman, M. T. (NASA Johnson Space Center Houston, TX, United States)
McCarthy, M. (NASA Johnson Space Center Houston, TX, United States)
Mossell, E. (Texas Univ. Galveston, TX, United States)
Watts, D. M. (Texas Univ. Galveston, TX, United States)
Peters, C. J. (Texas Univ. Galveston, TX, United States)
Shope, R. (Texas Univ. Galveston, TX, United States)
Goodwin, T. J. (NASA Johnson Space Center Houston, TX, United States)
Date Acquired
August 24, 2013
Publication Date
June 1, 2006
Subject Category
Life Sciences (General)
Report/Patent Number
S-980
JSC-CN-10639
NASA/TP-2006-213723
Funding Number(s)
CONTRACT_GRANT: 279-00-0308
Distribution Limits
Public
Copyright
Public Use Permitted.

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